Adenosine triphosphate(ATP)induced cell death(AICD)is a critical cellular process that has garnered substantial scientific interest for its profound relevance to cancer biology and to therapeutic interventions.This co...Adenosine triphosphate(ATP)induced cell death(AICD)is a critical cellular process that has garnered substantial scientific interest for its profound relevance to cancer biology and to therapeutic interventions.This comprehensive review unveils the intricate web of AICD mechanisms and their intricate connections with cancer biology.This review offers a comprehensive framework for comprehending the multifaceted role of AICD in the context of cancer.This is achieved by elucidating the dynamic interplay between systemic and cellular ATP homeostasis,deciphering the intricate mechanisms governing AICD,elucidating its intricate involvement in cancer signaling pathways,and scrutinizing validated key genes.Moreover,the exploration of AICD as a potential avenue for cancer treatment underscores its essential role in shaping the future landscape of cancer therapeutics.展开更多
After hypoxia, ischemia, or inflammatory injuries to the central nervous system, the damaged cells release a large amount of adenosine triphosphate, which may cause secondary neuronal death. Autophagy is a form of cel...After hypoxia, ischemia, or inflammatory injuries to the central nervous system, the damaged cells release a large amount of adenosine triphosphate, which may cause secondary neuronal death. Autophagy is a form of cell death that also has neuroprotective effects. Cell Counting Kit assay, monodansylcadaverine staining, flow cytometry, western blotting, and real-time PCR were used to determine the effects of exogenous adenosine triphosphate treatment at different concentrations (2, 4, 6, 8, 10 mmol/L) over time (1, 2, 3, and 6 hours) on the apoptosis and autophagy of SH-SY5Y cells. High concentrations of extracellular adenosine triphosphate induced autophagy and apoptosis of SH-SYSY cells. The enhanced autophagy first appeared, and peaked at 1 hour after treatment with adenosine triphosphate. Cell apoptosis peaked at 3 hours, and persisted through 6 hours. With prolonged exposure to the adenosine triphosphate treatment, the fraction of apoptotic cells increased. These data suggest that the SH-SY5Y neural cells initiated autophagy against apoptosis within an hour of adenosine triphosphate treatment to protect themselves against injury.展开更多
The mammalian target of rapamycin (mTOR) pathway plays an important role in neuronal growth, proliferation and differentiation. To better understand the role of mTOR pathway involved in the induction of spinal cord ...The mammalian target of rapamycin (mTOR) pathway plays an important role in neuronal growth, proliferation and differentiation. To better understand the role of mTOR pathway involved in the induction of spinal cord injury, rat models of spinal cord injury were established by modified Allen's stall method and interfered for 7 days by intraperitoneal administration of mTOR activator adenosine triphosphate and mTOR kinase inhibitor rapamycin. At 1-4 weeks after spinal cord injury induction, the Basso, Beattie and Bresnahan locomotor rating scale was used to evaluate rat locomotor function, and immunohistochemical staining and western blot analysis were used to detect the expression of nestin (neural stem cell marker), neuronal nuclei (neuronal marker), neuron specific enolase, neurofilament protein 200 (axonal marker), glial fibrillary acidic protein (astrocyte marker), Akt, mTOR and signal transduction and activator of transcription 3 (STAT3). Results showed that adenosine triphosphate-mediated Akt/mTOR/STAT3 pathway increased endogenous neural stem cells, induced neurogenesis and axonal growth, inhibited excessive astrogliosis and improved the locomotor function of rats with spinal cord injury.展开更多
In this study,we aimed at developing an efficient biocatalytic process for bio-production of cyclic adenosine monophosphate(c AMP)from adenosine triphosphate(ATP).First,adenylate cyclase from Escherichia coli MG1655(E...In this study,we aimed at developing an efficient biocatalytic process for bio-production of cyclic adenosine monophosphate(c AMP)from adenosine triphosphate(ATP).First,adenylate cyclase from Escherichia coli MG1655(EAC)and Bordetella Pertussis(BAC)were expressed in E.coli BL21(DE3)and comparatively analyzed for their activities.As a result,EAC from E.coli MG1655 exhibited a higher activity.However,amount of EAC were obtained in an insoluble form.Therefore,we expressed the first 446 amino acids of EAC(EAC446)to avoid the inclusion body.The effects of induction temperature,incubation time,and incubation p H were further evaluated to improve the expression of EAC446.Subsequently,the reaction process for the production of c AMP with ATP as a starting material was investigated.As none of c AMP was detected in the whole-cell based biocatalytic process,the reaction catalyzed by the crude enzyme was determined for c AMP production.What's more,the reaction temperature,reaction p H,metal ion additives and substrate concentration was optimized,and the maximum c AMP production of 18.45 g·L^-1was achieved with a yield of 95.4%after bioconversion of 6 h.展开更多
Nicotinamide adenine dinucleotide (NAD) oscillation was observed when the isolated mitochondria were immersed in a pyruvate solution. In addition, when an adenosine diphosphate (ADP) was added to the mitochondrial sus...Nicotinamide adenine dinucleotide (NAD) oscillation was observed when the isolated mitochondria were immersed in a pyruvate solution. In addition, when an adenosine diphosphate (ADP) was added to the mitochondrial suspension containing pyruvate, adenosine triphosphate (ATP) oscillation was observed as well as NADH oscillation. At this time, the pH within mitochondria also oscillated. It was found that the oscillatory reaction of NADH caused by the membrane permeation of pyruvate continues, causing the oscillation of NADH and H+ in the subsequent reactions. The pH oscillation led to the ATP oscillation. It is considered that the oscillatory reaction caused by the gradual entry of pyruvate into mitochondria was thought to be carried over to both the citric acid cycle and the respiratory chain, ultimately leading to the ATP oscillation in oxidative phosphorylation. Similarly, it was found that membrane permeation of malate causes the gradual occurrence of NADH, at which point NADH oscillates, followed by an oscillatory reaction of the respiratory chain, and finally ATP oscillation. It was found that the oscillations of NADH and ATP occur without going through the citric acid cycle. Oscillations of NADH and other intermediates in both the citric acid cycle and respiratory chain were also confirmed by experiments using semipermeable membranes. These results support our hypothesis that the gradual entry of the substrate by membrane permeation triggers an oscillatory reaction of the enzyme, which is also carried over to subsequent reactions.展开更多
Background Repetitive transcranial magnetic stimulation (rTMS) research has mainly been focused on the therapeutic effect of psychiatric disorders and Parkinson's disease. A few studies have shown that rTMS might p...Background Repetitive transcranial magnetic stimulation (rTMS) research has mainly been focused on the therapeutic effect of psychiatric disorders and Parkinson's disease. A few studies have shown that rTMS might protect against delayed neuronal death induced by transient ischemia, enhance long-term potentiation in ischemic conditions and affect regional brain blood flow and metabolism. The aim of this study was to determine the effects of repetitive transcranial magnetic stimulation (rTMS) on adenosine triphosphate (ATP) content and microtubule associated protein-2 (MAP-2) expression in rat brain after middle cerebral artery occlusion (MCAO)/reperfusion. Methods To study the effects of different timecourses of rTMS on ATP content and MAP-2 expression, 90 rats were randomly divided into three groups (30 rats in each group). To study the effects of multiple rTMS parameters on ATP content and MAP-2 expression, the rats in each group were further divided into six subgroups (five rats each). The rats were sacrificed at 1-hour, 24-hour and 48-hour intervals after reperfusion, and the brain tissues were collected for the detection of ATP and MAP-2. Results rTMS could significantly increase ATP content and MAP-2 expression in the left brain following ischemic insult (P 〈0.01) and different rTMS parameters had different effects on the ATP level and the MAP-2 expression in the left striatum. A high-frequency rTMS played an important role in MAP-2 expression and ATP preservation. Conclusions This study revealed that rTMS induced significant increase of ATP content and MAP-2 expression in the injured area of the brain, suggesting that the regulation of both ATP and MAP-2 may be involved in the biological mechanism of the effect of rTMS on neural recovery. Therefore, rTMS may become a potential adjunctive therapy for ischemic cerebrovascular disease.展开更多
