Objective To investigate the change of protein expression of lung tissue of rabbit after ischemic preconditioning(IP)and try to elucidate the potential protective mechanism of IP.Methods 12 domestic rabbits were rando...Objective To investigate the change of protein expression of lung tissue of rabbit after ischemic preconditioning(IP)and try to elucidate the potential protective mechanism of IP.Methods 12 domestic rabbits were randomly divided into group IP and group control(6 rabbits in each group).All the left lungs were afflicted by ische mia-reperfusion injury except that those in group IP were subject to IP prior to ischemic phase.2-DE was employed to separate the total protein of the lung tissue.PDQuest analysis software was used to distinguish the differently expressed protein spot.MALDI-TOF-MS and Mascot database searching were exploited to identify these proteins.Results 1)IP attenuated the ischemia-reperfusion lung injury.2)The proteomic analysis showed 35 target proteins,of which 17 were characterized such as phosphatidylinositol 3-kinase(PI3k)delta catalytic subunit.Conclusions 1)Proteomic is a promising tool to investigate the IP and ischemia-reperfusion lung injury.2)That IP inhibits inflammatory cascades through phosphatidylinositol 3-kinase signal transduction pathway may be one of its protective mechanism.展开更多
OBJECTIVE In this study we explored the role of Epac1-Rap1 pathway in the acute myocardial ischemia/reperfusion injury(MIRI) in vitro and in vivo.METHODS An acute myocardial ischemia/reperfusion injury model was estab...OBJECTIVE In this study we explored the role of Epac1-Rap1 pathway in the acute myocardial ischemia/reperfusion injury(MIRI) in vitro and in vivo.METHODS An acute myocardial ischemia/reperfusion injury model was established by the ligation of left anterior descending coronary.Myocardial architecture,fibers and apoptosis was evaluated by the Masson trichrome staining,Sirius red staining and TUNEL assay.H9c2 cells were subjected to hypoxia for 5 h followed by 1-h reoxygen.ation in vitro.Cell viability was measured by MTT assay and cellular injury was evaluated by measuring the release of lactate dehydrogenase(LDH).Western blot,real-time PCR and immunofluorescence were used to detect the expressions of Epac1 and relative downstream molecules.RESULTS Myocardial IR-induced cardiac apoptosis and accumulation of Epac1 and Rap1 in rat IR injury model.Direct Epac activation by 8-CPT(8-(4-chlorophenylthio)-2′-O-methyl-cAMP) exacerbated cardiomyocyte death and dysfunction following hypoxia-reoxygenation(H/R),selective activation of Epac in response to H/R was evident which enriched for cytosolic/membrane proteins and mRNA.Harmacological inhibitor of Epac(ESI-09) significantly ameliorated myocardial injury with the decline of Epac expression.Epac inhibitor and agonist studies also implicated the effect of Rap1,which is downstream of Epac in this pathway.The expression of Rap1 elevated when activated by Epac agonist and was blocked by Epac inhibitor.The same result was true for myocyte CaMK-II and intracellular calcium ions activation.Moreover,ESI-09 also increased ERK1/2 phosphorylation.CONCLUSION Our study reveal that Epac1/Rap1 signaling pathway is involved in the pathogenesis of myocardial I/R injury in rats,which provides evidence on the development of therapeutic strategies target this pathway for myocardial I/R injury.展开更多
OBJECTIVE The total steroid saponins(TSSN)isolated from Dioscorea zingiberensis C.H.Wright(D.zingiberensis)has shown a variety of beneficial bioactivities.However,there are no reports about the neuroprotective effects...OBJECTIVE The total steroid saponins(TSSN)isolated from Dioscorea zingiberensis C.H.Wright(D.zingiberensis)has shown a variety of beneficial bioactivities.However,there are no reports about the neuroprotective effects of the TSSN until now.Therefore,we explored the neuroprotective effects of TSSN on rats against transient focal cerebral ischemia-reperfusion(I/R)and the underlying mechanisms.METHODS The healthy adult Sprague-Dawley rats were randomly assigned into six groups.After pre-treatment with the TSSN intragastrically for six days,the rats were subjected to the ischemia injury by the surgery of middle cerebral artery occlusion(MCAO)for 90 min.Some indexes were evaluated and detected.RESULTS As compared to the I/R group,TSSN group of rats,especially given the 30 mg·kg^-1 of TSSN,not only marked reduction in the neurological deficit scores,cerebral infarct volume,and brain edema,but also an increase in neuron survival(Nissl bodies)in the hippocampal cornuammons 1(CA1)and cortex hemisphere of the ipsilateral ischemia.At the same time,the inflammatory cytokines in serum induced by MCAO were significantly alleviated by the TSSN pre-administration.What′s more,the increase of caspase-3 was evidently reduced in the CA1 and cortex of the hemisphere injured brain.Finally,the down-regulating anti-apoptotic Bcl-2 and up-regulating pro-apoptotic Bax proteins were obviously suppressed.CONCLUSION TSSN plays a potential neuroprotective role against a severe injury induced by transient focal cerebral ischemic reperfusion in a rat experimental model,and this role may be mediated by its antiinflammatory and anti-apoptotic actions.展开更多
