Role of cyclic guanosine monophosphate-adenosine monophosphate synthase-stimulator of interferon genes pathway in diabetes and its complications

Diabetes mellitus(DM)is one of the major causes of mortality worldwide,with inflammation being an important factor in its onset and development.This review summarizes the specific mechanisms of the cyclic guanosine monophosphate-adenosine monophosphate synthase(cGAS)-stimulator of interferon genes(STING)pathway in mediating inflammatory responses.Furthermore,it compre-hensively presents related research progress and the subsequent involvement of this pathway in the pathogenesis of early-stage DM,diabetic gastroenteropathy,diabetic cardiomyopathy,non-alcoholic fatty liver disease,and other complic-ations.Additionally,the role of cGAS-STING in autonomic dysfunction and intes-tinal dysregulation,which can lead to digestive complications,has been discuss-ed.Altogether,this study provides a comprehensive analysis of the research advances regarding the cGAS-STING pathway-targeted therapeutic agents and the prospects for their application in the precision treatment of DM.

Highly-sensitive electrochemiluminescence biosensor for detection of inosine monophosphate in meat based on graphdiyne/AuNPs/luminol nanocomposites

Inosine monophosphate(IMP),as a critical umami substance,is one of the most important indicators for evaluating the quality of meat products.Here,a sensitive electrochemiluminescence(ECL)biosensor based on graphdiyne(GDY)/AuNPs/luminol nanocomposites was constructed to detect IMP.The GDY/AuNPs/luminol nanocomposites were synthesized by using simple one-pot method.GDY utilized its 2D framework to disperse and fix gold nanoparticles,which inhibited the agglomeration of gold nanoparticles and greatly improved its stability and catalytic properties.Importantly,GDY/AuNPs/luminol nanocomposites showed excellent catalytic ability and superior ECL activity towards luminol-H_(2)O_(2) systems due to the synergistic effect of GDY and AuNPs.Under optimal conditions,the prepared biosensor exhibited a wide linear range from 0.01 g/L to 20 g/L,a satisfactory limit detection of 0.0013 g/L,as well as an excellent specificity.Moreover,we carried out the precise analysis of IMP in actual meat samples with acceptable results compared to the liquid chromatography.We believe that this work could offer an efficient ECL platform for accurate and reliable report of IMP levels,which is significant for maintaining food quality and safety.

Characteristics of Deposition of Inosine Monophosphate(IMP) and Intramuscular Fat(IMF) in Muscles of Jinghai Yellow Chicken and Its Crossbreeds

[Objective] This study aimed to investigate the characteristics of deposition of inosine monophosphate （IMP） and intramuscular fat （IMF） in muscles of Jinghai yellow chickens and its crossbred.[Method] The characteristics of IMP and IMF deposition of 112-day-old Jinghai yellow chickens （J×J） and its two different 70-day- old crossbreeds （J×B and B×B） were analyzed. The IMP content in breast muscle and leg muscle were determined by HPLC. [Result] The contents of IMP and cor- rected inosine monophosphate （IMPc） in breast muscle were significantly or ex- tremely significantly higher than that in leg muscle of the chickens in the three groups whether in male or female chickens （P〈0.05 or P〈0.01）. There were no sig- nificant difference in the contents of IMP and IMPc between hens and roosters （P〉 0.05）. The fresh degree of breast muscle and leg muscle was 96,11%-98.16% and 87.22%-93.07%, respectively. And the fresh degree of breast muscle was higher than that of leg muscle. In the three groups, the IMF content in leg muscle was significantly higher than that in breast muscle whether in male or female chickens （P〈0.05）. The contents of IMF in breast muscle and leg muscle were 0.36%-0.75% and 1.84%-2.38%, respectively. The iMP content in breast muscle of chickens in Bx J group was extremely significantly higher than that in breast muscle of chickens in JxJ group （P〈0.01）, but the contents of IMPc and iMF of breast muscle and leg muscle of the chickens in the three groups had no significant difference （P〉0.05）. [Conclusion] To sum up, the freshness and flavor significantly differ between the breast muscle and leg muscle of chickens, but show no significant difference among the three groups.

