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
OBJECTIVE To investigates the effects of imperatorin on the oxidative stress in the cerebral cortex and hippocampus after focal cerebral ischemia/reperfusion injury.METHODS Transient focal cerebral ischemia/reperfusio...OBJECTIVE To investigates the effects of imperatorin on the oxidative stress in the cerebral cortex and hippocampus after focal cerebral ischemia/reperfusion injury.METHODS Transient focal cerebral ischemia/reperfusion model in male Sprague-Dawley rats was induced by 2 h middle cerebral artery occlusion followed by 24 h reperfusion.Imperatorin(1.25 and 2.5 mg·kg-1)or vehicle were administered intraperitoneally at 1,5 and 9 h after the onset of ischemia.At 24 h after reperfusion,the biomarkers of oxidative stress such as the levels of reactive oxygen species(ROS),lipid peroxidation products malondialdehyde(MDA),nitric oxide(NO)and total antioxidant capacity(T-AOC),the activities of inducible nitric oxide synthase(iN OS),superoxide dismutase(SOD)and catalase(CAT)in the cerebral cortex and hippocampus were observed.We also assessed the nuclear factor erythroid 2-related factor 2(Nrf2),heme oxygenase-1(HO-1),and the NAD(P)H-quinone oxidoreductase 1(NQO-1)protein expression by Western blot.RESULTS As compared to vehicle-treated animals,imperatorin treatment significantly reduced the ROS,MDA,NO levels and i NOS activity,increased T-AOC and the activities of SOD and CAT.Furthermore,imperatorin treatment also significantly induced the nuclear translocation of Nrf2,enhanced the protein expression of HO-1 and NQO-1 in the cerebral cortex and hippocampus.CONCLUSION Our findings indicate that imperatorin can protect the brain against the excessive oxidative stress induced by cerebral ischemia/reperfusion through activation of Nrf2 signaling pathway.展开更多
OBJCETIVE Epidemiologic studies have demonstrated that consumption of moderate amounts of red wine is associated with significant reductions in incidences of cardiovascular and cerebrovascular diseases,which may be re...OBJCETIVE Epidemiologic studies have demonstrated that consumption of moderate amounts of red wine is associated with significant reductions in incidences of cardiovascular and cerebrovascular diseases,which may be related to alcohol in red wine.Our previous study demonstrated that ethanol ingestion 24 h prior to induction of cerebral ischemic/reperfusion(I/R)reduced delayed neuronal death(DND).Our most recent results supported a role for big Ca2+-sensitive K+channel(BKCa channel)activation in the neuroprotective effects of ethanol preconditioning(Et OH-PC)in global cerebral I/R.Therefore,we hypothesis that moderate Et OH-PC activates BKCa channel to protect brain damage induced by focal cerebral I/R.This project will utilize focal cerebral I/R animal model to explore the function of BKCa channel in Et OH-PC protection in vivo levels by means of pharmacological intervention such as BKCa channel opene(rNS11021,NS)and blocke(rpaxilline,PX).The results will provide theoretical evidence for neuroprotective effect of moderate alcohol preconditioning against ischemic stroke,and the conclusion will also bring to a concept that extrinsic moderate ethanol preconditioning may activate intrinsic protective mechanism in the brain.METHODS The SD rat were randomly divided into the following six groups(n=10):sham,I/R,Et OH-PC+I/R,NS11021-PC+I/R,paxilline+Et OH-PC+I/R,Paxilline+NS11021-PC+I/R.Both Et OH-PC and NS11021-PC(0.1mg·kg-1;ip)were induced 24 h before I/R.The volume of 95%ethanol to be instilled(inμL)was calculated as follows:〔body weight(g)×0.6〕+0.3.This volume of ethanol was mixed in 0.3 m L of sterile distilled water just before administration to the animals by gavage.The Paxilline(2.5 mg·kg-1;ip)was administered 10min beforeEt OH-PC and NS11021-PC.The right middle cerebral artery occlusion(MCAO)was produced by inversion of a 4-0-nylon filament.The filament was withdrawn 2 h after onset of MCAO and then reperfused.Neurological deficits and infarct volume were measured 24 h after I/R.Another 36 rats were randomly divided into 6 groups as above,6 in each group.DWI were performed 2h after ischemic and T2WI MRI were performed 24 h after I/R to observe the infarct volume of brain and the penumbra volume of