OBJECTIVE To predict the potential targets of hyperoside(Hyp)on improving ischemia/reperfusion injury by network pharmacology,and explore its possible mechanism combined with related literature.METHODS The action targ...OBJECTIVE To predict the potential targets of hyperoside(Hyp)on improving ischemia/reperfusion injury by network pharmacology,and explore its possible mechanism combined with related literature.METHODS The action targets of Hyp and ischemia/reperfusion injury were obtained by TCMSP,Swiss Target Prediction,Pharm Mapper,Similarity ensemble approach,Online Mendelian Inheritance in Man,DisGENT and database.The common targets of drugs and diseases were screened by Omishare and STRING database respectively,and the protein-protein interaction(PPI)network map was constructed.Then the interaction network between Hyp and disease targets was constructed by Cytoscape software and topological cross-linking analysis was carried out.Then the interaction network between Hyp and disease targets was constructed and cross-linked analysis was carried out by using Cytoscape software.The gene ontology(GO)of the core target was analyzed by David database,and then the related pathways of the core target were enriched by KEGG database.RESULTS A total of 54 GO enrichment processes were obtained by GO enrichment analysis of 44 common genes,including 38 biological processes(BP),15 cell composition(CC)processes,and 1 molecular functional(MF)process.43 items were obtained by signal pathway enrichment analysis in KEGG database.CONCLUSION It is suggested that the mechanism of Hyp may be related to PI3K-Akt,RAP1,RAS,VEGF and other signal transduction pathways.The above results laid a theoretical foundation for the study of the mechanism and clinical application of the treatment of ischemia/reperfusion injury.展开更多
The morbidity and mortality of cardiovascular diseases are very high,which has attracted more and more attention all over the world.Common treatment methods for clinical treatment of acute myocardial infarction includ...The morbidity and mortality of cardiovascular diseases are very high,which has attracted more and more attention all over the world.Common treatment methods for clinical treatment of acute myocardial infarction include direct percutaneous coronary intervention and coronary artery bypass grafting,which can quickly restore blocked coronary blood flow and reduce the infarct size.However,the inevitable ischemia/reperfusion injury will occur during the recovery of coronary blood flow,its pathological mechanism is complicated,and the Western medicine countermeasures are very limited.Among the current drugs for the treatment of cardiovascular diseases,traditional Chinese medicine has become a research hotspot due to its multiple targets,safety,and low side effects.Ginger is the fresh rhizome of Zingiber officinale Rosc.,a perennial herbaceous plant in the ginger family.It is a dual-purpose resource of medicine and food.Ginger has the functions of relieving the appearance and dispelling cold,warming up and relieving vomiting,resolving phlegm and relieving cough,and relieving fish and crab poison.The chemical components of ginger mainly include volatile oil,gingerol,diphenylheptane,etc..Among them,6-gingerol,as the main active component of gingerols,has obvious pharmacological effects in myocardial protection,anti-oxidation,anti-inflammatory,etc..Studies have shown that 6-gingerol protects myocardium mainly through anti-oxidative stress,anti-inflammatory,inhibiting cell apoptosis,and preventing calcium influx.①Anti-oxidative stress:oxidative stress is a state where oxidation and anti-oxidation in the body are out of balance,and it is also an important factor leading to myocardial damage.Many studies have confirmed that 6-gingerol has an antioxidant effect,and it is considered a natural antioxidant.6-gingerol can significantly reduce the degree of oxidative stress and the level of reactive oxygen species caused by cardiomyocyte damage,and has a significant cardioprotective effect.②Anti-inflammatory:inflammation can cause substantial cell damage and organ dysfunction,which is another important cause of myocardial damage.6-gingerol can reduce the levels of inflammatory factors such as interleukin-6,interleukin-1β,and tumor necrosis factor-αin cardiomyocytes,and at the same time inhibit the TLR4/NF-κB signaling pathway,an important regulatory pathway of inflammation,showing that it may improve myocardial damage through anti-inflammatory effects.③Inhibition of apoptosis:apoptosis is a complex and orderly process in the autonomous biochemical process of cells,and one of the main mechanisms of myocardial injury.This process can be roughly divided into three pathways:mitochondria,endoplasmic reticulum,and death receptors.Among them,the mitochondrial pathway plays an important role,and Bcl-2 and Bax located upstream of this pathway can regulate the entire process of cell apoptosis by regulating the permeability of the mitochondrial membrane.Studies have found that the preventive application of 6-gingerol can reduce cell damage,reduce the number of apoptotic cells,reduce the activity of Bax and caspase-3,and increase the expression of Bcl-2.Therefore,6-gingerol pretreatment can reduce the damage of cardiomyocytes,and its mechanism may be related to the inhibition of apoptosis.④Prevent calcium influx:calcium overload is involved in the pathogenesis of myocardial ischemic injury,which may be related to excessive contracture,arrhythmia,and mitochondrial Ca2+accumulation that impairs myocardial function.6-gingerol inhibits the increase of intracellular Ca2+concentration by inhibiting L-type calcium current,thereby reducing extracellular Ca2+influx,thereby avoiding calcium overload and playing a cardioprotective effect.In summary,6-gingerol can effectively treat and improve myocardial ischemia/reperfusion injury,and it has great development potential in the fields of medicine and health products.展开更多