Effects of plant extract neferine on cyclic adenosine monophosphate and cyclic guanosine monophosphate levels in rabbit corpus cavernosum in vitro

Aim： To further investigate the relaxation mechanism of neferine （NED, a bis-benzylisoquinoline alkaloid extracted （isolated） from the green seed embryo of Nelumbo nucifera Gaertn in China, on rabbit corpus cavernosum tissue in vitro. Methods： The effects of Nef on the concentrations of cyclic adenosine monophosphate （cAMP） and cyclic guanosine monophosphate （cGMP） in isolated and incubated rabbit corpus cavernosum tissue were recorded using ^125I radioimmunoassay. Results： The basal concentration of cAMP in corpus cavernosum tissue was 5.67 ± 0.97 pmol/mg. Nef increased the cAMP concentration in a dose-dependent manner （P 〈 0.05）, but this effect was not inhibited by an adenylate cyclase inhibitor （cis-N-[2-phenylcyclopentyl]azacyclotridec-1-en-2-amine, MDL-12, 330A） （P 〉 0.05）. The accumulation of cAMP induced by prostaglandin Et （PGEt, a stimulator of cAMP production） was also augmented by Nef in a dose-dependent manner （P 〈 0.05）. The basal concentration of cGMP in corpus cavernosum tissue is 0.44 ± 0.09 pmol/mg. Nef did not affect this concentration of cGMP, either in the presence or in the absence of a guanyl cyclase inhibitor （1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, ODQ） （P 〉 0.05）. Also, sodium nitroprusside （SNP, a stimulator of cGMP production）-induced cGMP production was not enhanced by Nef （P 〉 0.05）. Conclusion： Nef, with its relaxation mechanism, can enhance the concentration of cAMP in rabbit corpus cavernosum tissue, probably by inhibiting phosphodiesterase activity. （Asian JAndro12008 Mar; 10： 307-312）

Low-power laser irradiation promotes the proliferation and osteogenic differentiation of human periodontal ligament cells via cyclic adenosine monophosphate

Retaining or improving periodontal ligament （PDL） function is crucial for restoring periodontal defects. The aim of this study was to evaluate the physiological effects of low-power laser irradiation （LPLI） on the proliferation and osteogenic differentiation of human PDL （hPDL） cells. Cultured hPDL cel Is were irradiated （660 nm） daily with doses of O, 1, 2 or 4 J .cm-2. Cell proliferation was evaluated by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide （MTT） assay, and the effect of LPLI on osteogenic differentiation was assessed by Alizarin Red S staining and alkaline phosphatase （ALP） activity. Additionally, osteogenic marker gene expression was confirmed by real-time reverse transcription-polymerase chain reaction （RT-PCR）. Our data showed that LPLI at a dose of 2 J.cm-2 significantly promoted hPDL cell proliferation at days 3 and 5. In addition, LPLI at energy doses of 2 and 4 J.cm-2 showed potential osteogenic capacity, as it stimulated ALP activity, calcium deposition, and osteogenic gene expression. We also showed that cyclic adenosine monophosphate （cAMP） is a critical regulator of the LPLI-mediated effects on hPDL cells. This study shows that LPLI can promote the proliferation and osteogenic differentiation of hPDL cells. These results suggest the potential use of LPLI in clinical applications for periodontal tissue regeneration.

Icariin upregulates phosphorylated cyclic adenosine monophosphate response element binding protein levels in the hippocampus of the senescence-accelerated mouse

At 8 weeks after intragastric administration of icariin to senescence-accelerated mice （P8 strain）, Morris water maze results showed that escape latency was shortened, and the number of platform crossings was increased. Immunohistochemical staining and western blot assay detected significantly increased levels of cyclic adenosine monophosphate response element binding protein These results suggest that icariin upregulates phosphorylated cyclic adenosine monophosphate response element binding protein levels and improves learning and memory functions in hippocampus of the senescence-accelerated mouse.