brain in each group.Then rats were killed and detected the apoptotic cell death and degeneration of neurons.RESULTS Compared to I/R group,the neurological score(P<0.01),the infarct volume of brain(P<0.01),the infarct volume of ischemic penumbra(P<0.01),the percentage of apoptotic cell death(P<0.01)and the percentage of degenerative neurons(P<0.01)were significantly decreased after ethanol preconditioning,while these changes were reversed by paxilline(P<0.05);compared to I/R group,the neurological score(P<0.01),the infarct volume of brain(P<0.01),the infarct volume of ischemic penumbra(P<0.01),the percentage of apoptotic cell death(P<0.01)and the percentage of degenerative neurons(P<0.01)were significantly decreased after NS11021 preconditioning,while these changes were reversed by paxilline(P<0.05).CONCLUSION Our results show that moderate alcohol preconditioning activates BKCa channels to protect brain damage induced by focal cerebral I/R.展开更多
We recently reported that nucleotide-binding oligomerization domain （NOD） 2, an important cytoplasmic pattern recognition receptor, is involved in cerebral ischemia-reperfusion （I/R） injury. β-arrestins, in addit...We recently reported that nucleotide-binding oligomerization domain （NOD） 2, an important cytoplasmic pattern recognition receptor, is involved in cerebral ischemia-reperfusion （I/R） injury. β-arrestins, in addition to regulate desensitization of G protein-coupled receptors （GPCRs） , have emerged as potential mediators of innate im- mune activation. However, the role and mechanism of β-arrestin2 in NOD2-triggered signaling in the cerebral I/R remain to be established. Methods BV2 cells were transfected with either β-arrestin2-shRNA plasmid or β-arres- tin2 full-length plasmid and control vector. Middle cerebral artery occlusion （MCAO） was induced in male wild- type mice and in wild type （WT） and β-arrestin2 deficient mice. Results muramyl dipeptide （MDP）, an extrin- sic ligand of NOD2, significantly increased the expression of TRAF6 and COX-2 and enhanced the activation of NF- KB in the microglia time-dependently. MDP stimulation also promoted the expression and activation of MMP-9 time- dependently, but did not affect MMP-2 obviously. Additionally, β-arrestin 2 interacted with TRAF6 after MDP stim- ulation rapidly. Overexpression of β-arrestin2 inhibited NF-KB and MMP-9 activation and COX-2 upregulation in- duced by MDP, while silence of β-arrestin2 enhanced NOD2-triggered inflammatory signaling. Finally, Deletion of β-arrestin 2 markedly aggravated brain infarction, neurological deficit and inflammation induced by MDP in mice subjected to MCAO. Conclusion The results provide the first evidence that β-arrestin 2 is an essential negatively regulator of NOD2 triggered inflammatory signaling in the cerebral I/R injury.展开更多
Objective:To investigate the influence of electroacupuncture(EA) combined with repetitive transcranial magnetic stimulation(rTMS) on the temporal profile of nestin expression after induction of focal cerebral ischemia...Objective:To investigate the influence of electroacupuncture(EA) combined with repetitive transcranial magnetic stimulation(rTMS) on the temporal profile of nestin expression after induction of focal cerebral ischemia in adult rats and to explore the mechanism of EA combined with rTMS in treating ischemic brain injury.Method:The model of transient focal ischemia was produced by occlusion of middle cerebral artery.Seventy-five Wistar rats were randomly divided into normal group,model group,EA group,rTMS group and EA +rTMS group.The neurologic impairment rating and ability of learning and memory were observed at the 7th、14th and 28th d after infarction respectively.Meanwhile,Western blotting was used to observe the number of nestin expression positive cells.Result:Nestin-positive cells were found in cortex,subgranular zone(SGZ),subventricular zone(SVZ) of the ipsilateral side at different time points after cerebral ischemia.The number of nestin-positive cells peaked at the 7th d,began to decrease at the 14th d and was significantly higher in EA+rTMS group than that in model group(P< 0.05),then almost reached normal at the 28th d.The improvement of neural motor function deficits as well as the indexes of learning and memory were more obvious in EA+rTMS group compared with model group(P<0.01,P<0.05).These effects were most obvious in EA +rTMS group compared with the EA and rTMS group(P<0.05).Conclusion:EA and rTMS possess the potency of building up and can increase the number of nestin-positive cells in some brain regions after focal cerebral ischemia,which might be one of the important mechanisms of EA combined with rTMS in treating ischemia brain injury.展开更多