In this work,polymethacrylic acid(PMAA)-templated silver nanoclusters(Ag NCs)were developed as the fluorescent probe for the efficient and sensitive detection of adenosine triphosphate(ATP)in a wide range of pH values...In this work,polymethacrylic acid(PMAA)-templated silver nanoclusters(Ag NCs)were developed as the fluorescent probe for the efficient and sensitive detection of adenosine triphosphate(ATP)in a wide range of pH values.The fluorescence intensity of the Ag NCs could keep stable with pH values ranging from2.5 to 9.3.The detection of ATP was based on the quenching of the fluorescent Ag NCs in the presence of ATP.The fluorescence quenching of the Ag NCs with increasing ATP concentration was studied at pH 2.5,4.5,7.0 and 8.5 which involved a wide pH environment in body fluids.The limit of detection(LOD)for ATP was as low as 0.1 mmol/L in an acidic environment with pH of 2.5 and all the linear correlation coefficients were satisfactory under wide-span pH values from 2.5 to 8.5.In addition,the sensitive determination of ATP was also achieved by adding copper ions(Cu^2+).The high selectivity and rapid detection process proved that the fluorescent probe had great potential to detect ATP in biological samples under different pH conditions.展开更多
Transmembrane transport plays an important role in many physiological functions,and mimicking this biological process in artificial systems has potential applications in biosensing,drug delivery,and bionic science.Her...Transmembrane transport plays an important role in many physiological functions,and mimicking this biological process in artificial systems has potential applications in biosensing,drug delivery,and bionic science.Here,a lipophilic split aptamer was developed as a novel transmembrane carrier for adenosine triphosphate(ATP)transport.The ATP carrier comprises two split aptamer fragments and cholesterol tags,with the split aptamers acting as targetrecognition domains to enhance their specific binding capability and the cholesterol tags as hydrophobic domains to facilitate membrane penetration.Giant unilamellar vesicle experiments demonstrated that the ATP carrier-mediated transmembrane transport was concentration-and time-dependent and showed high transport selectivity.Moreover,the artificial carriers were applicable to living cells and facilitated rapid cell internalization of fluorescencelabeled ATP.Furthermore,carrier-mediated ATP transport into ATP-deficient cells enabled recovery of cellular ATP levels and improved cell viability.This study demonstrated the efficacy of an aptamer nanostructure for designing DNA-based synthetic carriers with high selectivity and flexibility.展开更多
The activation of adenosine A1 receptors is important for protecting against ischemic brain injury and pretreatment with electroacupuncture has been shown to mitigate ischemic brain insult. The aim of this study was t...The activation of adenosine A1 receptors is important for protecting against ischemic brain injury and pretreatment with electroacupuncture has been shown to mitigate ischemic brain insult. The aim of this study was to test whether the adenosine A1 receptor mediates electroacupuncture pretreatment-induced neuroprotection against ischemic brain injury. We first performed 30 minutes of electroacupuncture pretreatment at the Baihui acupoint(GV20), delivered with a current of 1 mA, a frequency of 2/15 Hz, and a depth of 1 mm. High-performance liquid chromatography found that adenosine triphosphate and adenosine levels peaked in the cerebral cortex at 15 minutes and 120 minutes after electroacupuncture pretreatment, respectively. We further examined the effect of 15 or 120 minutes electroacupuncture treatment on ischemic brain injury in a rat middle cerebral artery-occlusion model. We found that at 24 hours reperfusion,120 minutes after electroacupuncture pretreatment, but not for 15 minutes, significantly reduced behavioral deficits and infarct volumes. Last, we demonstrated that the protective effect gained by 120 minutes after electroacupuncture treatment before ischemic injury was abolished by pretreatment with the A1-receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine(1 mg/kg, intraperitoneally). Our results suggest that pretreatment with electroacupuncture at the Baihui acupoint elicits protection against transient cerebral ischemia via action at adenosine A1 receptors.展开更多
Traumatic spinal cord injuries interrupt the connection of all axonal projections with their neuronal targets below and above the lesion site. This interruption results in either temporary or permanent alterations in ...Traumatic spinal cord injuries interrupt the connection of all axonal projections with their neuronal targets below and above the lesion site. This interruption results in either temporary or permanent alterations in the locomotor, sensory, and autonomic functions. Damage in the spinal tissue prevents the re-growth of severed axons across the lesion and their reconnection with neuronal targets. Therefore, the absence of spontaneous repair leads to sustained impairment in voluntary control of movement below the injury. For decades, axonal regeneration and reconnection have been considered the opitome of spinal cord injury repair with the goal being the repair of the damaged long motor and sensory tracts in a complex process that involves:(1) resealing injured axons;(2) reconstructing the cytoskeletal structure inside axons;(3) re-establishing healthy growth cones;and(4) assembling axonal cargos. These biological processes require an efficient production of adenosine triphosphate, which is affected by mitochondrial dysfunction after spinal cord injury. From a pathological standpoint, during the secondary stage of spinal cord injury, mitochondrial homeostasis is disrupted, mainly in the distal segments of severed axons. This result in a reduction of adenosine triphosphate levels and subsequent inactivation of adenosine triphosphate-dependent ion pumps required for the regulation of ion concentrations and reuptake of neurotransmitters, such as glutamate. The consequences are calcium overload, reactive oxygen species formation, and excitotoxicity. These events are intimately related to the activation of necrotic and apoptotic cell death programs, and further exacerbate the secondary stage of the injury, being a hallmark of spinal cord injury. This is why restoring mitochondrial function during the early stage of secondary injury could represent a potentially effective therapeutic intervention to overcome the motor and sensory failure produced by spinal cord injury. This review discusses the most recent evidence linking mitochondrial dysfunction with axonal regeneration failure in the context of spinal cord injury. It also covers the future of mitochondria-targeted therapeutical approaches, such as antioxidant molecules, removing mitochondrial anchor proteins, and increasing energetic metabolism through creatine treatment. These approaches are intended to enhance functional recovery by promoting axonal regenerationreconnection after spinal cord injury.展开更多
In our previous study, we found that the edible alcohol extract of the root of the medicinal plant Rhodiola crenulata(RCE) improved spatial cognition in a rat model of Alzheimer's disease. Another study from our la...In our previous study, we found that the edible alcohol extract of the root of the medicinal plant Rhodiola crenulata(RCE) improved spatial cognition in a rat model of Alzheimer's disease. Another study from our laboratory showed that RCE enhanced neural cell proliferation in the dentate gyrus of the hippocampus and prevented damage to hippocampal neurons in a rat model of chronic stress-induced depression. However, the mechanisms underlying the neuroprotective effects of RCE are unclear. In the present study, we investigated the anti-apoptotic effect of RCE and its neuroprotective mechanism of action in a rat model of Alzheimer's disease established by intracerebroventricular injection of streptozotocin. The rats were pre-administered RCE at doses of 1.5, 3.0 or 6.0 g/kg for 21 days before model establishment. ATP and cytochrome c oxidase levels were significantly decreased in rats with Alzheimer's disease. Furthermore, neuronal injury was obvious in the hippocampus, with the presence of a large number of apoptotic neurons. In comparison, in rats given RCE pretreatment, ATP and cytochrome c oxidase levels were markedly increased, the number of apoptotic neurons was reduced, and mitochondrial injury was mitigated. The 3.0 g/kg dose of RCE had the optimal effect. These findings suggest that pretreatment with RCE prevents mitochondrial dysfunction and protects hippocampal neurons from apoptosis in rats with Alzheimer's disease.展开更多
