Inflammatory markers,oxidative stress,and mitochondrial dynamics:Repercussions on coronary artery disease in diabetes

Inflammatory markers and mediators that affect the development of cardiovascular diseases have been the focus of recent scientific work.Thus,the purpose of this editorial is to promote a critical debate about the article titled“Nε-carboxymethyl-lysine and inflammatory cytokines,markers,and mediators of coronary artery disease progression in diabetes”,published in the World Journal of Diabetes in 2024.This work directs us to reflect on the role of advanced glycation end products,which are pro-inflammatory products arising from the metabolism of fatty acids and sugars whose main marker in tissues is Nε-carboxymethyllysine(NML).Recent studies have linked high levels of pro-inflammatory agents with the development of coronary artery disease(CAD),especially tumor necrosis factor alpha,interleukins,and C-reactive protein.These inflammatory agents increase the production of reactive oxygen species(ROS),of which people with diabetes are known to have an increased production.The increase in ROS promotes lipid peroxidation,which causes damage to myocytes,promoting myocardial damage.Furthermore,oxidative stress induces the binding of NML to its receptor RAGE,which in turn activates the nuclear factor-kB,and consequently,inflammatory cytokines.These inflammatory cytokines induce endothelial dysfunction,with increased expression of adhesion molecules,changes in endothelial permeability and changes in the expression of nitric oxide.In this sense,the therapeutic use of monoclonal antibodies(inflammatory reducers such as statins and sodium-glucose transport inhibitors)has demonstrated positive results in the regression of atherogenic plaques and consequently CAD.On the other hand,many studies have demonstrated a relationship between mitochondrial dynamics,diabetes,and cardiovascular diseases.This link occurs since ROS have their origin in the imbalance in glucose metabolism that occurs in the mitochondrial matrix,and this imbalance can have its origin in inadequate diet as well as some pathologies.Photobiomodulation(PBM)has recently been considered a possible therapeutic agent for cardiovascular diseases due to its effects on mitochondrial dynamics and oxidative stress.In this sense,therapies such as PBM that act on pro-inflammatory mediators and mitochondrial modulation could benefit those with cardiovascular diseases.

Changes of mitochondrial dynamics during the progression of non-alcoholic fatty liver disease: a review

Non-alcoholic fatty liver disease(NAFLD)is a chronic liver disease closely related to metabolic disorders that pose a serious threat to human health.Currently,no specific drugs are available for treating the aetiology of NAFLD in clinical practice.Mitochondria have various biological functions inside the cell.Studies have found that mitochondrial fission and fusion are closely related to NAFLD.Therefore,identifying therapeutic targets for NAFLD through mitochondrial fission and fusion is crucial.Particularly in the field of traditional Chinese medicine,good therapeutic effects have been achieved in the treatment of NAFLD by protecting mitochondrial fusion and fission.Therefore,this article reviews the relationship between mitochondrial dynamics and NAFLD as well as the treatment of NAFLD through the regulation of mitochondrial fission and fusion with traditional Chinese medicine to provide a reference for the clinical application of traditional Chinese medicine in regulating mitochondrial fission and fusion functions to treat NAFLD.

Hypoxia promotes pulmonary vascular remodeling via HIF-1α to regulate mitochondrial dynamics