OBJECTIVE To explore the effect of total flavonoids of Rhododendra simsii(TFR)on improving cerebral ischemia/reperfusion injury(CIRI)and its relationship with STIM/Orai-regulated operational Ca^(2+)influx(SOCE)pathway...OBJECTIVE To explore the effect of total flavonoids of Rhododendra simsii(TFR)on improving cerebral ischemia/reperfusion injury(CIRI)and its relationship with STIM/Orai-regulated operational Ca^(2+)influx(SOCE)pathway.METHODS Oxygen-glucose deprivation/reoxygenation(OGD/R)PC12 cells were used to simulate CIRI in vitro,and the intracellular Ca^(2+)concentration and apoptosis rate of PC12 cells were detected by laser confocal microscope and flow cytometry,respectively.The regulation of STIM/Orai on SOCE was analyzed by STIM/Orai gene silencing and STIM/O rai gene overexpression.The CIRI model was established by MCAO in SD rats.The activities of inflammatory cytokines IL^(-1),IL-6 and TNF-αin serum were detected by ELISA.The pathological changes of ischemic brain tissue and the infarction of rat brain tissue were detected by HE staining and TTC staining.The protein and mRNA expression levels of STIM1,STIM2,Orai1,caspase-3 and PKB in brain tissue were detected by Western blotting and RT-qPCR,respectively.RESULTS The results of in vitro experiment showed that the fluorescence intensity of Ca^(2+)and apoptosis rate in PC12 cells treated with TFR were significantly lower than those in OGD/R group,and this trend was enhanced by SOCE antagonist 2-APB.STIM1/STIM2/Orai1 gene silencing significantly reduced apoptosis and Ca^(2+)overload in OGD/R model,while TFR combined with overexpression of STIM1/STIM2/Orai1 aggravated apoptosis and Ca2+overload.In the in vivo experiment,TFR significantly reduced the brain histopathological damage,infarction of brain tissue,the contents of IL^(-1),IL-6 and TNF-αin the serum in MCAO rats and down-regulated the expression of STIM1,STIM2,Orai1 and caspase-3 protein and mRNA in the brain tissue,and up-regulated the expression of PKB.The above effects were enhanced by the addition of 2-APB.CONCLUSION The above results indicate that TFR may reduce the contents of inflammatory factors and apoptosis,decrease Ca2+overload and ameliorate brain injury by inhibiting SOCE pathway mediated by STIM and Orai,suggesting that it has a protective effect against subacute CIRI.展开更多
Aim Salvia miltiorrhiza Bunge （SM） and lignum dalbergiae odoriferae （DO） are both traditional Chi- nese medicine that have cardioprotective effects. Here, we further examined the combined effects of SM and DO on r...Aim Salvia miltiorrhiza Bunge （SM） and lignum dalbergiae odoriferae （DO） are both traditional Chi- nese medicine that have cardioprotective effects. Here, we further examined the combined effects of SM and DO on rat myocardial ischemia/reperfusion injury. The possible mechanism of SM and DO also were elucidated. Methods DO was divided into aqueous extract of lignum dalbergiae odoriferae （DOW） and lignum dalbergiae odoriferae oil （DOO）. Sprague-Dawley rats were randomized to seven groups： sham group, model group, treatment groups inclu- ding SM （10 g · kg^-1）, DOW （5 g · kg^-1）, DOO （0.5 ml · kg^-1）, SM ＋ DOW （10 g · kg^-1 ＋ 5 g · kg^-1）, SM ＋ DOO （ 10 g · kg^-1 ＋ 0. 5 ml · kg^-1）. Rats were pretreated with homologous drug for 7 days and then subjec- ted to 30 rain of ischemia followed by 180 rain of reperfusion. Electrocardiogram （ECG） and heart rate were moni- tored and recorded continuously. At the end of reperfusion, blood samples were collected to determine the serum levels of creatine kinase-MB （CK-MB） and lactate dehydrogenase （LDH）. Hearts were harvested to assess heart- body rate, infarct size and histopathological changes as well. Maximum and minimum effective points were deter- mined by measuring indicators associate with myocardial injury at different time-points of reperfusion （Smin, 15min, 30min, 45rain, 60min, 120min, 180min）. The potential therapeutic mechanism of SM and SM ＋ DOO were carried out by detecting superoxide dismutase （SOD）, malondialdehyde （MDA）, tumor necrosis factor-alpha （TNF-alpha） and interleukin 6 （IL-6）. Results The results showed SM and DO can ameliorate cardiac function respectively, and this cardioprotective effect was further strengthened by their combinations. Among all the combi- nations, SM ＋ DOO showed predominant potential to improve ECG and heart rate, reduce heart-body rate （28.5% ＋ 1.4% , P 〈 0.01 vs model） and myocardial infarct size （ 20.96% ＋ 1.61% , P 〈 0.01 vs model, P 〈 0.05 vs SM） , attenuate histopathological damage, decrease the levels of CK-MB and LDH （P 〈 0.01 vs model, P 〈 0.05 vs SM）. The maximum effective points of SM and SM ＋ DOO were 15min and 30rain respectively, and the minimum effective points of them were 180rain. In reducing serum level of MDA, TNF-alpha, IL-6 and increasing SOD activ- ity, SM ＋ DOO was similar to SM. Conclusion The results of this study indicated that SM ＋ DOO have combined effects that are highly effective than single pretreatment against myocardial ischemie reperfusion injury in rats. The possible mechanism of SM and DO were likely through its anti-oxidant and anti-inflammatory properties, and thus may be an effective and promising medicine for both prophylaxis and treatment of ischemic heart disease.展开更多