Analysis of the nitric oxide-cyclic guanosine monophosphate pathway in experimental liver cirrhosis suggests phosphodiesterase-5 as potential target to treat portal hypertension

AIM To investigate the potential effect of inhibitors of phosphodiesterase-5(PDE-5) for therapy of portal hypertension in liver cirrhosis.METHODS In the rat model of thioacetamide-induced liver fibrosis/cirrhosis the nitric oxide-cyclic guanosine monophosphate(NO-cGMP) pathway was investigated. Expression and localization of PDE-5, the enzyme that converts vasodilating cGMP into inactive 5'-GMP, was in the focus of the study. Hepatic gene expression of key components of the NO-cGMP pathway was determined by qRT-PCR: Endothelial NO synthase(eNOS), inducible NO synthase(iNOS), soluble guanylate cyclase subunits α1 and β1(sGCa1, sGCb1), and PDE-5. Hepatic PDE-5 protein expression and localization were detected by immunohistochemistry. Serum cGMP concentrations were measured using ELISA. Acute effects of the PDE-5 inhibitor Sildenafil(0.1 mg/kg or 1.0 mg/kg) on portal and systemic hemodynamics were investigated using pressure transducers.RESULTS Hepatic gene expression of eNOS(2.2-fold; P = 0.003), sGCa1(1.7-fold; P = 0.003), sGCb1(3.0-fold; P = 0.003), and PDE-5(11-fold; P = 0.003) was increased in cirrhotic livers compared to healthy livers. Overexpression of PDE-5(7.7-fold; P = 0.006) was less pronounced in fibrotic livers. iNOS expression was only detected in fibrotic and cirrhotic livers. In healthy liver, PDE-5 protein was localized primarily in zone 3 hepatocytes and to a lesser extent in perisinusoidal cells. This zonation was disturbed in cirrhosis: PDE-5 protein expression in perisinusoidal cells was induced approximately 8-fold. In addition, PDE-5-expressing cells were also found in fibrous septa. Serum cGMP concentrations were reduced in rats with cirrhotic livers by approximately 40%. Inhibition of PDE-5 by Sildenafil caused a significant increase in serum cGMP concentrations [+ 64% in healthy rats(P = 0.024), + 85% in cirrhotic rats(P = 0.018)]. Concomitantly, the portal venous pressure was reduced by 19% in rats with liver cirrhosis. CONCLUSION Overexpression and abrogated zonation of PDE-5 likely contribute to the pathogenesis of cirrhotic portal hypertension. PDE-5 inhibition may therefore be a reasonable therapeutic approach for portal hypertension.

Existence of Heme Oxygenase-carbon Monoxide-cyclic Guanosine Monophosphate Pathway in Human Trabecular Meshwork Cells In Vitro

To confirm the existence of heme oxygenase (HO)-carbon monoxide (CO)- cyclic guanosine monophosphate (cGMP) pathway in the cultured human trabecular meshwork cells (HTMCs) in vitro, and to evaluate the inductive role of hemin on this pathway, HTMCs of the third to fourth generation were cultured in vitro. Reverse transcripase-polymerase chain reaction (RT-PCR) was employed for detection of HO-1 and HO-2 mRNA. Immunohistochemical staining was used to detect HO-1 and HO-2 proteins. Hemin was added into the culture solution. The HO-1 mRNA levels were quantified by RT-PCR. The relative amount of carbon monoxide released into the media was measured with the quantifying carbon monoxide hemoglobin (HbCO) by spectrophotometry. Radioimmunoassay was used to determine changes of cGMP in HTMCs. The results showed that cultured cells had the specific characteristics of HTMCs. Both HO-1 and HO-2 genes were expressed in HTMCs, as well as HO-1 and HO-2 proteins in HTMCs. Hemin induced HO-1 mRNA, HbCO and cGMP in a dose-dependent manner. In conclusion, HO-CO-cGMP pathway exists in the cultured HTMCs and can be induced by hemin. Pharmacological stimulation of HO-CO-cGMP pathway may constitute a novel therapeutic approach to rescuing glaucoma.