Background Increasing research suggests that mitochondrial defect plays a major role in pulmonary hypertension(PH) pathogenesis. Mitochondrial dynamics and quality control have a central role in the maintenance of the cell proliferation and apoptosis balance. However, the molecular mechanism underlying of this balance is still unknown. Methods To clarify the biological effects of hypoxic air exposure and hypoxia-inducible factor-1α(HIF-1α) on pulmonary arterial smooth muscle cell(PASMC) and pulmonary arterial hypertension rats, the cells were cultured in a hypoxic chamber under oxygen concentrations. Cell viability, reactive oxygen species level, cell death, mitochondrial morphology, mitochondrial membrane potential, mitochondrial function and mitochondrial biosynthesis, as well as fission-and fusion-related proteins, were measured under hypoxic conditions. In addition, rats were maintained under hypoxic conditions, and the right ventricular systolic pressure, right ventricular hypertrophy index and right ventricular weight/body weight ratio were examined and recorded. Further, we assessed the role of HIF-1α in the development and progression of PH using HIF-1α gene knockdown using small interfering RNA transfection. Mdivi-1 treatment was performed before hypoxia to inhibit dynamin-related protein 1(Drp1). Results We found that HIF-1α expression was increased during hypoxia, which was crucial for hypoxia-induced mitochondrial dysfunction and hypoxia-stimulated PASMCs proliferation and apoptosis. We also found that targeting mitochondrial fission Drp1 by mitochondrial division inhibitor Mdivi-1 was effective in PH model rats. The results showed that mitochondrial dynamics were involved in the pulmonary vascular remodeling under hypoxia in vivo and in vitro. Furthermore, HIF-1α also modulated mitochondrial dynamics in pulmonary vascular remodeling under hypoxia through directly regulating the expression of Drp1. Conclusions In conclusion, our data suggests that abnormal mitochondrial dynamics could be a marker for the early diagnosis of PH and monitoring disease progression. Further research is needed to study the signaling pathways that govern mitochondrial fission/fusion in PH.

Human mesenchymal stem cells exhibit altered mitochondrial dynamics and poor survival in high glucose microenvironment

BACKGROUND Mesenchymal stem cells(MSCs)are a type of stem cells that possess relevant regenerative abilities and can be used to treat many chronic diseases.Diabetes mellitus(DM)is a frequently diagnosed chronic disease characterized by hyperglycemia which initiates many multisystem complications in the long-run.DM patients can benefit from MSCs transplantation to curb down the pathological consequences associated with hyperglycemia persistence and restore the function of damaged tissues.MSCs therapeutic outcomes are found to last for short period of time and ultimately these regenerative cells are eradicated and died in DM disease model.AIM To investigate the impact of high glucose or hyperglycemia on the cellular and molecular characteristics of MSCs.METHODS Human adipose tissue-derived MSCs(hAD-MSCs)were seeded in low(5.6 mmol/L of glucose)and high glucose(25 mmol/L of glucose)for 7 d.Cytotoxicity,viability,mitochondrial dynamics,and apoptosis were deplored using specific kits.Western blotting was performed to measure the protein expression of phosphatidylinositol 3-kinase(PI3K),TSC1,and mammalian target of rapamycin(mTOR)in these cells.RESULTS hAD-MSCs cultured in high glucose for 7 d demonstrated marked decrease in their viability,as shown by a significant increase in lactate dehydrogenase(P<0.01)and a significant decrease in Trypan blue(P<0.05)in these cells compared to low glucose control.Mitochondrial membrane potential,indicated by tetramethylrhodamine ethyl ester(TMRE)fluorescence intensity,and nicotinamide adenine dinucleotide(NAD+)/NADH ratio were significantly dropped(P<0.05 for TMRE and P<0.01 for NAD+/NADH)in high glucose exposed hAD-MSCs,indicating disturbed mitochondrial function.PI3K protein expression significantly decreased in high glucose culture MSCs(P<0.05 compared to low glucose)and it was coupled with significant upregulation in TSC1(P<0.05)and downregulation in mTOR protein expression(P<0.05).Mitochondrial complexes I,IV,and V were downregulated profoundly in high glucose(P<0.05 compared to low glucose).Apoptosis was induced as a result of mitochondrial impairment and explained the poor survival of MSCs in high glucose.CONCLUSION High glucose impaired the mitochondrial dynamics and regulatory proteins in hAD-MSCs ensuing their poor survival and high apoptosis rate in hyperglycemic microenvironment.