Aim: To investigate the possible role of manganese in the regulation of mitochondrial aconitase (mACON) activity human prostate carcinoma cell line PC-3 cells. Methods: The mACON enzymatic activities of human pros...Aim: To investigate the possible role of manganese in the regulation of mitochondrial aconitase (mACON) activity human prostate carcinoma cell line PC-3 cells. Methods: The mACON enzymatic activities of human prostate carcinoma cell line PC-3 cells were determined using a reduced nicotinamide adenine dinucleotide-coupled assay. Immunoblot and transient gene expression assays were used to study gene expression of the mACON. The putative response element for gene expression was identified using reporter assays with site-directed mutagenesis and electrophoretic mobility-shift assays. Results: In vitro study revealed that manganese chloride (MnCI2) treatment for 16 h inhibited the enzymatic activity of mACON, which induced the inhibition of citrate utility and cell proliferation of PC- 3 cells. Although results from transient gene expression assays showed that MnCI2 treatment upregulated gene translation by approximately 5-fold through the iron response element pathway, immunoblot and reporter assays showed that MnCl2 treatments inhibited protein and gene expression of mACON. This effect was reversed by cotreatment with ferric ammonium citrate. Additional reporter assays with site-directed mutagenesis and electrophoretic mobility-shift assays suggested that a putative metal response element in the promoter of the mACON gene was involved in the regulation of MnCh on the gene expression of mACON. Conclusion: These findings suggest that manganese acts as an antagonist of iron, disrupting the enzymatic activity and gene expression of mACON and citrate metabolism in the prostate.展开更多
Summary: Relationship between ATP changes of rabbit blood and postmortem interval (PMI) was studied. Twenty-four healthy rabbits were sacrificed and randomly divided into 3 groups with 8 rab- bits of each group. Th...Summary: Relationship between ATP changes of rabbit blood and postmortem interval (PMI) was studied. Twenty-four healthy rabbits were sacrificed and randomly divided into 3 groups with 8 rab- bits of each group. The bodies of three groups were placed in calorstat at temperature of 15℃, 25℃ and 35℃, respectively. The blood from the right ventricle was sampled through indwelling needle each 4 h until 72 h after death. ATP levels in the blood samples were measured by using ATP fluo- rescence rapid detection technique at different PMIs. Blood ATP levels slightly increased in the early stage after death and then constantly declined at all temperatures (15℃, 25℃, and 35℃). Cubic polynomial regression equations with log[ATP] as dependent variable (y) and PMI as independent variable (x) at different temperatures and the optimal time period were established as followed: Under 15℃ and during 16-64 h after death, y=-3.027×10^-5x^3+0.003x^2-0.096x-10.625 (Ra^2=0.992, P〈0.001); under 25℃ and during 8-56 h after death, y=-2.921×10^-5x^3+0.002x^2- 0.059x-11.186 (Ra^2=0.989, P〈0.001); under 35℃ and during 4-36 h after death, y=-9.769×10^-5x^3+ 0.005x^2 -0.117x-11.166 (Ra^2=0.991, P〈0.001). The changes in ATP levels in blood collected from right ven- tricle of rabbit cadavers showed relatively stable and regular degradation within 72 h after death at different temperatures.展开更多
AIM: To investigate the potential of pigment epitheliumderived factor(PEDF) to protect the immortalized rat retinal ganglion cells-5(RGC-5) exposed to Co Cl2-induced chemical hypoxia. METHODS: After being differ...AIM: To investigate the potential of pigment epitheliumderived factor(PEDF) to protect the immortalized rat retinal ganglion cells-5(RGC-5) exposed to Co Cl2-induced chemical hypoxia. METHODS: After being differentiated with staurosporine(SS), RGC-5 cells were cultured in four conditions: control group cells cultured in Dulbecco 's modified eagle medium(DMEM) supplemented with 10% fetal bovine serum, 100 μmol/m L streptomycin and penicillin(named as normal conditions); hypoxia group cells cultured in DMEM containing 300 μmol/m L Co Cl2; cells in the group protected by PEDF were first pretreated with 100 ng/m L PEDF for 2h and then cultured in the same condition as hypoxia group cells; and PEDF group cells that were cultured in the presence of 100 ng/m L PEDF under normal conditions. The cell viability was assessed by MTT assay, the percentage of apoptotic cells was quantified using Annexin V-FITC apoptosis kit, and intra-cellar reactive oxygen species(ROS) was measured by dichloro-dihydro-fluorescein diacetate(DCFH-DA) probe. The mitochondria-mediated apoptosis was also examined to further study the underlying mechanism of the protective effect of PEDF. The opening of mitochondrial permeability transition pores(m PTPs) and membrane potential(Δψm) were tested as cellular adenosine triphosphate(ATP) level and glutathione(GSH). Also, the expression and distribution of Cyt C and apoptosis inducing factor(AIF) were observed.RESULTS: SS induced differentiation of RGC-5 cells resulting in elongation of their neurites and establishing contacts between outgrowths. Exposure to 300 μmol/m L Co Cl2 triggered death of 30% of the total cells in cultures within 24 h. At the same time, pretreatment with 100 ng/m L PEDF significantly suppressed the cell death induced by hypoxia(P〈0.05). The apoptosis induced by treatment of Co Cl2 was that induced cell death accompanied with increasing intracellar ROS and decreasing GSH and ATP level. PEDF pretreatment suppressed these effects(P〈0.05). Additionally, PEDF treatment inhibited the opening of m PTPs and suppressed decreasing of Δψm in RGC-5 cells, resulting in blocking of the mitochondrial apoptotic pathway.CONCLUSION: Pretreatment of RGC-5 cells with 100 ng/m L PEDF significantly decreases the extent of apoptosis. PEDF inhibits the opening of m PTPs and suppresses decreasing of Δψm. Moreover, PEDF also reduces ROS production and inhibits cellular ATP level's reduction. Cyt C and AIF activation in PEDF-pretreated cultures are also reduced. These results demonstrate the potential for PEDF to protect RGCs against hypoxic damage in vitro by preventing mitochondrial dysfunction.展开更多
AIM:To measure the expression of transient receptor potential cation channel subfamily V member 4(TRPV4)in the rat cornea and determine whether it is related to adenosine triphosphate(ATP)generation in a rat model of ...AIM:To measure the expression of transient receptor potential cation channel subfamily V member 4(TRPV4)in the rat cornea and determine whether it is related to adenosine triphosphate(ATP)generation in a rat model of acute ocular hypertension(AOH).METHODS:Immunofluorescence staining of TRPV4,P2X2 receptor,P2X3 receptor,and(33-tubulin in rat corneal longitudinal sections and paved was performed to clearly display histological structures.Rat models of AOH and agonist/antagonist-treated groups were established and corneal ATP was measured using an ATP assay.The independent t-test and simple linear correlation model were adopted for statistical analyses.RESULTS:Immunofluorescence staining of rat cornea sections revealed that epithelial and endothelial membranes showed strong immunoreactivity for TRPV4 and P2X2 receptor and coexpression with(33-tubulin in the rat corneal epithelial layer.Corneal ATP was significantly higher in the AOH rat model than in the control(P<0.05)and apparently lower after pretreatment by applying eyedrops of TRPV4 antagonist RN1734 with 30-40 mm Hg intraocular pressure(IOP;P<0.05).A simple linear regression model showed a positive correlation between rat corneal ATP and IOP values(R^(2)=0.996,P=0.0134)from the normal IOP(113 mm Hg)to 40 mm Hg.At 10-40min after anterior chamber injection of GSK1016790A(0.01 mL,50 nmol/L in 0.9%NaCl),corneal ATP was significantly higher than in the control group(P<0.05),which peaked at 1Omin.The ATP concentration of the normal epithelium was higher than that of the endothelium in the AOH rat model and after anterior chamber injection of GSK1016790A(P<0.05).CONCLUSION:The ATP concentration in the AOH rat cornea is increased by TRPV4 activation.展开更多