Efficient production of cyclic adenosine monophosphate from adenosine triphosphate by the N-terminal half of adenylate cyclase from Escherichia coli

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.

Diabetes and inflammatory diseases:An overview from the perspective of Ca^(2+)/3'-5'-cyclic adenosine monophosphate signaling

A large amount of evidence has supported a clinical link between diabetes and inflammatory diseases,e.g.,cancer,dementia,and hypertension.In addition,it is also suggested that dysregulations related to Ca^(2+)signaling could link these diseases,in addition to 3'-5'-cyclic adenosine monophosphate(cAMP)signaling pathways.Thus,revealing this interplay between diabetes and inflammatory diseases may provide novel insights into the pathogenesis of these diseases.Publications involving signaling pathways related to Ca^(2+)and cAMP,inflammation,diabetes,dementia,cancer,and hypertension(alone or combined)were collected by searching PubMed and EMBASE.Both signaling pathways,Ca^(2+)and cAMP signaling,control the release of neurotransmitters and hormones,in addition to neurodegeneration,and tumor growth.Furthermore,there is a clear relationship between Ca^(2+)signaling,e.g.,increased Ca^(2+)signals,and inflammatory responses.cAMP also regulates pro-and anti-inflammatory responses.Due to the experience of our group in this field,this article discusses the role of Ca^(2+)and cAMP signaling in the correlation between diabetes and inflammatory diseases,including its pharmacological implications.As a novelty,this article also includes:(1)A timeline of the major events in Ca^(2+)/cAMP signaling;and(2)As coronavirus disease 2019(COVID-19)is an emerging and rapidly evolving situation,this article also discusses recent reports on the role of Ca^(2+)channel blockers for preventing Ca^(2+)signaling disruption due to COVID-19,including the correlation between COVID-19 and diabetes.

ADSL, AMPD1, and ATIC Expression Levels in Muscle and Their Correlations with Muscle Inosine Monophosphate Content in Dapulian and Hybridized Pig Species

We investigated the relationship between muscle inosine monophosphate (IMP) content and mRNA levels of ADSL, AMPD1, and ATIC in Dapulian (DPL), Landrace × Dapulian (LDPL), and Duroc × Landrace × Dapulian (DLDPL) hybridized pigs. Methods: The total RNA in longissimus dorsi was isolated from Dapulian (DPL), Landrace × Dapulian (LDPL) and Duroc × Landrace × Dapulian (DLDPL) hybridized pigs, weighed about 95.0 kg, n = 8/species. The internal genes with highest stability (YWHAZ and RPL4) were chosen from 11 common internal genes using Quantitative real-time PCR (qPCR) and geNorm software. The mRNA levels of ADSL, AMPD1 and ATIC genes were corrected with YWHAZ and RPL4 genes. The muscular IMP content was determined by HPLC. The muscular IMP content in DPL was higher than that in LDPL and DLDPL, 25.00% (p 0.05) and 15.56% (p > 0.05), respectively. The muscular mRNA level of ADSL gene in DPL and LDPL was higher than that in DLDPL, 24.14% and 12.07%, respectively (p 0.05). The muscular mRNA level of ATIC gene in DPL and LDPL was higher than that in DLDPL, 66.67% and 33.33%, respectively (p 0.05). The muscular mRNA level of AMPD1 gene in DPL and LDPL was higher than that in DLDPL, 14.49% and 33.26%, respectively. Furthermore, the IMP content was positively correlated with the mRNA level of ADSL, AMPD1 and ATIC genes, respectively (p 0.05). The mRNA level of ADSL gene was highly related to that of AMPD1 and ATIC gene, respectively (p 0.01), while that of AMPD1 gene was not strongly correlated with that of ATIC gene (p > 0.05). The muscular mRNA level of AMPD1, ADSL and ATIC genes and the muscular IMP content in DPL were highest, followed by those in LDPL and DLDPL. The muscular IMP content was positively correlated with the muscular mRNA level of ADSL, AMPD1 and ATIC genes, respectively.