Mitochondrial dynamics caused by QoIs and SDHIs fungicides depended on FgDnm1 in Fusarium graminearum

Fusarium head blight(FHB) caused by Fusarium graminearum is a devastating fungal disease on small grain cereal crops,because it reduces yield and quality and causes the mycotoxin contamination to the grain.Dynamins and dynamin-related proteins(DRPs) are large GTPase superfamily members,which are typically involved in the budding and division of vesicles in eukaryotic cells,but their roles in Fusarium spp.remain unexplored.Here,we found that FgDnm1,a DRP and homolog to Dnm1 in Saccharomyces cerevisiae,contributes to the normal fungal growth,sexual reproduction and sensitivity to fungicides.In addition,we found FgDnm1 co-localizes with mitochondria and is involved in toxisome formation and deoxynivalenol(DON) production.Several quinone outside inhibitors(QoIs) and succinate dehydrogenase inhibitors(SDHIs) cause fragmentated morphology of mitochondria.Importantly,the deletion of FgDnm1displays filamentous mitochondria and blocks the mitochondrial fragmentation induced by QoIs and SDHIs.Taken together,our studies uncover the effect of mitochondrial dynamics in fungal normal growth and how such events link to fungicides sensitivity and toxisome formation.Thus,we concluded that altered mitochondrial morphology induced by QoIs and SDHIs depends on FgDnm1.

Regulation of neural stem cell fate decisions by mitochondrial dynamics

Stem cells possess the ability to divide symmetrically or asymmet- rically to allow for maintenance of the stem cell pool or become committed progenitors and differentiate into various cell lineages. The unique self-renewal capabilities and pluripotency of stem cells are integral to tissue regeneration and repair （Oh et al., 2014）. Mul- tiple mechanisms including intracellular programs and extrinsic cues are reported to regulate neural stem cell （NSC） fate （Bond et al., 2015）. A recent study, published in Cell Stern Cell, identified a novel mechanism whereby mitochondrial dynamics drive NSC fate （Khacho et al., 2016）.

Electroacupuncture activates AMPKα1 to improve learning and memory in the APP/PS1 mouse model of early Alzheimer’s disease by regulating hippocampal mitochondrial dynamics

Objective Studies have shown that electroacupuncture(EA)can alleviate cognitive impairments from Alzheimer’s disease(AD)by regulating the expression of adenosine monophosphate-activated protein kinase(AMPK),but the specific mechanism involved remains to be elucidated.Therefore,this study explores the potential mechanism by which EA improves cognitive function from the perspective of mitochondrial dynamics.Methods The four-month-old transgenic mice with amyloid precursor protein(APP)/presenilin 1(PS1)and AMPKα1-subunit conditional knockout(AMPKα1-cKO)were used for experiments.To evaluate the effects of EA treatment on cognitive function,the T-maze and Morris water maze were used.In addition,chemical exchange saturation transfer,thioflavin staining,transmission electron microscopy,mitochondrial membrane potential,and Western blotting were used to examine the potential mechanisms underlying the effects of EA on APP/PS1 mice.Results Both APP/PS1 mice and AMPKα1-cKO mice exhibited dysfunction in mitochondrial dynamics accompanied by learning and memory impairment.Inactivation of the AMPK/peroxisome proliferator-activated receptor-γcoactivator-1α(PGC-1α)pathway increased pathological amyloid-β(Aβ)deposition and aggravated the dysfunction in mitochondrial dynamics.In addition,EA rescued learning and memory deficits in APP/PS1 mice by activating the AMPK/PGC-1αpathway,specifically by reducing pathological Aβdeposition,normalizing energy metabolism,protecting the structure and function of mitochondria,increasing the levels of mitochondrial fusion proteins,and downregulating the expression of fission proteins.However,the therapeutic effect of EA on cognition in APP/PS1 mice was hindered by AMPKα1 knockout.Conclusion The regulation of hippocampal mitochondrial dynamics and reduction in Aβdeposition via the AMPK/PGC-1αpathway are critical for the ability of EA to ameliorate cognitive impairment in APP/PS1 mice.