OBJECTIVE To investigate regulatory effects of hyperoside(Hyp)on IP3/PKC/TRPV4 pathway in rat cerebral basilar artery(CBA)subjected to global cerebral ischemia-reperfusion(I/R).METHODS The model of global cerebral I/R...OBJECTIVE To investigate regulatory effects of hyperoside(Hyp)on IP3/PKC/TRPV4 pathway in rat cerebral basilar artery(CBA)subjected to global cerebral ischemia-reperfusion(I/R).METHODS The model of global cerebral I/R in rats was established by four-vessel occlusions methods.The treated rats were administrated with Hyp(50 mg·kg^-1)group,Hyp(50 mg·kg^-1)+HC-067047(10 mg·kg^-1),Hyp(50 mg·kg^-1)+2APB(2 mg·kg^-1),Hyp(50 mg·kg^-1)+BisI(2.5 mg·kg^-1),Hyp(50 mg·kg^-1)+2APB(2 mg·kg^-1)+BisI(2.5 mg·kg^-1).Hematoxylin-eosin(HE)and Nissl staining were performed and the contents of methane dicarboxylic aldehyde(MDA),neuron-specific enolase(NSE),S100β and the activity of lactic dehydrogenase(LDH)in serum were measured by enzyme-linked immunosorbnent assay(ELISA).The specific blocker N-nitro-L-arginine-methyl-ester(L-NAME)and indomethacin(Indo)were used to delete the prostacyclin(PGI2)and nitric oxide(NO)dependent relaxation.The protein expression level of TRPV4 was detected by Western blotting.Ca2+intensity in vascular smooth muscle cells was measured by confocal laser scanning microscope and flow cytometry was performed to observe the apoptosis of CBA endothelial cells after in vivo administration.RESULTS Hyp induced a dose-dependent relaxation of CBA in IR rats via a PGI2 and NO independent manner,as evidenced by alleviated patho⁃logical changes and up-regulated expression of TRPV4 protein in the endothelial cells from cerebral vessels.Hyp signifi⁃cantly reduced the contents of MDA,NSE,S100βand the activity of LDH in serum and decreased the fluorescence intensity of Ca2+in cerebral vascular smooth muscle cells by in vivo administration.The apoptotic rate of endothelial cells in Hyp treated group was significantly less than that in IR group.CONCLUSION Hyp does in fact ameliorate I/R injury by regulatingIP3/PKC/TRPV4 pathway.展开更多
OBJECTIVE To investigate the prevention and its mechanism of Naojian Tablet,the adjusted prescription from Buyang Huanwu Decoction,during focal ischemic brain injury condition,and study on the main bioactive substance...OBJECTIVE To investigate the prevention and its mechanism of Naojian Tablet,the adjusted prescription from Buyang Huanwu Decoction,during focal ischemic brain injury condition,and study on the main bioactive substance for this function.METHODS To apply the modified middle cerebral artery occlusion(MCAO)model in rats,immunohistochemistry,RT-PCR and Western blotting were applied to determine the expression of CDK5,CDK4/cyclinD1 in the hippocampus tissues in MCAO rat by treating with Naojian Tablet.On the other hand,the processes of metabolism and disposition of several active components in Naojian Tablet and Buyang Huanwu Decoction were analyzed and characterized by using HPLC-Q-TOF method.RESULTS Naojian Tablet(ig,2.41g·kg-1)could significantly decrease the levels of CDK5,CDK4/cyclinD1 in hippocampus tissues in MACO rat after treated by 3d,7d,14 d,28d(P<0.05).The similar mechanism was observed in the Buyang Huanwu Decoction treated MACO rat.Calycosin,fermononetin,senkyunolide and ligustilide in Naojian Tablet was determined simultaneously as the main active ingredient of Naojian Tablet.CONCLUSION The above-mentioned in vivo studies suggested that Naojian Tablet maybe play its prevention effect on nerve cells via its acting on CDK5 signaling and cell cycle pathway,which verified the multi-target and multi active pathways efficiency of tradition Chinese medicine in treating diverse disease.展开更多
Myocardial infarction resulting from coronary atherosclerosis is the leading cause of death in modern soci- ety. Reperfusion is an essential treatment to salvage ischemia myocardium from necrosis, while it also leads ...Myocardial infarction resulting from coronary atherosclerosis is the leading cause of death in modern soci- ety. Reperfusion is an essential treatment to salvage ischemia myocardium from necrosis, while it also leads to addi- tional damage. Therefore, exploring effective medicines to protect the heart from post-ischemic injury is one of the major objectives of cardiovascular research. Berbamine is a nature compound of bisbenzylisochinoline alkaloids from Barberry. We found that it displays positive inotropic and lusitropic effects at lower concentrations by increasing myofilament Ca2＋ sensitivity via a PKCe-dependent signaling pathway. Moreover, berbamine preconditioning con- fers cardioprotection against ischemia/reperfusion （I/R） injury by attenuating the Ca2＋ overloading and preventing the calpain activation through the activating of PI3K-Akt-GSK3β pathway and subsequently opening of the mitoKATP channel. Furthermore, we demonstrate that berbamine postconditioning conferred the cardioprotective effect against I/R injury by the regulation of autophagy. These findings reveal new roles and mechanisms of berbamine in the heart and cardioprotection against I/R injury.展开更多
Aim Reduction of Sheng-Nao-Kang decoction （RSNK）, is a modified traditional Chinese medicinal formula of Sheng-Nao-Kang pill preparation, which is protective in rats against focal cerebral ischemia/reperfusion （I/R...Aim Reduction of Sheng-Nao-Kang decoction （RSNK）, is a modified traditional Chinese medicinal formula of Sheng-Nao-Kang pill preparation, which is protective in rats against focal cerebral ischemia/reperfusion （I/R） injury. In the current study, we investigate the protective effect of RSNK against apoptosis and oxidative damage induced by cerebral I/R and explore the underlying mechanisms. Cerebral I/R injury was induced by in- traluminal middle cerebral artery occlusion （MCAO） for 2 h followed by reperfusion for 24 h in adult male Sprague- Dawley rats. Rats were randomized into seven groups （n- 8）： Sham group, I/R group, RSNK-treated groups （ 0.7 g · kg ^- 1, 1 . 4 g · kg ^- 1 and 2. 8 g · kg^ - 1 ） , nimodipine （NMP） -treated group and Whitmania pigra Whitman （WW）-treated group. Neurological deficit scores, cerebral humidity content and cerebral infarction volume were measured after the 24 h reperfusion. Malondialdehyde （ MDA）, superoxide dismutase （ SOD）, catalase （ CAT）, inducible nitric oxide synthase （iNOS） and total nitric oxide synthase （TNOS） in serum were measured by assay kits for biochemical analysis. Histological structures of the cortex of the ipsilateral ischemic cerebral hemisphere in rats were observed by Nissl staining. The caspase-3 protein content in the hippocampus and cortex was detected by immunohistochemistry. Additionally, Bax and Bcl-2 protein expressions in the injured brain were evaluated by Western blot. RSNK administration not only markedly improved neurological deficit scores, but also reduced cere- bral humidity content and cerebral infarction volume, lowered MDA content, up-regulated SOD and CAT levels, down-regulated iNOS and TNOS levels, restrained the expression of caspase-3 positive protein and alleviated the Bax and Bcl-2 protein expressions.展开更多