This study investigated the protective effect of ATP on skeletal muscle satellite cells damaged by H2O2 in neonatal rats and the possible mechanism. The skeletal muscle satellite cells were randomly divided into four ...This study investigated the protective effect of ATP on skeletal muscle satellite cells damaged by H2O2 in neonatal rats and the possible mechanism. The skeletal muscle satellite cells were randomly divided into four groups: normal group, model group(cells treated with 0.1 mmol/L H2O2 for 50 s), protection group(cells treated with 16, 8, 4, 2, 1, 0.5, or 0.25 mmol/L ATP for 24 h, and then with 0.1 mmol/L H2O2 for 50 s), proliferation group(cells treated with 16, 8, 4, 2, 1, 0.5, or 0.25 mmol/L ATP for 24 h). MTT assay, FITC+PI+DAPI fluorescent staining, Giemsa staining and immunofluorescence were performed to examine cell viability and apoptosis, and apoptosis-related proteins. The results showed that the survival rate of skeletal muscle satellite cells was decreased and the apoptosis rate was increased after H2O2 treatment(P〈0.01). Different doses of ATP had different effects on skeletal muscle satellite cells damaged by H2O2: the survival rate of muscle satellite cells treated with ATP at 4, 2, or 1 mmol/L was increased. The protective effect was most profound on cells treated with 2 mmol/L ATP. Immunofluorescence showed that ATP could increase the number of Bcl-2-positive cells(P〈0.01) and decrease the number of the Bax-positive cells(P〈0.01). It was concluded that ATP could protect skeletal muscle satellite cells against H2O2 damage in neonatal rats, which may be attributed to the up-regulation of the expression of Bcl-2 and down-regulation of Bax, resulting in the suppression of apoptosis.展开更多
In consideration of the mechanism for shear-stress-induced Ca^2+ influx via ATP(adenosine triphosphate)-gated ion channel P2X4 in vascular endothelial cells, a modified model is proposed to describe the shear-stres...In consideration of the mechanism for shear-stress-induced Ca^2+ influx via ATP(adenosine triphosphate)-gated ion channel P2X4 in vascular endothelial cells, a modified model is proposed to describe the shear-stress-induced Ca^2+ influx. It is affected both by the Ca^2+ gradient across the cell membrane and extracellular ATP concentration on the cell surface. Meanwhile, a new static ATP release model is constructed by using published experimental data. Combining the modified intracellular calcium dynamics model with the new ATP release model, we establish a nonlinear Ca^2+ dynamic system in vascular endothelial cells. The ATP-mediated calcium response in vascular endothelial cells subjected to shear stresses is analyzed by solving the governing equations of the integrated dynamic system. Numerical results show that the shear-stress-induced calcium response predicted by the proposed model is more consistent with the experimental observations than that predicted by existing models.展开更多
The paper introduces a new type instrument for rapid measuring ATP.The system consists of a micromodule ATP sensor and an instrument for measuring weak light transmitted by optic fiber. The micromodule ATP sensor main...The paper introduces a new type instrument for rapid measuring ATP.The system consists of a micromodule ATP sensor and an instrument for measuring weak light transmitted by optic fiber. The micromodule ATP sensor mainly is composed of enzyme membrane, a probe and a bundle of optic fiber. The instrument measuring weak light consists of photomultiplier, high voltage power, pulse amplifier and counter. The instrument was characterized by simple structure,small size, rapid response time (<5 s), high sensitivity (10-12 mol/L), stable performance (measuring the same sample for 50 times, CV< 5% ), long enzyme storage time (>3 months).展开更多
BACKGROUND Adrenoleukodystrophy(ALD)is caused by a deficit in the ABCD1 gene,which leads to demyelination of neurons and dysfunction of the adrenal cortices and testicles.Of the three known phenotypes,30%-50%of male A...BACKGROUND Adrenoleukodystrophy(ALD)is caused by a deficit in the ABCD1 gene,which leads to demyelination of neurons and dysfunction of the adrenal cortices and testicles.Of the three known phenotypes,30%-50%of male ALD patients present with the adrenomyeloneuropathy phenotype,characterized by gait disturbance as the initial symptom.CASE SUMMARY A 46-year-old man with a prior diagnosis of ALD was admitted to a Korean medicine hospital for the treatment of gait disturbance.His ability to walk was severely impaired at admission,significantly affecting the patient’s quality of life.He was treated with acupuncture,pharmacopuncture,electroacupuncture,and herbal medicine for 23 d.The 25-Foot Walk test(25FW),timed up and go(TUG),comfortable gait speed(CGS),numeric rating scale(NRS),Berg Balance Scale(BBS),Tinetti test,manual muscle test(MMT),and 3-level version of EuroQol-5 dimension(EQ-5D-3L)were used to evaluate the patient.The outcomes of the 25FW,TUG,and CGS improved during hospitalization.From the time of admission to discharge we observed:A decrease in NRS scores in the lower extremities and the lower back;an increase of 3 points in the BBS;a 1-point increase in the balancing part of the Tinetti Test;MMT and EQ-5D-3L performances remained unchanged.CONCLUSION Traditional Chinese medicine treatments could be a therapeutic option to alleviate issues related to gait disturbance in ALD.展开更多
[Objectives]To detect the contents of components in Shuanghuanglian freeze-dried powder and to explore the effects of Shuanghuanglian freeze-dried powder and its components on human hepatocytes(HL-7702)alone or in com...[Objectives]To detect the contents of components in Shuanghuanglian freeze-dried powder and to explore the effects of Shuanghuanglian freeze-dried powder and its components on human hepatocytes(HL-7702)alone or in combination with cefradine.[Methods]High performance liquid chromatography(HPLC)was used to detect the contents of baicalin,wogonin,chlorogenic acid and forsythin,the main components of Shuanghuanglian freeze-dried powder.HL-7702 cells were cultured with Shuanghuanglian freeze-dried powder and the main components of Shuanghuanglian freeze-dried powder alone or in combination with cefradine.Enzyme linked immunosorbent assay(ELISA)was used to detect the contents of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in the cell supernatant after culture,and HPLC was used to detect the expression level of adenosine diphosphate(ADP)and adenosine triphosphate(ATP);agarose gel electrophoresis was used to detect the expression of cyclooxygenase 2(COX-2)and heme oxygenase-1(HO-1)in HL-7702 cells.[Results]In Shuanghuanglian freeze-dried powder for injection,the content of baicalin was the highest,and the content of wogonin was the lowest.Compared with the control group,the expressions of AST and ALT in human hepatocytes(HL-7702)in high-dose baicalin group,forsythin group and wogonin group decreased(P<0.05,P<0.01),while the expression of ALT in chlorogenic acid+cefradine group(0.046 mg/mL)and forsythin+cefradine group(0.046 mg/mL)increased(P<0.05,P<0.01),and the expression of AST had no significant difference(P>0.05);the results in the low-dose group were similar to those in the high-dose group.Compared with the control group,ATP expression in chlorogenic acid group,chlorogenic acid+cefradine group(0.046 mg/mL)and forsythin+cefradine group(0.046 mg/mL)in the high-dose group decreased(P<0.05,P<0.01),and ADP expression was not significantly different(P>0.05);in the low-dose group,the expression of ATP and ADP increased in baicalin group(P<0.05),but decreased in wogonin group,baicalin+cefradine group(0.046 mg/mL)and wogonin group+cefradine group(0.046 mg/mL)(P<0.05,P<0.01).Compared with the control group,the expressions of COX-2 and HO-1 in HL-7702 cells in the cefradine group showed no significant difference(P>0.05).The expression of HO-1 and COX-2 in the different dose groups of Shuanghuanglian and the group combined with cefradine increased,and the difference was significant(P<0.05,P<0.01).[Conclusions]The components of Shuanghuanglian freeze-dried powder for injection had effects on hepatocytes,of which baicalin had a significant effect,and the effect of cefradine on hepatocytes was increased when used in combination with cefradine.展开更多
基金Supported by National Natural Science Foundation of China,No.81960877University Innovation Fund of Gansu Province,No.2021A-076+4 种基金Gansu Province Science and Technology Plan(Innovation Base and Talent Plan),No.21JR7RA561Natural Science Foundation of Gansu Province,No.21JR1RA267 and No.22JR5RA582Education Technology Innovation Project of Gansu Province,No.2022A-067Innovation Fund of Higher Education of Gansu Province,No.2023A-088Gansu Province Science and Technology Plan International Cooperation Field Project,No.23YFWA0005.