Symptomatic improvement in an acute, non-traumatic spine pain model with a combination of uridine triphosphate, cytidine monophosphate, and hydroxocobalamin

Rationale: In a previously published trial on spinal acute non-traumatic pain, peripheral neuro- regenerative combination of UTP, CMP and hydroxocobalamin presented unexpected analgesicproperties. Objective: To corroborate analgesiceffects of UTP, CMP and hydroxocobalamin combination in a self-paired evolutionary model. Methods: Mean VAS scores from pretreatment, V2 (5th treatment day) and V3 (10th treatment day) were plotted and statistically analyzed (ANOVA) for differences. PFQ scores from pretreatment, V2, and V3 were analyzed using the chisquare test. Results: The difference between V3 and pretreatment mean VAS scores was statistically significant (p < 0.0001). The improvement in PFQ scores throughout the study was found to be statistically significant (p < 0.0001). Conclusion: The combination of UTP, CMP and hydroxocobalamin seems to have analgesic properties in mediumterm use. The complex peripheral neu-roregenerative pharmacodynamics of this combination provides a plausible basis for this finding. Further randomized studies are needed to explore this combination for the indication of neuropathic pain due to spinal structure involvement.

Cyclic adenosine monophosphate signal pathway is involved in regulation of triacylglycerol biosynthesis following nitrogen deprivation in Chlamydomonas reinhardtii

The unicellular green alga,Chlamydomonas reinhardtii is a model organism for studying various biological processes,such as photosynthesis,flagellar motility,and lipid metabolism.To find some novel genes regulating the lipid metabolism under various stress conditions,the paromomycin resistance gene aphVIII was transferred into the genome of C.reinhardtii to establish a mutant library.Two genes mutated in two of the TAG-reduced mutants(Cre06.g278111 in M2 mutant,Cre06.g278110 in M6 mutants)were neighboring in the genome,and their expression levels were down-regulated in their corresponding mutants in parallel with their reduced TAG levels following N deprivation.The proteins encoded by these two genes(KCN11 by Cre06.g278111,ACYC3 by Cre06.g278110)contained a conversed cyclic mononucleotide phosphate(cNMP)binding protein and an adenylate domain,respectively.Since cNMP binding protein and adenylate domain have been known as important components of cyclic adenosine monophosphate(cAMP)signaling pathway,suggesting that these two genes might af fect cellular TAG biosynthesis through cAMP signal pathway.

Sevoflurane effects on cyclic adenosine monophosphate response element binding protein,phosphorylated cyclic adenosine monophosphate response element binding protein,and Livin expression in the cortex and hippocampus of a vascular cognitive impairment rat