基金Project(2004036433)supported by the Postdoctoral Science Foundation of Chinaproject(B2004024)supported by theScience Foundation of Public Health Bureau of Hunan Province
文摘Objective To investigate the change of protein expression of lung tissue of rabbit after ischemic preconditioning(IP)and try to elucidate the potential protective mechanism of IP.Methods 12 domestic rabbits were randomly divided into group IP and group control(6 rabbits in each group).All the left lungs were afflicted by ische mia-reperfusion injury except that those in group IP were subject to IP prior to ischemic phase.2-DE was employed to separate the total protein of the lung tissue.PDQuest analysis software was used to distinguish the differently expressed protein spot.MALDI-TOF-MS and Mascot database searching were exploited to identify these proteins.Results 1)IP attenuated the ischemia-reperfusion lung injury.2)The proteomic analysis showed 35 target proteins,of which 17 were characterized such as phosphatidylinositol 3-kinase(PI3k)delta catalytic subunit.Conclusions 1)Proteomic is a promising tool to investigate the IP and ischemia-reperfusion lung injury.2)That IP inhibits inflammatory cascades through phosphatidylinositol 3-kinase signal transduction pathway may be one of its protective mechanism.
基金This work supported by the National Natural Science Foundation of China (81470432) and Natural Science Foundation of Anhui Province Education Department (KJ2016A357).
文摘OBJECTIVE In this study we explored the role of Epac1-Rap1 pathway in the acute myocardial ischemia/reperfusion injury(MIRI) in vitro and in vivo.METHODS An acute myocardial ischemia/reperfusion injury model was established by the ligation of left anterior descending coronary.Myocardial architecture,fibers and apoptosis was evaluated by the Masson trichrome staining,Sirius red staining and TUNEL assay.H9c2 cells were subjected to hypoxia for 5 h followed by 1-h reoxygen.ation in vitro.Cell viability was measured by MTT assay and cellular injury was evaluated by measuring the release of lactate dehydrogenase(LDH).Western blot,real-time PCR and immunofluorescence were used to detect the expressions of Epac1 and relative downstream molecules.RESULTS Myocardial IR-induced cardiac apoptosis and accumulation of Epac1 and Rap1 in rat IR injury model.Direct Epac activation by 8-CPT(8-(4-chlorophenylthio)-2′-O-methyl-cAMP) exacerbated cardiomyocyte death and dysfunction following hypoxia-reoxygenation(H/R),selective activation of Epac in response to H/R was evident which enriched for cytosolic/membrane proteins and mRNA.Harmacological inhibitor of Epac(ESI-09) significantly ameliorated myocardial injury with the decline of Epac expression.Epac inhibitor and agonist studies also implicated the effect of Rap1,which is downstream of Epac in this pathway.The expression of Rap1 elevated when activated by Epac agonist and was blocked by Epac inhibitor.The same result was true for myocyte CaMK-II and intracellular calcium ions activation.Moreover,ESI-09 also increased ERK1/2 phosphorylation.CONCLUSION Our study reveal that Epac1/Rap1 signaling pathway is involved in the pathogenesis of myocardial I/R injury in rats,which provides evidence on the development of therapeutic strategies target this pathway for myocardial I/R injury.
基金National Natural Science Foundation of China(81603267)Fundamental Research Funds for the Central UniversitiesShaanxi Key Laboratory of "Qiyao"Resources and Anti-tumor Activities
文摘OBJECTIVE The total steroid saponins(TSSN)isolated from Dioscorea zingiberensis C.H.Wright(D.zingiberensis)has shown a variety of beneficial bioactivities.However,there are no reports about the neuroprotective effects of the TSSN until now.Therefore,we explored the neuroprotective effects of TSSN on rats against transient focal cerebral ischemia-reperfusion(I/R)and the underlying mechanisms.METHODS The healthy adult Sprague-Dawley rats were randomly assigned into six groups.After pre-treatment with the TSSN intragastrically for six days,the rats were subjected to the ischemia injury by the surgery of middle cerebral artery occlusion(MCAO)for 90 min.Some indexes were evaluated and detected.RESULTS As compared to the I/R group,TSSN group of rats,especially given the 30 mg·kg^-1 of TSSN,not only marked reduction in the neurological deficit scores,cerebral infarct volume,and brain edema,but also an increase in neuron survival(Nissl bodies)in the hippocampal cornuammons 1(CA1)and cortex hemisphere of the ipsilateral ischemia.At the same time,the inflammatory cytokines in serum induced by MCAO were significantly alleviated by the TSSN pre-administration.What′s more,the increase of caspase-3 was evidently reduced in the CA1 and cortex of the hemisphere injured brain.Finally,the down-regulating anti-apoptotic Bcl-2 and up-regulating pro-apoptotic Bax proteins were obviously suppressed.CONCLUSION TSSN plays a potential neuroprotective role against a severe injury induced by transient focal cerebral ischemic reperfusion in a rat experimental model,and this role may be mediated by its antiinflammatory and anti-apoptotic actions.