Characterization,expression dynamics,and potential function of OPA1 for regulation of mitochondrial morphology during spermiogenesis in Phascolosoma esculenta

Mitochondria undergo morphological changes during spermatogenesis in some animals.The mechanism and role of mitochondrial morphology regulation,however,remain somewhat unclear.In this study,we analyzed the molecular characteristics,expression dynamics and subcellular localization of optic atrophy protein 1(OPA1),a mitochondrial fusion and cristae maintenance-related protein,to reveal the possible regulatory mechanisms underlying mitochondrial morphology in Phascolosoma esculenta spermiogenesis.The full-length cDNA of the P.esculenta opa1 gene(Pe-opa1)is 3743 bp in length and encodes 975 amino acids.The Pe-OPA1 protein is highly conservative and includes a transmembrane domain,a GTPase domain,two helical bundle domains,and a lipid-interacting stalk.Gene and protein expression was higher in the coelomic fluid(a site of spermatid development)of male P.esculenta and increased first and then decreased from March to December.Moreover,their expression during the breeding stage was significantly higher than during the non-breeding stage,suggesting that Pe-OPA1 is involved in P.esculenta reproduction.The Pe-OPA1 protein was more abundant in components consisting of many spermatids than in components without,indicating that Pe-OPA1 mainly plays a role in the spermatid in coelomic fluid.Moreover,Pe-OPA1 was mainly detected in the spermatid mitochondria.Immunofluorescence experiments showed that the Pe-OPA1 are constitutively expressed and co-localized with mitochondria during spermiogenesis,suggesting its involvement in P.esculenta spermiogenesis.These results provide evidence for Pe-OPA1's involvement in the regulation of mitochondrial morphology during spermiogenesis.

The essential functions of mitochondrial dynamics in immune cells

Mitochondria are highly mobile organelles due to fission,fusion,transport,and mitophagy,and these processes are known as mitochondrial dynamics.Mitochondrial dynamics play an important role in energy production,cell division,cell differentiation,and cell death.In the past decade,numerous studies have revealed the importance of mitochondrial metabolism in immunity,and mitochondrial dynamics are essential for immune responses mediated by various cell types.In this review,we mainly discuss the role of mitochondrial dynamics in activation,differentiation,cytokine production,and the activity of related pathways in immune cells,particularly T cells,B cells,and other cells involved in the innate immune response.

Shenmai injection enhances cisplatin-induced apoptosis through regulation of Mfn2-dependent mitochondrial dynamics in lung adenocarcinoma A549/DDP cells

Aim:Chemoresistance is the biggest obstacle in cancer treatment.Our previous study demonstrated that Shenmai injection(SMI),a Chinese herbal medicine,enhanced the antitumor effect of cisplatin via glucose metabolism reprogramming.This study aimed to further determine whether the SMI sensitizes the non-small cell lung cancer(NSCLC)cells to cisplatin through regulation mitochondrial dynamics.Methods:The Kaplan-Meier Plotter database was used to investigate the relationship between mRNA expression of mitofusin-2(Mfn2)and the survival analysis of NSCLC patients.The protein expression of Mfn2 in a lung adenocarcinoma tissue chip was detected by immunohistochemistry staining.The expression of Mfn2 and ATAD3A were compared between cisplatin-sensitive A549 and cisplatin-resistant A549/DDP cells.Additionally,A549/DDP cells were co-treated with cisplatin and SMI to detect mitochondrial morphology by fluorescent staining,apoptosis-related protein expression with Western blotting,and mitochondrial membrane potential(ΔΨm)with flow cytometry analysis.Results:The mean survival time of the Mfn2^(low) group was significantly lower than that of the Mfn2^(high) group by Kaplan-Meier Plotter database analysis,and the Mfn2 protein expression level was lower in cancer tissues than in adjacent tissues.The combination of SMI and cisplatin induced dynamic changes in A549/DDP cells,with increased mitochondrial fusion followed by upregulation of Mfn2 and downregulation of ATAD3A and reduced mitochondrial mass and ΔΨm.Moreover,SMI significantly enhanced cisplatin-induced A549/DDP apoptosis,upregulated Bax and the active subunit of caspase-3,and downregulated Bcl-2 expression,as shown via Hoechst staining and flow cytometry analysis.Conclusion:Our findings suggest SMI enhances cisplatin-induced apoptosis through regulation of Mfn2-dependent mitochondrial dynamics in cisplatin-resistant lung adenocarcinoma cells.