Aim Shengmai injection （SMI） , a Chinese patent medicine deprived from an ancient Chinese herbal compound Shengmai san, which is used extensively for the treatment of cardiovascular and cerebrovascular disease in cl...Aim Shengmai injection （SMI） , a Chinese patent medicine deprived from an ancient Chinese herbal compound Shengmai san, which is used extensively for the treatment of cardiovascular and cerebrovascular disease in clinic. To determine the neuroprotective effect of SMI, the effect and the relevant mechanism of SMI had been inves- tigated on cerebral ischemia-reperfusion injury in mice. Methods Right middle cerebral artery was occluded by in- serting a thread through intemal carotid artery for 1 h, and then reperfusion for 24 h in mice. Neuroprotective effect was testified using transmission electron microscopic examination, evaluation of infarct volume and neurological defi- cits. Related mechanism was evaluated by western blotting. The SMI was injected intraperitoneally after ischemia for 1 h at doses of 1.42, 2. 84 and 5.68 g · kg^-1. The control group received saline as vehicle of SMI. Results SMI （ 1.42, 2. 84 and 5. 68 g · kg^-1） could significantly reduced the infarct volume, SMI （5.68 g · kg^-1） could signifi- cantly improved the neurological deficits, as well as the neuron＇s morphology change. SMI （5.68 g · kg^-1） could significantly inhibited the autophagy-related proteins： Beclinl and LC3. SMI （5. 68 g · kg^-1） remarkably inhibited the phosphorylation of adenosine monophosphate activated protein kinase （AMPK）, and down-regulated the phospho- rylation of mammalian target of rapamycin （roTOR） and Jun N-terminal kinase （JNK） after 24 h reperfusion. Con-clusion The results indicated that SMI elicits potent protection against cerebral ischemia/reperfusion injury, which may partly be due to the inhibition in autophagy and related signal pathways.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
OBJECTIVE To investigate the effects of IMM-H004 on permanent focal cerebral ischemia injury and associated cardiopulmonary complications,further elucidating the molecular mechanisms.METHODS The effects of IMM-H004 we...OBJECTIVE To investigate the effects of IMM-H004 on permanent focal cerebral ischemia injury and associated cardiopulmonary complications,further elucidating the molecular mechanisms.METHODS The effects of IMM-H004 were investigated in wild-type(WT) and CKLF1-/-rats.The effects of IMM-H004 on ischemic stroke injury and its cardiopulmonary complications were determined using 2,3,5-triphenyltetrazolium chloride(TTC) staining,behavior tests,magnetic resonance imaging(MRI)scans,enzyme-linked immunosorbent assay(ELISA),Nissl staining,and histo-pathological examination.Multiple molecular experiments including immunohistological staining,immunofluorescence staining,quantitative RT-PCR,Western blotting,and co-immunoprecipitation assays were used to elucidate the underlying mechanisms.RESULTS IMM-H004 treatment provided significant protection against ischemic stroke-induced brain injury and associated cardiopulmonary complications,through CKLF1-depedent-anti-inflammation pathway in rats.IMM-H004 downregulated the amount of CKLF1 and disturbed the combination between CKLF1 and C-C chemokine receptor type 4,suppressing the inflammatory response and protecting the damaged organs in ischemic setting.CONCLUSION This preclinical study established efficacy of IMM-H004 as a potential therapeutic medicine for ischemic stroke and associated cardiopulmonary complications.The protective effects of IMM-H004 may due to its specific mechanism through CKLF1.These results support further efforts to develop IMM-H004 for human clinical trials in acute cerebral ischemia,especially for patients who are not suitable for reperfusion therapy.展开更多
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.