基金National Natural Science Foundation of China(81170148)and Major Project of Natural Science Foundation of the Department of Education of Anhui Province(KJ2019ZD32)。
文摘OBJECTIVE To predict the potential targets of hyperoside(Hyp)on improving ischemia/reperfusion injury by network pharmacology,and explore its possible mechanism combined with related literature.METHODS The action targets of Hyp and ischemia/reperfusion injury were obtained by TCMSP,Swiss Target Prediction,Pharm Mapper,Similarity ensemble approach,Online Mendelian Inheritance in Man,DisGENT and database.The common targets of drugs and diseases were screened by Omishare and STRING database respectively,and the protein-protein interaction(PPI)network map was constructed.Then the interaction network between Hyp and disease targets was constructed by Cytoscape software and topological cross-linking analysis was carried out.Then the interaction network between Hyp and disease targets was constructed and cross-linked analysis was carried out by using Cytoscape software.The gene ontology(GO)of the core target was analyzed by David database,and then the related pathways of the core target were enriched by KEGG database.RESULTS A total of 54 GO enrichment processes were obtained by GO enrichment analysis of 44 common genes,including 38 biological processes(BP),15 cell composition(CC)processes,and 1 molecular functional(MF)process.43 items were obtained by signal pathway enrichment analysis in KEGG database.CONCLUSION It is suggested that the mechanism of Hyp may be related to PI3K-Akt,RAP1,RAS,VEGF and other signal transduction pathways.The above results laid a theoretical foundation for the study of the mechanism and clinical application of the treatment of ischemia/reperfusion injury.
基金Fund of Dean of Huachuang Institute of Areca Research-Hainan(HCBL2020YZ-012)。
文摘The morbidity and mortality of cardiovascular diseases are very high,which has attracted more and more attention all over the world.Common treatment methods for clinical treatment of acute myocardial infarction include direct percutaneous coronary intervention and coronary artery bypass grafting,which can quickly restore blocked coronary blood flow and reduce the infarct size.However,the inevitable ischemia/reperfusion injury will occur during the recovery of coronary blood flow,its pathological mechanism is complicated,and the Western medicine countermeasures are very limited.Among the current drugs for the treatment of cardiovascular diseases,traditional Chinese medicine has become a research hotspot due to its multiple targets,safety,and low side effects.Ginger is the fresh rhizome of Zingiber officinale Rosc.,a perennial herbaceous plant in the ginger family.It is a dual-purpose resource of medicine and food.Ginger has the functions of relieving the appearance and dispelling cold,warming up and relieving vomiting,resolving phlegm and relieving cough,and relieving fish and crab poison.The chemical components of ginger mainly include volatile oil,gingerol,diphenylheptane,etc..Among them,6-gingerol,as the main active component of gingerols,has obvious pharmacological effects in myocardial protection,anti-oxidation,anti-inflammatory,etc..Studies have shown that 6-gingerol protects myocardium mainly through anti-oxidative stress,anti-inflammatory,inhibiting cell apoptosis,and preventing calcium influx.①Anti-oxidative stress:oxidative stress is a state where oxidation and anti-oxidation in the body are out of balance,and it is also an important factor leading to myocardial damage.Many studies have confirmed that 6-gingerol has an antioxidant effect,and it is considered a natural antioxidant.6-gingerol can significantly reduce the degree of oxidative stress and the level of reactive oxygen species caused by cardiomyocyte damage,and has a significant cardioprotective effect.②Anti-inflammatory:inflammation can cause substantial cell damage and organ dysfunction,which is another important cause of myocardial damage.6-gingerol can reduce the levels of inflammatory factors such as interleukin-6,interleukin-1β,and tumor necrosis factor-αin cardiomyocytes,and at the same time inhibit the TLR4/NF-κB signaling pathway,an important regulatory pathway of inflammation,showing that it may improve myocardial damage through anti-inflammatory effects.③Inhibition of apoptosis:apoptosis is a complex and orderly process in the autonomous biochemical process of cells,and one of the main mechanisms of myocardial injury.This process can be roughly divided into three pathways:mitochondria,endoplasmic reticulum,and death receptors.Among them,the mitochondrial pathway plays an important role,and Bcl-2 and Bax located upstream of this pathway can regulate the entire process of cell apoptosis by regulating the permeability of the mitochondrial membrane.Studies have found that the preventive application of 6-gingerol can reduce cell damage,reduce the number of apoptotic cells,reduce the activity of Bax and caspase-3,and increase the expression of Bcl-2.Therefore,6-gingerol pretreatment can reduce the damage of cardiomyocytes,and its mechanism may be related to the inhibition of apoptosis.④Prevent calcium influx:calcium overload is involved in the pathogenesis of myocardial ischemic injury,which may be related to excessive contracture,arrhythmia,and mitochondrial Ca2+accumulation that impairs myocardial function.6-gingerol inhibits the increase of intracellular Ca2+concentration by inhibiting L-type calcium current,thereby reducing extracellular Ca2+influx,thereby avoiding calcium overload and playing a cardioprotective effect.In summary,6-gingerol can effectively treat and improve myocardial ischemia/reperfusion injury,and it has great development potential in the fields of medicine and health products.