Mitochondrial dysfunction and quality control lie at the heart of subarachnoid hemorrhage

The dramatic increase in intracranial pressure after subarachnoid hemorrhage leads to a decrease in cerebral perfusion pressure and a reduction in cerebral blood flow.Mitochondria are directly affected by direct factors such as ischemia,hypoxia,excitotoxicity,and toxicity of free hemoglobin and its degradation products,which trigger mitochondrial dysfunction.Dysfunctional mitochondria release large amounts of reactive oxygen species,inflammatory mediators,and apoptotic proteins that activate apoptotic pathways,further damaging cells.In response to this array of damage,cells have adopted multiple mitochondrial quality control mechanisms through evolution,including mitochondrial protein quality control,mitochondrial dynamics,mitophagy,mitochondrial biogenesis,and intercellular mitochondrial transfer,to maintain mitochondrial homeostasis under pathological conditions.Specific interventions targeting mitochondrial quality control mechanisms have emerged as promising therapeutic strategies for subarachnoid hemorrhage.This review provides an overview of recent research advances in mitochondrial pathophysiological processes after subarachnoid hemorrhage,particularly mitochondrial quality control mechanisms.It also presents potential therapeutic strategies to target mitochondrial quality control in subarachnoid hemorrhage.

Latest assessment methods for mitochondrial homeostasis in cognitive diseases

Mitochondria play an essential role in neural function,such as supporting normal energy metabolism,regulating reactive oxygen species,buffering physiological calcium loads,and maintaining the balance of morphology,subcellular distribution,and overall health through mitochondrial dynamics.Given the recent technological advances in the assessment of mitochondrial structure and functions,mitochondrial dysfunction has been regarded as the early and key pathophysiological mechanism of cognitive disorders such as Alzheimer’s disease,Parkinson’s disease,Huntington’s disease,mild cognitive impairment,and postoperative cognitive dysfunction.This review will focus on the recent advances in mitochondrial medicine and research methodology in the field of cognitive sciences,from the perspectives of energy metabolism,oxidative stress,calcium homeostasis,and mitochondrial dynamics(including fission-fusion,transport,and mitophagy).

Dual Oxidase Mutant Retards Mauthner-Cell Axon Regeneration at an Early Stage via Modulating Mitochondrial Dynamics in Zebrafish

Dual oxidase(duox)-deriyed reactive oxygen species(ROS)have been correlated with neuronal polarity,cerebellar development,and neuroplasticity.However,there have not been many comprehensive studies of the effect of individual duox isoforms on central-axon regenerationin vivo.Here,we explored this question in zebrafish,an excellent model organism for central-axon regeneration studies.In our research,mutation of the duox gene with CRISPR/Cas9 significantly retarded the singleaxon regeneration of the zebrafish Mauthner cell in vivo.Using deep transcriptome sequencing,we found that the expression levels of related functional enzymes in mitochondria were down-regulated in duox mutant fish.In vivo imaging showed that duox mutants had significantly disrupted mitochondrial transport and redox state in single Mauthner-cell axon.Our research data provide insights into how duox is involved in central-axon regeneration by changing mitochondrial transport.

Mitochondrial dysfunction in glaucomatous degeneration

Glaucoma is a kind of optic neuropathy mainly manifested in the permanent death of retinal ganglion cells(RGCs),atrophy of the optic nerve,and loss of visual ability.The main risk factors for glaucoma consist of the pathological elevation of intraocular pressure(IOP)and aging.Although the mechanism of glaucoma remains an open question,a theory related to mitochondrial dysfunction has been emerging in the last decade.Reactive oxygen species(ROS)from the mitochondrial respiratory chain are abnormally produced as a result of mitochondrial dysfunction.Oxidative stress takes place when the cellular antioxidant system fails to remove excessive ROS promptly.Meanwhile,more and more studies show that there are other common features of mitochondrial dysfunction in glaucoma,including damage of mitochondrial DNA(mt DNA),defective mitochondrial quality control,ATP reduction,and other cellular changes,which are worth summarizing and further exploring.The purpose of this review is to explore mitochondrial dysfunction in the mechanism of glaucomatous optic neuropathy.Based on the mechanism,the existing therapeutic options are summarized,including medications,gene therapy,and red-light therapy,which are promising to provide feasible neuroprotective ideas for the treatment of glaucoma.