文摘Adenosine triphosphate(ATP)induced cell death(AICD)is a critical cellular process that has garnered substantial scientific interest for its profound relevance to cancer biology and to therapeutic interventions.This comprehensive review unveils the intricate web of AICD mechanisms and their intricate connections with cancer biology.This review offers a comprehensive framework for comprehending the multifaceted role of AICD in the context of cancer.This is achieved by elucidating the dynamic interplay between systemic and cellular ATP homeostasis,deciphering the intricate mechanisms governing AICD,elucidating its intricate involvement in cancer signaling pathways,and scrutinizing validated key genes.Moreover,the exploration of AICD as a potential avenue for cancer treatment underscores its essential role in shaping the future landscape of cancer therapeutics.
基金supported by the National Natural Science Foundation of China,No.81371346,81271376
文摘After hypoxia, ischemia, or inflammatory injuries to the central nervous system, the damaged cells release a large amount of adenosine triphosphate, which may cause secondary neuronal death. Autophagy is a form of cell death that also has neuroprotective effects. Cell Counting Kit assay, monodansylcadaverine staining, flow cytometry, western blotting, and real-time PCR were used to determine the effects of exogenous adenosine triphosphate treatment at different concentrations (2, 4, 6, 8, 10 mmol/L) over time (1, 2, 3, and 6 hours) on the apoptosis and autophagy of SH-SY5Y cells. High concentrations of extracellular adenosine triphosphate induced autophagy and apoptosis of SH-SYSY cells. The enhanced autophagy first appeared, and peaked at 1 hour after treatment with adenosine triphosphate. Cell apoptosis peaked at 3 hours, and persisted through 6 hours. With prolonged exposure to the adenosine triphosphate treatment, the fraction of apoptotic cells increased. These data suggest that the SH-SY5Y neural cells initiated autophagy against apoptosis within an hour of adenosine triphosphate treatment to protect themselves against injury.
文摘The mammalian target of rapamycin (mTOR) pathway plays an important role in neuronal growth, proliferation and differentiation. To better understand the role of mTOR pathway involved in the induction of spinal cord injury, rat models of spinal cord injury were established by modified Allen's stall method and interfered for 7 days by intraperitoneal administration of mTOR activator adenosine triphosphate and mTOR kinase inhibitor rapamycin. At 1-4 weeks after spinal cord injury induction, the Basso, Beattie and Bresnahan locomotor rating scale was used to evaluate rat locomotor function, and immunohistochemical staining and western blot analysis were used to detect the expression of nestin (neural stem cell marker), neuronal nuclei (neuronal marker), neuron specific enolase, neurofilament protein 200 (axonal marker), glial fibrillary acidic protein (astrocyte marker), Akt, mTOR and signal transduction and activator of transcription 3 (STAT3). Results showed that adenosine triphosphate-mediated Akt/mTOR/STAT3 pathway increased endogenous neural stem cells, induced neurogenesis and axonal growth, inhibited excessive astrogliosis and improved the locomotor function of rats with spinal cord injury.
基金The National Natural Science Foundation of China(Grant No.21576134,Grant No.21606127,Grant No.21390200,Grant No.21706126)the National Key Research and Development Program of China(Grant No.2016YFA0204300)the Top-notch Academic Programs Project of Jiangsu Higher Education Institutions。
文摘In this study,we aimed at developing an efficient biocatalytic process for bio-production of cyclic adenosine monophosphate(c AMP)from adenosine triphosphate(ATP).First,adenylate cyclase from Escherichia coli MG1655(EAC)and Bordetella Pertussis(BAC)were expressed in E.coli BL21(DE3)and comparatively analyzed for their activities.As a result,EAC from E.coli MG1655 exhibited a higher activity.However,amount of EAC were obtained in an insoluble form.Therefore,we expressed the first 446 amino acids of EAC(EAC446)to avoid the inclusion body.The effects of induction temperature,incubation time,and incubation p H were further evaluated to improve the expression of EAC446.Subsequently,the reaction process for the production of c AMP with ATP as a starting material was investigated.As none of c AMP was detected in the whole-cell based biocatalytic process,the reaction catalyzed by the crude enzyme was determined for c AMP production.What's more,the reaction temperature,reaction p H,metal ion additives and substrate concentration was optimized,and the maximum c AMP production of 18.45 g·L^-1was achieved with a yield of 95.4%after bioconversion of 6 h.
文摘Nicotinamide adenine dinucleotide (NAD) oscillation was observed when the isolated mitochondria were immersed in a pyruvate solution. In addition, when an adenosine diphosphate (ADP) was added to the mitochondrial suspension containing pyruvate, adenosine triphosphate (ATP) oscillation was observed as well as NADH oscillation. At this time, the pH within mitochondria also oscillated. It was found that the oscillatory reaction of NADH caused by the membrane permeation of pyruvate continues, causing the oscillation of NADH and H+ in the subsequent reactions. The pH oscillation led to the ATP oscillation. It is considered that the oscillatory reaction caused by the gradual entry of pyruvate into mitochondria was thought to be carried over to both the citric acid cycle and the respiratory chain, ultimately leading to the ATP oscillation in oxidative phosphorylation. Similarly, it was found that membrane permeation of malate causes the gradual occurrence of NADH, at which point NADH oscillates, followed by an oscillatory reaction of the respiratory chain, and finally ATP oscillation. It was found that the oscillations of NADH and ATP occur without going through the citric acid cycle. Oscillations of NADH and other intermediates in both the citric acid cycle and respiratory chain were also confirmed by experiments using semipermeable membranes. These results support our hypothesis that the gradual entry of the substrate by membrane permeation triggers an oscillatory reaction of the enzyme, which is also carried over to subsequent reactions.
基金The study was supported by grants from the National Natural Science Foundation of China (No. 30470617 and No. 30570618).