基金National Natural Science Foundation of China(81173596)and Major Project of Natural Science Foundation of the Department of Education of Anhui Province(KJ2019ZD32)。
文摘OBJECTIVE To explore the effect of total flavonoids of Rhododendra simsii(TFR)on improving cerebral ischemia/reperfusion injury(CIRI)and its relationship with STIM/Orai-regulated operational Ca^(2+)influx(SOCE)pathway.METHODS Oxygen-glucose deprivation/reoxygenation(OGD/R)PC12 cells were used to simulate CIRI in vitro,and the intracellular Ca^(2+)concentration and apoptosis rate of PC12 cells were detected by laser confocal microscope and flow cytometry,respectively.The regulation of STIM/Orai on SOCE was analyzed by STIM/Orai gene silencing and STIM/O rai gene overexpression.The CIRI model was established by MCAO in SD rats.The activities of inflammatory cytokines IL^(-1),IL-6 and TNF-αin serum were detected by ELISA.The pathological changes of ischemic brain tissue and the infarction of rat brain tissue were detected by HE staining and TTC staining.The protein and mRNA expression levels of STIM1,STIM2,Orai1,caspase-3 and PKB in brain tissue were detected by Western blotting and RT-qPCR,respectively.RESULTS The results of in vitro experiment showed that the fluorescence intensity of Ca^(2+)and apoptosis rate in PC12 cells treated with TFR were significantly lower than those in OGD/R group,and this trend was enhanced by SOCE antagonist 2-APB.STIM1/STIM2/Orai1 gene silencing significantly reduced apoptosis and Ca^(2+)overload in OGD/R model,while TFR combined with overexpression of STIM1/STIM2/Orai1 aggravated apoptosis and Ca2+overload.In the in vivo experiment,TFR significantly reduced the brain histopathological damage,infarction of brain tissue,the contents of IL^(-1),IL-6 and TNF-αin the serum in MCAO rats and down-regulated the expression of STIM1,STIM2,Orai1 and caspase-3 protein and mRNA in the brain tissue,and up-regulated the expression of PKB.The above effects were enhanced by the addition of 2-APB.CONCLUSION The above results indicate that TFR may reduce the contents of inflammatory factors and apoptosis,decrease Ca2+overload and ameliorate brain injury by inhibiting SOCE pathway mediated by STIM and Orai,suggesting that it has a protective effect against subacute CIRI.
文摘Aim Salvia miltiorrhiza Bunge （SM） and lignum dalbergiae odoriferae （DO） are both traditional Chi- nese medicine that have cardioprotective effects. Here, we further examined the combined effects of SM and DO on rat myocardial ischemia/reperfusion injury. The possible mechanism of SM and DO also were elucidated. Methods DO was divided into aqueous extract of lignum dalbergiae odoriferae （DOW） and lignum dalbergiae odoriferae oil （DOO）. Sprague-Dawley rats were randomized to seven groups： sham group, model group, treatment groups inclu- ding SM （10 g · kg^-1）, DOW （5 g · kg^-1）, DOO （0.5 ml · kg^-1）, SM ＋ DOW （10 g · kg^-1 ＋ 5 g · kg^-1）, SM ＋ DOO （ 10 g · kg^-1 ＋ 0. 5 ml · kg^-1）. Rats were pretreated with homologous drug for 7 days and then subjec- ted to 30 rain of ischemia followed by 180 rain of reperfusion. Electrocardiogram （ECG） and heart rate were moni- tored and recorded continuously. At the end of reperfusion, blood samples were collected to determine the serum levels of creatine kinase-MB （CK-MB） and lactate dehydrogenase （LDH）. Hearts were harvested to assess heart- body rate, infarct size and histopathological changes as well. Maximum and minimum effective points were deter- mined by measuring indicators associate with myocardial injury at different time-points of reperfusion （Smin, 15min, 30min, 45rain, 60min, 120min, 180min）. The potential therapeutic mechanism of SM and SM ＋ DOO were carried out by detecting superoxide dismutase （SOD）, malondialdehyde （MDA）, tumor necrosis factor-alpha （TNF-alpha） and interleukin 6 （IL-6）. Results The results showed SM and DO can ameliorate cardiac function respectively, and this cardioprotective effect was further strengthened by their combinations. Among all the combi- nations, SM ＋ DOO showed predominant potential to improve ECG and heart rate, reduce heart-body rate （28.5% ＋ 1.4% , P 〈 0.01 vs model） and myocardial infarct size （ 20.96% ＋ 1.61% , P 〈 0.01 vs model, P 〈 0.05 vs SM） , attenuate histopathological damage, decrease the levels of CK-MB and LDH （P 〈 0.01 vs model, P 〈 0.05 vs SM）. The maximum effective points of SM and SM ＋ DOO were 15min and 30rain respectively, and the minimum effective points of them were 180rain. In reducing serum level of MDA, TNF-alpha, IL-6 and increasing SOD activ- ity, SM ＋ DOO was similar to SM. Conclusion The results of this study indicated that SM ＋ DOO have combined effects that are highly effective than single pretreatment against myocardial ischemie reperfusion injury in rats. The possible mechanism of SM and DO were likely through its anti-oxidant and anti-inflammatory properties, and thus may be an effective and promising medicine for both prophylaxis and treatment of ischemic heart disease.