The “mitochondrial stress responses”: the “Dr. Jekyll and Mr. Hyde” of neuronal disorders

Neuronal disorders are associated with a profound loss of mitochondrial functions caused by various stress conditions,such as oxidative and metabolic stress,protein folding or import defects,and mitochondrial DNA alteration.Cells engage in different coordinated responses to safeguard mitochondrial homeostasis.In this review,we will explore the contribution of mitochondrial stress responses that are activated by the organelle to perceive these dangerous conditions,keep them under control and rescue the physiological condition of nervous cells.In the sections to come,particular attention will be dedicated to analyzing how compensatory mitochondrial hyperfusion,mitophagy,mitochondrial unfolding protein response,and apoptosis impact human neuronal diseases.Finally,we will discuss the relevance of the new concept:the“mito-inflammation”,a mitochondria-mediated inflammatory response that is recently found to cover a relevant role in the pathogenesis of diverse inflammatory-related diseases,including neuronal disorders.

Aldehyde dehydrogenase 2 preserves mitochondrial morphology and attenuates hypoxia/reoxygenationinduced cardiomyocyte injury

BACKGROUND:Disturbance of mitochondrial fi ssion and fusion(termed mitochondrial dynamics)is one of the leading causes of ischemia/reperfusion(I/R)-induced myocardial injury.Previous studies showed that mitochondrial aldehyde dehydrogenase 2(ALDH2)conferred cardioprotective effect against myocardial I/R injury and suppressed I/R-induced excessive mitophagy in cardiomyocytes.However,whether ALDH2 participates in the regulation of mitochondrial dynamics during myocardial I/R injury remains unknown.METHODS:In the present study,we investigated the effect of ALDH2 on mitochondrial dynamics and the underlying mechanisms using the H9c2 cells exposed to hypoxia/reoxygenation(H/R)as an in vitro model of myocardial I/R injury.RESULTS:Cardiomyocyte apoptosis was significantly increased after oxygen-glucose deprivation and reoxygenation(OGD/R),and ALDH2 activation largely decreased the cardiomyocyte apoptosis.Additionally,we found that both ALDH2 activation and overexpression significantly inhibited the increased mitochondrial fission after OGD/R.Furthermore,we found that ALDH2 dominantly suppressed dynamin-related protein 1(Drp1)phosphorylation(Ser616)and adenosine monophosphate-activated protein kinase(AMPK)phosphorylation(Thr172)but not interfered with the expression levels of mitochondrial shaping proteins.CONCLUSIONS:We demonstrate the protective effect of ALDH2 against cardiomyocyte H/R injury with a novel mechanism on mitochondrial fission/fusion.

Photobiomodulation provides neuroprotection through regulating mitochondrial fission imbalance in the subacute phase of spinal cord injury

Increasing evidence indicates that mitochonarial lission imbalance plays an important role in derayed neuronal cell death. Our previous study round that photo biomodulation improved the motor function of rats with spinal cord injury.However,the precise mechanism remains unclear.To investigate the effect of photo biomodulation on mitochondrial fission imbalance after spinal cord injury,in this study,we treated rat models of spinal co rd injury with 60-minute photo biomodulation(810 nm,150 mW) every day for 14 consecutive days.Transmission electron microscopy results confirmed the swollen and fragmented alte rations of mitochondrial morphology in neurons in acute(1 day) and subacute(7 and 14 days) phases.Photo biomodulation alleviated mitochondrial fission imbalance in spinal cord tissue in the subacute phase,reduced neuronal cell death,and improved rat posterior limb motor function in a time-dependent manner.These findings suggest that photobiomodulation targets neuronal mitochondria,alleviates mitochondrial fission imbalance-induced neuronal apoptosis,and thereby promotes the motor function recovery of rats with spinal cord injury.