BACKGROUND： Neuronal necrosis and apoptosis play important roles in the pathophysiology of cerebral ischemia and resulting cognitive impairment. However, inhibition of neuronal necrosis and apoptosis has been shown to attenuate cognitive impairment following cerebral ischemia. OBJECTIVE： To investigate the effects of sevoflurane on cyclic adenosine monophosphate response element binding protein （CREB）, phosphorylated CREB （pCREB）, and Livin expression in the cortex and hippocampus of a rat model of vascular cognitive impairment.DESIGN, TIME AND SETTING： A randomized, controlled experiment was performed in the Chongqing Key Laboratory of Neurology between June 2007 and July 2008.MATERIALS： Sevoflurane was provided by Abbott Laboratory, UK; Morris water maze was provided by Chinese Academy of Medical Sciences, China; goat anti-rat CREB, goat anti-rat pCREB and goat anti-rat Livin antibodies were provided by Biosource International, USA. METHODS： A total of 42 female, Wistar rats were randomly assigned to the following groups： sham operation, vascular cognitive impairment, and sevoflurane treatment. The vascular cognitive impairment rat model was established by permanent bilateral occlusion of both common carotid arteries, and 1.0 MAC sevoflurane was immediately administered by inhalation for 2 hours. MAIN OUTCOME MEASURES： CREB, pCREB, and Livin expression was measured in the cortex and hippocampus by Western blot and reverse transcription-polymerase chain reaction. Behavior was evaluated with Morris water maze. RESULTS： CREB, pCREB, and Livin expression in the sevoflurane treatment group was significantly greater than the vascular cognitive impairment group （P 〈 0.01）. However, expression of CREB and pCREB was significantly less in the sevoflurane treatment and vascular cognitive impairment groups, compared with the sham operation group （P 〈 0.01）. Livin expression in the sevoflurane treatment and vascular cognitive impairment groups was significantly greater than the sham operation group （P 〈 0.01）. Learning, memory, and behavior disorders were observed in the vascular cognitive impairment group. Sevoflurane treatment significantly improved these observed disorders. CONCLUSION： Sevoflurane improved cognitive impairment due to permanent bilateral occlusion of both common carotid arteries. Improved function was associated with increased CREB, pCREB, and Livin expression in the cortex and hippocampus.

Excess-Electron Attachment and Ionization of Aqueous Uridine Monophosphate Anion

We applied quantum mechanics/classical mechanics simulations to study excess-electron attachment and ionization of uridine monophosphate anion(d UMP-)in explicit aqueous solutions.We calculated vertical electron affinities(VEAs),adiabatic electron affinities(AEAs),vertical detachment energies(VDEs),vertical ionization energies(VIEs),and adiabatic ionization energies(AIEs)of the 40 structures obtained from molecular dynamic trajectory.The excess-electron and hole distributions were analyzed in electron attachment and ionization of aqueous d UMP^(-).The converged mean VEA(-0.31 e V)and AEA(2.13 e V)suggest that excess-electron can easily attach to d UMP^(-).The mean vertical(-0.50 e)and adiabatic(-0.62 e)excess-electron on uracil reveal that main excesselectrons are localized on nucleobases at the most snapshots.The distributions at several special snapshots demonstrate the excess-electron delocalization over nucleobases/ribose or ribose/phosphate group after the structural relaxations of d UMP^(2-)dianion.The VDE value(2.78 e V)indicates that d UMP2-dianion could be very stable.Moreover,the mean VIE is 8.13 eV which is in agreement with the previous calculation using solvation model.The hole distributions on uracil suggest that the nucleobases are easily ionized after the irradiation of high-energy rays.In vertical ionizations,the holes would be delocalized over uracil and ribose at several snapshots.Observing the adiabatic hole distributions,it can be found that electrons on phosphate group and holes on nucleobases can be transferred to ribose at the special snapshots in the structural relaxation of neutral species.

Absorption of Curcumin Monophosphate in Rat Intestinal Circulation

[Objectives] This study aimed to study the absorption of curcumin monophosphate, a derivative of curcumin, in the small intestine of rats. [Methods] In situ recirculation perfusion technique was used to study the absorption of curcumin monophosphate in intestinal circulation of rats, and the content of curcumin monophosphate in intestinal circulation solution was determined by high-performance liquid chromatography/ultraviolet detector(HPLC/UV). [Results] The established HPLC/UV method has good specificity. Linear regression was conducted between the peak area of curcumin monophosphate(A) and the concentration of curcumin monophosphate(C). The established standard curve equation was y=30.07x+102.48(R^2=0.999 0), indicating that curcumin monophosphate has good linearity in the range of 1.25-100.00 μg/mL. After excluding the degradation of the drug and the loss of sampling, within the first 2 h of the experiment, the drug absorption in the small intestine of rat was 1.39 mg, accounting for 97.2% of the total drug absorption during the 5-h experimental period, and the absorption rate was 53.9%.[Conclusions] Curcumin monophosphate has a good absorption in the small intestine of rats.