文摘Background Repetitive transcranial magnetic stimulation (rTMS) research has mainly been focused on the therapeutic effect of psychiatric disorders and Parkinson's disease. A few studies have shown that rTMS might protect against delayed neuronal death induced by transient ischemia, enhance long-term potentiation in ischemic conditions and affect regional brain blood flow and metabolism. The aim of this study was to determine the effects of repetitive transcranial magnetic stimulation (rTMS) on adenosine triphosphate (ATP) content and microtubule associated protein-2 (MAP-2) expression in rat brain after middle cerebral artery occlusion (MCAO)/reperfusion. Methods To study the effects of different timecourses of rTMS on ATP content and MAP-2 expression, 90 rats were randomly divided into three groups (30 rats in each group). To study the effects of multiple rTMS parameters on ATP content and MAP-2 expression, the rats in each group were further divided into six subgroups (five rats each). The rats were sacrificed at 1-hour, 24-hour and 48-hour intervals after reperfusion, and the brain tissues were collected for the detection of ATP and MAP-2. Results rTMS could significantly increase ATP content and MAP-2 expression in the left brain following ischemic insult (P 〈0.01) and different rTMS parameters had different effects on the ATP level and the MAP-2 expression in the left striatum. A high-frequency rTMS played an important role in MAP-2 expression and ATP preservation. Conclusions This study revealed that rTMS induced significant increase of ATP content and MAP-2 expression in the injured area of the brain, suggesting that the regulation of both ATP and MAP-2 may be involved in the biological mechanism of the effect of rTMS on neural recovery. Therefore, rTMS may become a potential adjunctive therapy for ischemic cerebrovascular disease.
基金the National Natural Science Foundation of China(Nos.21701015,61975214,21811530054,61671435)Beijing Natural Science Foundation(No.4202075)the National Key R&D Program(No.2018YFC0115500)。
文摘In this work,polymethacrylic acid(PMAA)-templated silver nanoclusters(Ag NCs)were developed as the fluorescent probe for the efficient and sensitive detection of adenosine triphosphate(ATP)in a wide range of pH values.The fluorescence intensity of the Ag NCs could keep stable with pH values ranging from2.5 to 9.3.The detection of ATP was based on the quenching of the fluorescent Ag NCs in the presence of ATP.The fluorescence quenching of the Ag NCs with increasing ATP concentration was studied at pH 2.5,4.5,7.0 and 8.5 which involved a wide pH environment in body fluids.The limit of detection(LOD)for ATP was as low as 0.1 mmol/L in an acidic environment with pH of 2.5 and all the linear correlation coefficients were satisfactory under wide-span pH values from 2.5 to 8.5.In addition,the sensitive determination of ATP was also achieved by adding copper ions(Cu^2+).The high selectivity and rapid detection process proved that the fluorescent probe had great potential to detect ATP in biological samples under different pH conditions.
基金The authors gratefully acknowledge the financial support of the Natural Science Foundation of China(nos.21735002,21575037,21778016,21675046,and 21877030).
文摘Transmembrane transport plays an important role in many physiological functions,and mimicking this biological process in artificial systems has potential applications in biosensing,drug delivery,and bionic science.Here,a lipophilic split aptamer was developed as a novel transmembrane carrier for adenosine triphosphate(ATP)transport.The ATP carrier comprises two split aptamer fragments and cholesterol tags,with the split aptamers acting as targetrecognition domains to enhance their specific binding capability and the cholesterol tags as hydrophobic domains to facilitate membrane penetration.Giant unilamellar vesicle experiments demonstrated that the ATP carrier-mediated transmembrane transport was concentration-and time-dependent and showed high transport selectivity.Moreover,the artificial carriers were applicable to living cells and facilitated rapid cell internalization of fluorescencelabeled ATP.Furthermore,carrier-mediated ATP transport into ATP-deficient cells enabled recovery of cellular ATP levels and improved cell viability.This study demonstrated the efficacy of an aptamer nanostructure for designing DNA-based synthetic carriers with high selectivity and flexibility.
基金supported by the National Natural Science Foundation of China,No.81273926,81573742the Natural Science Foundation of Zhejiang Province of China,No.LY15H290006
文摘The activation of adenosine A1 receptors is important for protecting against ischemic brain injury and pretreatment with electroacupuncture has been shown to mitigate ischemic brain insult. The aim of this study was to test whether the adenosine A1 receptor mediates electroacupuncture pretreatment-induced neuroprotection against ischemic brain injury. We first performed 30 minutes of electroacupuncture pretreatment at the Baihui acupoint(GV20), delivered with a current of 1 mA, a frequency of 2/15 Hz, and a depth of 1 mm. High-performance liquid chromatography found that adenosine triphosphate and adenosine levels peaked in the cerebral cortex at 15 minutes and 120 minutes after electroacupuncture pretreatment, respectively. We further examined the effect of 15 or 120 minutes electroacupuncture treatment on ischemic brain injury in a rat middle cerebral artery-occlusion model. We found that at 24 hours reperfusion,120 minutes after electroacupuncture pretreatment, but not for 15 minutes, significantly reduced behavioral deficits and infarct volumes. Last, we demonstrated that the protective effect gained by 120 minutes after electroacupuncture treatment before ischemic injury was abolished by pretreatment with the A1-receptor antagonist 8-cyclopentyl-1,3-dipropylxanthine(1 mg/kg, intraperitoneally). Our results suggest that pretreatment with electroacupuncture at the Baihui acupoint elicits protection against transient cerebral ischemia via action at adenosine A1 receptors.
基金supported by a grant from PICT2019-N°01665 to HRQ
文摘Traumatic spinal cord injuries interrupt the connection of all axonal projections with their neuronal targets below and above the lesion site. This interruption results in either temporary or permanent alterations in the locomotor, sensory, and autonomic functions. Damage in the spinal tissue prevents the re-growth of severed axons across the lesion and their reconnection with neuronal targets. Therefore, the absence of spontaneous repair leads to sustained impairment in voluntary control of movement below the injury. For decades, axonal regeneration and reconnection have been considered the opitome of spinal cord injury repair with the goal being the repair of the damaged long motor and sensory tracts in a complex process that involves:(1) resealing injured axons;(2) reconstructing the cytoskeletal structure inside axons;(3) re-establishing healthy growth cones;and(4) assembling axonal cargos. These biological processes require an efficient production of adenosine triphosphate, which is affected by mitochondrial dysfunction after spinal cord injury. From a pathological standpoint, during the secondary stage of spinal cord injury, mitochondrial homeostasis is disrupted, mainly in the distal segments of severed axons. This result in a reduction of adenosine triphosphate levels and subsequent inactivation of adenosine triphosphate-dependent ion pumps required for the regulation of ion concentrations and reuptake of neurotransmitters, such as glutamate. The consequences are calcium overload, reactive oxygen species formation, and excitotoxicity. These events are intimately related to the activation of necrotic and apoptotic cell death programs, and further exacerbate the secondary stage of the injury, being a hallmark of spinal cord injury. This is why restoring mitochondrial function during the early stage of secondary injury could represent a potentially effective therapeutic intervention to overcome the motor and sensory failure produced by spinal cord injury. This review discusses the most recent evidence linking mitochondrial dysfunction with axonal regeneration failure in the context of spinal cord injury. It also covers the future of mitochondria-targeted therapeutical approaches, such as antioxidant molecules, removing mitochondrial anchor proteins, and increasing energetic metabolism through creatine treatment. These approaches are intended to enhance functional recovery by promoting axonal regenerationreconnection after spinal cord injury.
基金supported by grants of the Administrative Bureau of Chinese Traditional Medicine of Guangdong Province of China,No.2007109the Medical Research Foundation of Guangdong Province of China,No.A20111154
文摘In our previous study, we found that the edible alcohol extract of the root of the medicinal plant Rhodiola crenulata(RCE) improved spatial cognition in a rat model of Alzheimer's disease. Another study from our laboratory showed that RCE enhanced neural cell proliferation in the dentate gyrus of the hippocampus and prevented damage to hippocampal neurons in a rat model of chronic stress-induced depression. However, the mechanisms underlying the neuroprotective effects of RCE are unclear. In the present study, we investigated the anti-apoptotic effect of RCE and its neuroprotective mechanism of action in a rat model of Alzheimer's disease established by intracerebroventricular injection of streptozotocin. The rats were pre-administered RCE at doses of 1.5, 3.0 or 6.0 g/kg for 21 days before model establishment. ATP and cytochrome c oxidase levels were significantly decreased in rats with Alzheimer's disease. Furthermore, neuronal injury was obvious in the hippocampus, with the presence of a large number of apoptotic neurons. In comparison, in rats given RCE pretreatment, ATP and cytochrome c oxidase levels were markedly increased, the number of apoptotic neurons was reduced, and mitochondrial injury was mitigated. The 3.0 g/kg dose of RCE had the optimal effect. These findings suggest that pretreatment with RCE prevents mitochondrial dysfunction and protects hippocampal neurons from apoptosis in rats with Alzheimer's disease.