Restoration of Mitochondrial Structure and Function within Helicobacter pylori VacA Intoxicated Cells

The Helicobacter pylori vacuolating cytotoxin (VacA) is an intracellular, mitochondrial-targeting exotoxin that rapidly causes mitochondrial dysfunction and fragmentation. Although VacA targeting of mitochondria has been reported to alter overall cellular metabolism, there is little known about the consequences of extended exposure to the toxin. Here, we describe studies to address this gap in knowledge, which have revealed that mitochondrial dysfunction and fragmentation are followed by a time-dependent recovery of mitochondrial structure, mitochondrial transmembrane potential, and cellular ATP levels. Cells exposed to VacA also initially demonstrated a reduction in oxidative phosphorylation, as well as increase in compensatory aerobic glycolysis. These metabolic alterations were reversed in cells with limited toxin exposure, congruent with the recovery of mitochondrial transmembrane potential and the absence of cytochrome c release from the mitochondria. Taken together, these results are consistent with a model that mitochondrial structure and function are restored in VacA-intoxicated cells.

Visualizing the distribution of mitochondrial molecular mRNAs of cerebellar Purkinje cells by RNAscope

Objective To investigate the distribution of mitochondria-associated molecule mRNA in cerebellar Purkinje cells.Methods Transgenic technology was used to prepare Pcp2-tdTomato mouse labeled Purkinje cells.The mRNAs of mitochondria-associated molecules Mfn2,Mfn1,Ucp4,Drp1,Ucp2,Mcu and Nclx were detected in situ by RNAscope technique.Results The results of this study showed that Mfn2,Mfn1,Ucp4 and Drp1 were highly expressed in Purkinje cell body(Bin 3>10%).However,Ucp2,Mcu and Nclx were less expressed.In addition,Mfn2,Mfn1,Ucp4,Drp1,Ucp2,Mcu and Nclx were less expressed in dendrites.Conclusion Current research focuses on developing new and more specific molecules to regulate the activity of Purkinje cells and opening therapeutic windows for Purkinje cells related diseases.The molecule identification of potential drug targets,mechanisms of action,and structural basis of their activity will crucially promote preclinical development.

Electroacupuncture preconditioning protects against focal cerebral ischemia/reperfusion injury via suppression of dynamin-related protein 1

Electroacupuncture preconditioning at acupoint Baihui （GV20） can reduce focal cerebral ischemia/reperfusion injury. However, the precise protective mechanism remains unknown. Mitochondrial fission mediated by dynamin-related protein 1 （Drp1） can trigger neuronal apoptosis following cerebral ischemia/reperfusion injury. Herein, we examined the hypothesis that electroacupuncture pretreatment can regulate Drp1, and thus inhibit mitochondrial fission to provide cerebral protection. Rat models of focal cerebral ischemia/reperfusion injury were established by middle cerebral artery occlusion at 24 hours after 5 consecutive days of preconditioning with electroacupuncture at GV20 （depth 2 mm, intensity 1 mA, frequency 2/15 Hz, for 30 minutes, once a day）. Neurological function was assessed using the Longa neurological deficit score. Pathological changes in the ischemic penumbra on the injury side were assessed by hematoxylin-eosin staining. Cellular apoptosis in the ischemic penumbra on the injury side was assessed by terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling staining. Mitochondrial ultrastructure in the ischemic penumbra on the injury side was assessed by transmission electron microscopy. Drp1 and cytochrome c expression in the ischemic penumbra on the injury side were assessed by western blot assay. Results showed that electroacupuncture preconditioning decreased expression of total and mitochondrial Drp1, decreased expression of total and cytosolic cytochrome c, maintained mitochondrial morphology and reduced the proportion of apoptotic cells in the ischemic penumbra on the injury side, with associated improvements in neurological function. These data suggest that electroacupuncture preconditioning-induced neuronal protection involves inhibition of the expression and translocation of Drp1.