A New Mixed-metal Monophosphate:Ba2Bi2Co(PO4)4

A new member of mixed-metal Ba2Bi2M-Ⅱ（PO4）4 monophosphate, namely Ba2Bi2Co（PO4）4, was synthesized by solid state method and characterized by X-ray single-crystal diffraction and powder diffraction for the first time. It crystallizes in the orthorhombic system with space group Pnma（No. 62） and features a 3D architecture built up of adjacent zig-zag linear structures of [CoP4O（16）]∞ along [100], and further connected by [Bi2O（11）] dimers to form a 3D framework, where the Ba2＋ are located in the free space. The stereochemical activity of the Bi3＋ lone pair has also been discussed. The result of magnetic property measurement confirms the antiferromagnetic property of Ba2Bi2Co（PO4）4.

Studies on Relationship between Serum Nitric Oxide and Plasma Cyclic Guanosine Monophosphate and Prolonged Bleeding after Medical Abortion as well as Prophylaxis and Treatment of Bleeding with Traditional Chinese Medicine

Objectives To study the relationship between serum nitric oxide (NO and plasma cyclic guanosine monophosphate (cGMP) and prolonged bleeding after medical abortion. Methods A total of 120 women having received medical abortions at random were recruited and divided into two groups: the one (Group A,n=60) taking 'Gong Fu Mixture(Uterus Recovering Mixture)' and the other (Group B,n=60) not taking it after abortion. On d 10, 20 and 30 after medical abortion, serum NO and plasma cGMP were tested before and after mifepristone administration and 10 d later by Gresis reaction method and radioimmunoassay respectively. Results NO concentration in serum and cGMP concentration in plasma decreased significantly after taking mifepristone given (P<0.05). Ten days later, the number of those with bleeding discontinuation in the group A was significantly greater than that in the group B (P<0.05). Serum NO level and plasma cGMP level in the group A decreased more significantly than those in the group B (P<0.05). Conclusion The slow decrease of serum NO and plasma cGMP is closely related to prolonged bleeding after medical abortion. “Gong Fu Mixture (uterus recovering mixture)” is effective in prevention and treatment of prolonged bleeding.

Inhibition of matrix metalloproteinase-9 secretion by dimethyl sulfoxide and cyclic adenosine monophosphate in human monocytes