文摘Aim: To investigate the possible role of manganese in the regulation of mitochondrial aconitase (mACON) activity human prostate carcinoma cell line PC-3 cells. Methods: The mACON enzymatic activities of human prostate carcinoma cell line PC-3 cells were determined using a reduced nicotinamide adenine dinucleotide-coupled assay. Immunoblot and transient gene expression assays were used to study gene expression of the mACON. The putative response element for gene expression was identified using reporter assays with site-directed mutagenesis and electrophoretic mobility-shift assays. Results: In vitro study revealed that manganese chloride (MnCI2) treatment for 16 h inhibited the enzymatic activity of mACON, which induced the inhibition of citrate utility and cell proliferation of PC- 3 cells. Although results from transient gene expression assays showed that MnCI2 treatment upregulated gene translation by approximately 5-fold through the iron response element pathway, immunoblot and reporter assays showed that MnCl2 treatments inhibited protein and gene expression of mACON. This effect was reversed by cotreatment with ferric ammonium citrate. Additional reporter assays with site-directed mutagenesis and electrophoretic mobility-shift assays suggested that a putative metal response element in the promoter of the mACON gene was involved in the regulation of MnCh on the gene expression of mACON. Conclusion: These findings suggest that manganese acts as an antagonist of iron, disrupting the enzymatic activity and gene expression of mACON and citrate metabolism in the prostate.
基金supported by the National Natural Science Foundation of China (No. 81072505)
文摘Summary: Relationship between ATP changes of rabbit blood and postmortem interval (PMI) was studied. Twenty-four healthy rabbits were sacrificed and randomly divided into 3 groups with 8 rab- bits of each group. The bodies of three groups were placed in calorstat at temperature of 15℃, 25℃ and 35℃, respectively. The blood from the right ventricle was sampled through indwelling needle each 4 h until 72 h after death. ATP levels in the blood samples were measured by using ATP fluo- rescence rapid detection technique at different PMIs. Blood ATP levels slightly increased in the early stage after death and then constantly declined at all temperatures (15℃, 25℃, and 35℃). Cubic polynomial regression equations with log[ATP] as dependent variable (y) and PMI as independent variable (x) at different temperatures and the optimal time period were established as followed: Under 15℃ and during 16-64 h after death, y=-3.027×10^-5x^3+0.003x^2-0.096x-10.625 (Ra^2=0.992, P〈0.001); under 25℃ and during 8-56 h after death, y=-2.921×10^-5x^3+0.002x^2- 0.059x-11.186 (Ra^2=0.989, P〈0.001); under 35℃ and during 4-36 h after death, y=-9.769×10^-5x^3+ 0.005x^2 -0.117x-11.166 (Ra^2=0.991, P〈0.001). The changes in ATP levels in blood collected from right ven- tricle of rabbit cadavers showed relatively stable and regular degradation within 72 h after death at different temperatures.
基金Supported by National Natural Science Foundation of China(No.81100665)
文摘AIM: To investigate the potential of pigment epitheliumderived factor(PEDF) to protect the immortalized rat retinal ganglion cells-5(RGC-5) exposed to Co Cl2-induced chemical hypoxia. METHODS: After being differentiated with staurosporine(SS), RGC-5 cells were cultured in four conditions: control group cells cultured in Dulbecco 's modified eagle medium(DMEM) supplemented with 10% fetal bovine serum, 100 μmol/m L streptomycin and penicillin(named as normal conditions); hypoxia group cells cultured in DMEM containing 300 μmol/m L Co Cl2; cells in the group protected by PEDF were first pretreated with 100 ng/m L PEDF for 2h and then cultured in the same condition as hypoxia group cells; and PEDF group cells that were cultured in the presence of 100 ng/m L PEDF under normal conditions. The cell viability was assessed by MTT assay, the percentage of apoptotic cells was quantified using Annexin V-FITC apoptosis kit, and intra-cellar reactive oxygen species(ROS) was measured by dichloro-dihydro-fluorescein diacetate(DCFH-DA) probe. The mitochondria-mediated apoptosis was also examined to further study the underlying mechanism of the protective effect of PEDF. The opening of mitochondrial permeability transition pores(m PTPs) and membrane potential(Δψm) were tested as cellular adenosine triphosphate(ATP) level and glutathione(GSH). Also, the expression and distribution of Cyt C and apoptosis inducing factor(AIF) were observed.RESULTS: SS induced differentiation of RGC-5 cells resulting in elongation of their neurites and establishing contacts between outgrowths. Exposure to 300 μmol/m L Co Cl2 triggered death of 30% of the total cells in cultures within 24 h. At the same time, pretreatment with 100 ng/m L PEDF significantly suppressed the cell death induced by hypoxia(P〈0.05). The apoptosis induced by treatment of Co Cl2 was that induced cell death accompanied with increasing intracellar ROS and decreasing GSH and ATP level. PEDF pretreatment suppressed these effects(P〈0.05). Additionally, PEDF treatment inhibited the opening of m PTPs and suppressed decreasing of Δψm in RGC-5 cells, resulting in blocking of the mitochondrial apoptotic pathway.CONCLUSION: Pretreatment of RGC-5 cells with 100 ng/m L PEDF significantly decreases the extent of apoptosis. PEDF inhibits the opening of m PTPs and suppresses decreasing of Δψm. Moreover, PEDF also reduces ROS production and inhibits cellular ATP level's reduction. Cyt C and AIF activation in PEDF-pretreated cultures are also reduced. These results demonstrate the potential for PEDF to protect RGCs against hypoxic damage in vitro by preventing mitochondrial dysfunction.
文摘AIM:To measure the expression of transient receptor potential cation channel subfamily V member 4(TRPV4)in the rat cornea and determine whether it is related to adenosine triphosphate(ATP)generation in a rat model of acute ocular hypertension(AOH).METHODS:Immunofluorescence staining of TRPV4,P2X2 receptor,P2X3 receptor,and(33-tubulin in rat corneal longitudinal sections and paved was performed to clearly display histological structures.Rat models of AOH and agonist/antagonist-treated groups were established and corneal ATP was measured using an ATP assay.The independent t-test and simple linear correlation model were adopted for statistical analyses.RESULTS:Immunofluorescence staining of rat cornea sections revealed that epithelial and endothelial membranes showed strong immunoreactivity for TRPV4 and P2X2 receptor and coexpression with(33-tubulin in the rat corneal epithelial layer.Corneal ATP was significantly higher in the AOH rat model than in the control(P<0.05)and apparently lower after pretreatment by applying eyedrops of TRPV4 antagonist RN1734 with 30-40 mm Hg intraocular pressure(IOP;P<0.05).A simple linear regression model showed a positive correlation between rat corneal ATP and IOP values(R^(2)=0.996,P=0.0134)from the normal IOP(113 mm Hg)to 40 mm Hg.At 10-40min after anterior chamber injection of GSK1016790A(0.01 mL,50 nmol/L in 0.9%NaCl),corneal ATP was significantly higher than in the control group(P<0.05),which peaked at 1Omin.The ATP concentration of the normal epithelium was higher than that of the endothelium in the AOH rat model and after anterior chamber injection of GSK1016790A(P<0.05).CONCLUSION:The ATP concentration in the AOH rat cornea is increased by TRPV4 activation.