基金supported by National Natural Science Foundation of China(81060269 and81360492)Natural Science Foundation of Jiangxi Province of China(20122BAB205036)
文摘OBJECTIVE To investigates the effects of imperatorin on the oxidative stress in the cerebral cortex and hippocampus after focal cerebral ischemia/reperfusion injury.METHODS Transient focal cerebral ischemia/reperfusion model in male Sprague-Dawley rats was induced by 2 h middle cerebral artery occlusion followed by 24 h reperfusion.Imperatorin(1.25 and 2.5 mg·kg-1)or vehicle were administered intraperitoneally at 1,5 and 9 h after the onset of ischemia.At 24 h after reperfusion,the biomarkers of oxidative stress such as the levels of reactive oxygen species(ROS),lipid peroxidation products malondialdehyde(MDA),nitric oxide(NO)and total antioxidant capacity(T-AOC),the activities of inducible nitric oxide synthase(iN OS),superoxide dismutase(SOD)and catalase(CAT)in the cerebral cortex and hippocampus were observed.We also assessed the nuclear factor erythroid 2-related factor 2(Nrf2),heme oxygenase-1(HO-1),and the NAD(P)H-quinone oxidoreductase 1(NQO-1)protein expression by Western blot.RESULTS As compared to vehicle-treated animals,imperatorin treatment significantly reduced the ROS,MDA,NO levels and i NOS activity,increased T-AOC and the activities of SOD and CAT.Furthermore,imperatorin treatment also significantly induced the nuclear translocation of Nrf2,enhanced the protein expression of HO-1 and NQO-1 in the cerebral cortex and hippocampus.CONCLUSION Our findings indicate that imperatorin can protect the brain against the excessive oxidative stress induced by cerebral ischemia/reperfusion through activation of Nrf2 signaling pathway.
基金The project supported by NSFC(81171079,81271312)
文摘OBJCETIVE Epidemiologic studies have demonstrated that consumption of moderate amounts of red wine is associated with significant reductions in incidences of cardiovascular and cerebrovascular diseases,which may be related to alcohol in red wine.Our previous study demonstrated that ethanol ingestion 24 h prior to induction of cerebral ischemic/reperfusion(I/R)reduced delayed neuronal death(DND).Our most recent results supported a role for big Ca2+-sensitive K+channel(BKCa channel)activation in the neuroprotective effects of ethanol preconditioning(Et OH-PC)in global cerebral I/R.Therefore,we hypothesis that moderate Et OH-PC activates BKCa channel to protect brain damage induced by focal cerebral I/R.This project will utilize focal cerebral I/R animal model to explore the function of BKCa channel in Et OH-PC protection in vivo levels by means of pharmacological intervention such as BKCa channel opene(rNS11021,NS)and blocke(rpaxilline,PX).The results will provide theoretical evidence for neuroprotective effect of moderate alcohol preconditioning against ischemic stroke,and the conclusion will also bring to a concept that extrinsic moderate ethanol preconditioning may activate intrinsic protective mechanism in the brain.METHODS The SD rat were randomly divided into the following six groups(n=10):sham,I/R,Et OH-PC+I/R,NS11021-PC+I/R,paxilline+Et OH-PC+I/R,Paxilline+NS11021-PC+I/R.Both Et OH-PC and NS11021-PC(0.1mg·kg-1;ip)were induced 24 h before I/R.The volume of 95%ethanol to be instilled(inμL)was calculated as follows:〔body weight(g)×0.6〕+0.3.This volume of ethanol was mixed in 0.3 m L of sterile distilled water just before administration to the animals by gavage.The Paxilline(2.5 mg·kg-1;ip)was administered 10min beforeEt OH-PC and NS11021-PC.The right middle cerebral artery occlusion(MCAO)was produced by inversion of a 4-0-nylon filament.The filament was withdrawn 2 h after onset of MCAO and then reperfused.Neurological deficits and infarct volume were measured 24 h after I/R.Another 36 rats were randomly divided into 6 groups as above,6 in each group.DWI were performed 2h after ischemic and T2WI MRI were performed 24 h after I/R to observe the infarct volume of brain and the penumbra volume of brain in each group.Then rats were killed and detected the apoptotic cell death and degeneration of neurons.RESULTS Compared to I/R group,the neurological score(P<0.01),the infarct volume of brain(P<0.01),the infarct volume of ischemic penumbra(P<0.01),the percentage of apoptotic cell death(P<0.01)and the percentage of degenerative neurons(P<0.01)were significantly decreased after ethanol preconditioning,while these changes were reversed by paxilline(P<0.05);compared to I/R group,the neurological score(P<0.01),the infarct volume of brain(P<0.01),the infarct volume of ischemic penumbra(P<0.01),the percentage of apoptotic cell death(P<0.01)and the percentage of degenerative neurons(P<0.01)were significantly decreased after NS11021 preconditioning,while these changes were reversed by paxilline(P<0.05).CONCLUSION Our results show that moderate alcohol preconditioning activates BKCa channels to protect brain damage induced by focal cerebral I/R.