BACKGROUND Matrix metalloproteinases(MMPs),including MMP-9,are an integral part of the immune response and are upregulated in response to a variety of stimuli.New details continue to emerge concerning the mechanistic and regulatory pathways that mediate MMP-9 secretion.There is significant evidence for regulation of inflammation by dimethyl sulfoxide(DMSO)and 3',5'-cyclic adenosine monophosphate(cAMP),thus investigation of how these two molecules may regulate both MMP-9 and tumor necrosis factorα(TNFα)secretion by human monocytes was of high interest.The hypothesis tested in this study was that DMSO and cAMP regulate MMP-9 and TNFαsecretion by distinct mechanisms.AIM To investigate the regulation of lipopolysaccharide(LPS)-stimulated MMP-9 and tumor necrosis factorαsecretion in THP-1 human monocytes by dimethyl sulfoxide and cAMP.METHODS The paper describes a basic research study using THP-1 human monocyte cells.All experiments were conducted at the University of Missouri-St.Louis in the Department of Chemistry and Biochemistry.Human monocyte cells were grown,cultured,and prepared for experiments in the University of Missouri-St.Louis Cell Culture Facility as per accepted guidelines.Cells were treated with LPS for selected exposure times and the conditioned medium was collected for analysis of MMP-9 and TNFαproduction.Inhibitors including DMSO,cAMP regulators,and anti-TNFαantibody were added to the cells prior to LPS treatment.MMP-9 secretion was analyzed by gel electrophoresis/western blot and quantitated by ImageJ software.TNFαsecretion was analyzed by enzyme-linked immuno sorbent assay.All data is presented as the average and standard error for at least 3 trials.Statistical analysis was done using a two-tailed paired Student t-test.P values less than 0.05 were considered significant and designated as such in the Figures.LPS and cAMP regulators were from Sigma-Aldrich,MMP-9 standard and antibody and TNFαantibodies were from R&D Systems,and amyloid-βpeptide was from rPeptide.RESULTS In our investigation of MMP-9 secretion from THP-1 human monocytes,we made the following findings.Inclusion of DMSO in the cell treatment inhibited LPSinduced MMP-9,but not TNFα,secretion.Inclusion of DMSO in the cell treatment at different concentrations inhibited LPS-induced MMP-9 secretion in a dosedependent fashion.A cell-permeable cAMP analog,dibutyryl cAMP,inhibited both LPS-induced MMP-9 and TNFαsecretion.Pretreatment of the cells with the adenylyl cyclase activator forskolin inhibited LPS-induced MMP-9 and TNFαsecretion.Pretreatment of the cells with the general cAMP phosphodiesterase inhibitor IBMX reduced LPS-induced MMP-9 and TNFαin a dose-dependent fashion.Pre-treatment of monocytes with an anti-TNFαantibody blocked LPSinduced MMP-9 and TNFαsecretion.Amyloid-βpeptide induced MMP-9 secretion,which occurred much later than TNFαsecretion.The latter two findings strongly suggested an upstream role for TNFαin mediating LPS-stimulate MMP-9 secretion.CONCLUSION The cumulative data indicated that MMP-9 secretion was a distinct process from TNFαsecretion and occurred downstream.First,DMSO inhibited MMP-9,but not TNFα,suggesting that the MMP-9 secretion process was selectively altered.Second,cAMP inhibited both MMP-9 and TNFαwith a similar potency,but at different monocyte cell exposure time points.The pattern of cAMP inhibition for these two molecules suggested that MMP-9 secretion lies downstream of TNFαand that TNFαmay a key component of the pathway leading to MMP-9 secretion.This temporal relationship fit a model whereby early TNFαsecretion directly led to later MMP-9 secretion.Lastly,antibody-blocking of TNFαdiminished MMP-9 secretion,suggesting a direct link between TNFαsecretion and MMP-9 secretion.

The cGAS-STING-interferon regulatory factor 7 pathway regulates neuroinflammation in Parkinson's disease

Interferon regulatory factor 7 plays a crucial role in the innate immune response.However,whether interferon regulatory factor 7-mediated signaling contributes to Parkinson's disease remains unknown.Here we report that interferon regulatory factor 7 is markedly up-regulated in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced mouse model of Parkinson's disease and co-localizes with microglial cells.Both the selective cyclic guanosine monophosphate adenosine monophosphate synthase inhibitor RU.521 and the stimulator of interferon genes inhibitor H151 effectively suppressed interferon regulatory factor 7 activation in BV2 microglia exposed to 1-methyl-4-phenylpyridinium and inhibited transformation of mouse BV2 microglia into the neurotoxic M1 phenotype.In addition,si RNA-mediated knockdown of interferon regulatory factor 7 expression in BV2 microglia reduced the expression of inducible nitric oxide synthase,tumor necrosis factorα,CD16,CD32,and CD86 and increased the expression of the anti-inflammatory markers ARG1 and YM1.Taken together,our findings indicate that the cyclic guanosine monophosphate adenosine monophosphate synthase-stimulator of interferon genes-interferon regulatory factor 7 pathway plays a crucial role in the pathogenesis of Parkinson's disease.