基金National Natural Science Foundation of China(81173596)Natural Science Foundation of the Department of Education of Anhui Province(KJ2015A157)
文摘OBJECTIVE To investigate regulatory effects of hyperoside(Hyp)on IP3/PKC/TRPV4 pathway in rat cerebral basilar artery(CBA)subjected to global cerebral ischemia-reperfusion(I/R).METHODS The model of global cerebral I/R in rats was established by four-vessel occlusions methods.The treated rats were administrated with Hyp(50 mg·kg^-1)group,Hyp(50 mg·kg^-1)+HC-067047(10 mg·kg^-1),Hyp(50 mg·kg^-1)+2APB(2 mg·kg^-1),Hyp(50 mg·kg^-1)+BisI(2.5 mg·kg^-1),Hyp(50 mg·kg^-1)+2APB(2 mg·kg^-1)+BisI(2.5 mg·kg^-1).Hematoxylin-eosin(HE)and Nissl staining were performed and the contents of methane dicarboxylic aldehyde(MDA),neuron-specific enolase(NSE),S100β and the activity of lactic dehydrogenase(LDH)in serum were measured by enzyme-linked immunosorbnent assay(ELISA).The specific blocker N-nitro-L-arginine-methyl-ester(L-NAME)and indomethacin(Indo)were used to delete the prostacyclin(PGI2)and nitric oxide(NO)dependent relaxation.The protein expression level of TRPV4 was detected by Western blotting.Ca2+intensity in vascular smooth muscle cells was measured by confocal laser scanning microscope and flow cytometry was performed to observe the apoptosis of CBA endothelial cells after in vivo administration.RESULTS Hyp induced a dose-dependent relaxation of CBA in IR rats via a PGI2 and NO independent manner,as evidenced by alleviated patho⁃logical changes and up-regulated expression of TRPV4 protein in the endothelial cells from cerebral vessels.Hyp signifi⁃cantly reduced the contents of MDA,NSE,S100βand the activity of LDH in serum and decreased the fluorescence intensity of Ca2+in cerebral vascular smooth muscle cells by in vivo administration.The apoptotic rate of endothelial cells in Hyp treated group was significantly less than that in IR group.CONCLUSION Hyp does in fact ameliorate I/R injury by regulatingIP3/PKC/TRPV4 pathway.
基金The project supported by National Key Basic Research Developmentplans(2012CB723503)
文摘OBJECTIVE To investigate the prevention and its mechanism of Naojian Tablet,the adjusted prescription from Buyang Huanwu Decoction,during focal ischemic brain injury condition,and study on the main bioactive substance for this function.METHODS To apply the modified middle cerebral artery occlusion(MCAO)model in rats,immunohistochemistry,RT-PCR and Western blotting were applied to determine the expression of CDK5,CDK4/cyclinD1 in the hippocampus tissues in MCAO rat by treating with Naojian Tablet.On the other hand,the processes of metabolism and disposition of several active components in Naojian Tablet and Buyang Huanwu Decoction were analyzed and characterized by using HPLC-Q-TOF method.RESULTS Naojian Tablet(ig,2.41g·kg-1)could significantly decrease the levels of CDK5,CDK4/cyclinD1 in hippocampus tissues in MACO rat after treated by 3d,7d,14 d,28d(P<0.05).The similar mechanism was observed in the Buyang Huanwu Decoction treated MACO rat.Calycosin,fermononetin,senkyunolide and ligustilide in Naojian Tablet was determined simultaneously as the main active ingredient of Naojian Tablet.CONCLUSION The above-mentioned in vivo studies suggested that Naojian Tablet maybe play its prevention effect on nerve cells via its acting on CDK5 signaling and cell cycle pathway,which verified the multi-target and multi active pathways efficiency of tradition Chinese medicine in treating diverse disease.
文摘Myocardial infarction resulting from coronary atherosclerosis is the leading cause of death in modern soci- ety. Reperfusion is an essential treatment to salvage ischemia myocardium from necrosis, while it also leads to addi- tional damage. Therefore, exploring effective medicines to protect the heart from post-ischemic injury is one of the major objectives of cardiovascular research. Berbamine is a nature compound of bisbenzylisochinoline alkaloids from Barberry. We found that it displays positive inotropic and lusitropic effects at lower concentrations by increasing myofilament Ca2＋ sensitivity via a PKCe-dependent signaling pathway. Moreover, berbamine preconditioning con- fers cardioprotection against ischemia/reperfusion （I/R） injury by attenuating the Ca2＋ overloading and preventing the calpain activation through the activating of PI3K-Akt-GSK3β pathway and subsequently opening of the mitoKATP channel. Furthermore, we demonstrate that berbamine postconditioning conferred the cardioprotective effect against I/R injury by the regulation of autophagy. These findings reveal new roles and mechanisms of berbamine in the heart and cardioprotection against I/R injury.
文摘Aim Reduction of Sheng-Nao-Kang decoction （RSNK）, is a modified traditional Chinese medicinal formula of Sheng-Nao-Kang pill preparation, which is protective in rats against focal cerebral ischemia/reperfusion （I/R） injury. In the current study, we investigate the protective effect of RSNK against apoptosis and oxidative damage induced by cerebral I/R and explore the underlying mechanisms. Cerebral I/R injury was induced by in- traluminal middle cerebral artery occlusion （MCAO） for 2 h followed by reperfusion for 24 h in adult male Sprague- Dawley rats. Rats were randomized into seven groups （n- 8）： Sham group, I/R group, RSNK-treated groups （ 0.7 g · kg ^- 1, 1 . 4 g · kg ^- 1 and 2. 8 g · kg^ - 1 ） , nimodipine （NMP） -treated group and Whitmania pigra Whitman （WW）-treated group. Neurological deficit scores, cerebral humidity content and cerebral infarction volume were measured after the 24 h reperfusion. Malondialdehyde （ MDA）, superoxide dismutase （ SOD）, catalase （ CAT）, inducible nitric oxide synthase （iNOS） and total nitric oxide synthase （TNOS） in serum were measured by assay kits for biochemical analysis. Histological structures of the cortex of the ipsilateral ischemic cerebral hemisphere in rats were observed by Nissl staining. The caspase-3 protein content in the hippocampus and cortex was detected by immunohistochemistry. Additionally, Bax and Bcl-2 protein expressions in the injured brain were evaluated by Western blot. RSNK administration not only markedly improved neurological deficit scores, but also reduced cere- bral humidity content and cerebral infarction volume, lowered MDA content, up-regulated SOD and CAT levels, down-regulated iNOS and TNOS levels, restrained the expression of caspase-3 positive protein and alleviated the Bax and Bcl-2 protein expressions.