文摘We recently reported that nucleotide-binding oligomerization domain （NOD） 2, an important cytoplasmic pattern recognition receptor, is involved in cerebral ischemia-reperfusion （I/R） injury. β-arrestins, in addition to regulate desensitization of G protein-coupled receptors （GPCRs） , have emerged as potential mediators of innate im- mune activation. However, the role and mechanism of β-arrestin2 in NOD2-triggered signaling in the cerebral I/R remain to be established. Methods BV2 cells were transfected with either β-arrestin2-shRNA plasmid or β-arres- tin2 full-length plasmid and control vector. Middle cerebral artery occlusion （MCAO） was induced in male wild- type mice and in wild type （WT） and β-arrestin2 deficient mice. Results muramyl dipeptide （MDP）, an extrin- sic ligand of NOD2, significantly increased the expression of TRAF6 and COX-2 and enhanced the activation of NF- KB in the microglia time-dependently. MDP stimulation also promoted the expression and activation of MMP-9 time- dependently, but did not affect MMP-2 obviously. Additionally, β-arrestin 2 interacted with TRAF6 after MDP stim- ulation rapidly. Overexpression of β-arrestin2 inhibited NF-KB and MMP-9 activation and COX-2 upregulation in- duced by MDP, while silence of β-arrestin2 enhanced NOD2-triggered inflammatory signaling. Finally, Deletion of β-arrestin 2 markedly aggravated brain infarction, neurological deficit and inflammation induced by MDP in mice subjected to MCAO. Conclusion The results provide the first evidence that β-arrestin 2 is an essential negatively regulator of NOD2 triggered inflammatory signaling in the cerebral I/R injury.
基金Supported by the National Nature Science Foundation of China(No.30672216)Project of Wuhan Hygiene Bureau(No.WX08A01,No.WZ08B02)
文摘Objective:To investigate the influence of electroacupuncture(EA) combined with repetitive transcranial magnetic stimulation(rTMS) on the temporal profile of nestin expression after induction of focal cerebral ischemia in adult rats and to explore the mechanism of EA combined with rTMS in treating ischemic brain injury.Method:The model of transient focal ischemia was produced by occlusion of middle cerebral artery.Seventy-five Wistar rats were randomly divided into normal group,model group,EA group,rTMS group and EA +rTMS group.The neurologic impairment rating and ability of learning and memory were observed at the 7th、14th and 28th d after infarction respectively.Meanwhile,Western blotting was used to observe the number of nestin expression positive cells.Result:Nestin-positive cells were found in cortex,subgranular zone(SGZ),subventricular zone(SVZ) of the ipsilateral side at different time points after cerebral ischemia.The number of nestin-positive cells peaked at the 7th d,began to decrease at the 14th d and was significantly higher in EA+rTMS group than that in model group(P< 0.05),then almost reached normal at the 28th d.The improvement of neural motor function deficits as well as the indexes of learning and memory were more obvious in EA+rTMS group compared with model group(P<0.01,P<0.05).These effects were most obvious in EA +rTMS group compared with the EA and rTMS group(P<0.05).Conclusion:EA and rTMS possess the potency of building up and can increase the number of nestin-positive cells in some brain regions after focal cerebral ischemia,which might be one of the important mechanisms of EA combined with rTMS in treating ischemia brain injury.