基金supported by grants from the Special Doctor Program of Nantong University(No.05024276)the Outstanding Teacher Program of Nantong University(No.03080542)
文摘This study investigated the protective effect of ATP on skeletal muscle satellite cells damaged by H2O2 in neonatal rats and the possible mechanism. The skeletal muscle satellite cells were randomly divided into four groups: normal group, model group(cells treated with 0.1 mmol/L H2O2 for 50 s), protection group(cells treated with 16, 8, 4, 2, 1, 0.5, or 0.25 mmol/L ATP for 24 h, and then with 0.1 mmol/L H2O2 for 50 s), proliferation group(cells treated with 16, 8, 4, 2, 1, 0.5, or 0.25 mmol/L ATP for 24 h). MTT assay, FITC+PI+DAPI fluorescent staining, Giemsa staining and immunofluorescence were performed to examine cell viability and apoptosis, and apoptosis-related proteins. The results showed that the survival rate of skeletal muscle satellite cells was decreased and the apoptosis rate was increased after H2O2 treatment(P〈0.01). Different doses of ATP had different effects on skeletal muscle satellite cells damaged by H2O2: the survival rate of muscle satellite cells treated with ATP at 4, 2, or 1 mmol/L was increased. The protective effect was most profound on cells treated with 2 mmol/L ATP. Immunofluorescence showed that ATP could increase the number of Bcl-2-positive cells(P〈0.01) and decrease the number of the Bax-positive cells(P〈0.01). It was concluded that ATP could protect skeletal muscle satellite cells against H2O2 damage in neonatal rats, which may be attributed to the up-regulation of the expression of Bcl-2 and down-regulation of Bax, resulting in the suppression of apoptosis.
基金the National Natural Science Foundation of China(No.10472027) the NUS Academic Research Fund(No.R-263-000-483-112)
文摘In consideration of the mechanism for shear-stress-induced Ca^2+ influx via ATP(adenosine triphosphate)-gated ion channel P2X4 in vascular endothelial cells, a modified model is proposed to describe the shear-stress-induced Ca^2+ influx. It is affected both by the Ca^2+ gradient across the cell membrane and extracellular ATP concentration on the cell surface. Meanwhile, a new static ATP release model is constructed by using published experimental data. Combining the modified intracellular calcium dynamics model with the new ATP release model, we establish a nonlinear Ca^2+ dynamic system in vascular endothelial cells. The ATP-mediated calcium response in vascular endothelial cells subjected to shear stresses is analyzed by solving the governing equations of the integrated dynamic system. Numerical results show that the shear-stress-induced calcium response predicted by the proposed model is more consistent with the experimental observations than that predicted by existing models.
文摘The paper introduces a new type instrument for rapid measuring ATP.The system consists of a micromodule ATP sensor and an instrument for measuring weak light transmitted by optic fiber. The micromodule ATP sensor mainly is composed of enzyme membrane, a probe and a bundle of optic fiber. The instrument measuring weak light consists of photomultiplier, high voltage power, pulse amplifier and counter. The instrument was characterized by simple structure,small size, rapid response time (<5 s), high sensitivity (10-12 mol/L), stable performance (measuring the same sample for 50 times, CV< 5% ), long enzyme storage time (>3 months).
基金Supported by a grant from the Korea Health Technology R&D Project through the Korea Health Industry Development Institute(KHIDI)(funded by the Ministry of Health&Welfare of the Republic of Korea),No.HI20C1405.
文摘BACKGROUND Adrenoleukodystrophy(ALD)is caused by a deficit in the ABCD1 gene,which leads to demyelination of neurons and dysfunction of the adrenal cortices and testicles.Of the three known phenotypes,30%-50%of male ALD patients present with the adrenomyeloneuropathy phenotype,characterized by gait disturbance as the initial symptom.CASE SUMMARY A 46-year-old man with a prior diagnosis of ALD was admitted to a Korean medicine hospital for the treatment of gait disturbance.His ability to walk was severely impaired at admission,significantly affecting the patient’s quality of life.He was treated with acupuncture,pharmacopuncture,electroacupuncture,and herbal medicine for 23 d.The 25-Foot Walk test(25FW),timed up and go(TUG),comfortable gait speed(CGS),numeric rating scale(NRS),Berg Balance Scale(BBS),Tinetti test,manual muscle test(MMT),and 3-level version of EuroQol-5 dimension(EQ-5D-3L)were used to evaluate the patient.The outcomes of the 25FW,TUG,and CGS improved during hospitalization.From the time of admission to discharge we observed:A decrease in NRS scores in the lower extremities and the lower back;an increase of 3 points in the BBS;a 1-point increase in the balancing part of the Tinetti Test;MMT and EQ-5D-3L performances remained unchanged.CONCLUSION Traditional Chinese medicine treatments could be a therapeutic option to alleviate issues related to gait disturbance in ALD.
基金Guangxi Natural Science(2018GXNSFBA-050051)Doctor Initiation Fund Project of Guangxi University of Traditional Chinese Medicine(2020BS011)。
文摘[Objectives]To detect the contents of components in Shuanghuanglian freeze-dried powder and to explore the effects of Shuanghuanglian freeze-dried powder and its components on human hepatocytes(HL-7702)alone or in combination with cefradine.[Methods]High performance liquid chromatography(HPLC)was used to detect the contents of baicalin,wogonin,chlorogenic acid and forsythin,the main components of Shuanghuanglian freeze-dried powder.HL-7702 cells were cultured with Shuanghuanglian freeze-dried powder and the main components of Shuanghuanglian freeze-dried powder alone or in combination with cefradine.Enzyme linked immunosorbent assay(ELISA)was used to detect the contents of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in the cell supernatant after culture,and HPLC was used to detect the expression level of adenosine diphosphate(ADP)and adenosine triphosphate(ATP);agarose gel electrophoresis was used to detect the expression of cyclooxygenase 2(COX-2)and heme oxygenase-1(HO-1)in HL-7702 cells.[Results]In Shuanghuanglian freeze-dried powder for injection,the content of baicalin was the highest,and the content of wogonin was the lowest.Compared with the control group,the expressions of AST and ALT in human hepatocytes(HL-7702)in high-dose baicalin group,forsythin group and wogonin group decreased(P<0.05,P<0.01),while the expression of ALT in chlorogenic acid+cefradine group(0.046 mg/mL)and forsythin+cefradine group(0.046 mg/mL)increased(P<0.05,P<0.01),and the expression of AST had no significant difference(P>0.05);the results in the low-dose group were similar to those in the high-dose group.Compared with the control group,ATP expression in chlorogenic acid group,chlorogenic acid+cefradine group(0.046 mg/mL)and forsythin+cefradine group(0.046 mg/mL)in the high-dose group decreased(P<0.05,P<0.01),and ADP expression was not significantly different(P>0.05);in the low-dose group,the expression of ATP and ADP increased in baicalin group(P<0.05),but decreased in wogonin group,baicalin+cefradine group(0.046 mg/mL)and wogonin group+cefradine group(0.046 mg/mL)(P<0.05,P<0.01).Compared with the control group,the expressions of COX-2 and HO-1 in HL-7702 cells in the cefradine group showed no significant difference(P>0.05).The expression of HO-1 and COX-2 in the different dose groups of Shuanghuanglian and the group combined with cefradine increased,and the difference was significant(P<0.05,P<0.01).[Conclusions]The components of Shuanghuanglian freeze-dried powder for injection had effects on hepatocytes,of which baicalin had a significant effect,and the effect of cefradine on hepatocytes was increased when used in combination with cefradine.