文摘Aim Shengmai injection （SMI） , a Chinese patent medicine deprived from an ancient Chinese herbal compound Shengmai san, which is used extensively for the treatment of cardiovascular and cerebrovascular disease in clinic. To determine the neuroprotective effect of SMI, the effect and the relevant mechanism of SMI had been inves- tigated on cerebral ischemia-reperfusion injury in mice. Methods Right middle cerebral artery was occluded by in- serting a thread through intemal carotid artery for 1 h, and then reperfusion for 24 h in mice. Neuroprotective effect was testified using transmission electron microscopic examination, evaluation of infarct volume and neurological defi- cits. Related mechanism was evaluated by western blotting. The SMI was injected intraperitoneally after ischemia for 1 h at doses of 1.42, 2. 84 and 5.68 g · kg^-1. The control group received saline as vehicle of SMI. Results SMI （ 1.42, 2. 84 and 5. 68 g · kg^-1） could significantly reduced the infarct volume, SMI （5.68 g · kg^-1） could signifi- cantly improved the neurological deficits, as well as the neuron＇s morphology change. SMI （5.68 g · kg^-1） could significantly inhibited the autophagy-related proteins： Beclinl and LC3. SMI （5. 68 g · kg^-1） remarkably inhibited the phosphorylation of adenosine monophosphate activated protein kinase （AMPK）, and down-regulated the phospho- rylation of mammalian target of rapamycin （roTOR） and Jun N-terminal kinase （JNK） after 24 h reperfusion. Con-clusion The results indicated that SMI elicits potent protection against cerebral ischemia/reperfusion injury, which may partly be due to the inhibition in autophagy and related signal pathways.
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.
基金The project supported by ZYBZH-Y-HUN-24National Natural Science Foundation of China(81730096+6 种基金U1402221)National Mega-project for Innovative Drugs(2018ZX09711001-002-0072018ZX09711001-003-0052018ZX09711001-009-013)CAMS Innovation Fund for Medical Sciences(2016-I2M-1-004)Beijing Key Laboratory of New Drug Mechanisms and Pharmacological Evaluation Study(BZ0150)PUMC Graduate Educationand Teaching Reform Project(10023201600801)
文摘OBJECTIVE To investigate the effects of IMM-H004 on permanent focal cerebral ischemia injury and associated cardiopulmonary complications,further elucidating the molecular mechanisms.METHODS The effects of IMM-H004 were investigated in wild-type(WT) and CKLF1-/-rats.The effects of IMM-H004 on ischemic stroke injury and its cardiopulmonary complications were determined using 2,3,5-triphenyltetrazolium chloride(TTC) staining,behavior tests,magnetic resonance imaging(MRI)scans,enzyme-linked immunosorbent assay(ELISA),Nissl staining,and histo-pathological examination.Multiple molecular experiments including immunohistological staining,immunofluorescence staining,quantitative RT-PCR,Western blotting,and co-immunoprecipitation assays were used to elucidate the underlying mechanisms.RESULTS IMM-H004 treatment provided significant protection against ischemic stroke-induced brain injury and associated cardiopulmonary complications,through CKLF1-depedent-anti-inflammation pathway in rats.IMM-H004 downregulated the amount of CKLF1 and disturbed the combination between CKLF1 and C-C chemokine receptor type 4,suppressing the inflammatory response and protecting the damaged organs in ischemic setting.CONCLUSION This preclinical study established efficacy of IMM-H004 as a potential therapeutic medicine for ischemic stroke and associated cardiopulmonary complications.The protective effects of IMM-H004 may due to its specific mechanism through CKLF1.These results support further efforts to develop IMM-H004 for human clinical trials in acute cerebral ischemia,especially for patients who are not suitable for reperfusion therapy.
