OBJECTIVE To study the role of Ginkgo biloba extract-761(EGb-761)in the recovery of gait abnormality and its neuroprotective effect against the brain injury induced by permanent middle cerebral artery occlu-sion in ra...OBJECTIVE To study the role of Ginkgo biloba extract-761(EGb-761)in the recovery of gait abnormality and its neuroprotective effect against the brain injury induced by permanent middle cerebral artery occlu-sion in rats.METHODS Male Sprague Dawley rats(n=200,240-305 g)were anesthetized with 0.2%pentobarbital sodium diluted in physiological saline(2.0 m L·kg-1,ip).Then a monofilament coated with poly-L-lysine,was used to occlude the origin of the middle cerebral artery.It was inserted into the internal carotid artery lumen until it met mild resistance,approximately 20mm beyond the common carotid artery bifurcation.The suture was secured with a ligature and maintained in place until sacrifice.The same surgical procedure was conducted in sham-operated rats in which the middle cerebral artery was not occluded.Motor and behavioral changes were assessed after surgery using a five point scale.The rats securing the point scale above 2 were included in the study.The rats were randomly divided into control,and treated groups:EGb-761(20,50,and 100 mg·kg-1).The treated groups were oral y administered(10 mL·kg-1)for 28 d.On 7th,14th,21st,and 28th day the neurological scores,rotar rod test and gait assessment(the automated computer-assisted method)were performed.The brains were collected for TTC staining and histopathological analysis.RESULTS 1)Weight:On 28th day,EGb-761(20 mg·kg-1,)significantly increased the weight of the rat by^8%as compared to control(~300 g).However,at 50 mg·kg-1,and 100 mg·kg-1,a significant increase of^7-7.6%(control:~232 g),and^7.3-7%,respectively from 14 to 28 days was noted.2)Neurological scores:On 28thday,EGb-761(20,50,and 100 mg·kg-1)significantly decreased the neurological scores by^18%,~22%,~21%,respectively as compared to control(~2.07).3)Rotar rod test:On 28thday,EGb-761(50,and100 mg·kg-1)significantly increased by^69.1%,~74.1%,respectively as compared to control(~28.2).4)Gait assessment:On 7th,14th,21st,and 28thday,EGb-761(20,50,and 100 m·kg-1)significantly reduced the average body angle,on 7th,14th,21st,and 28thday,EGb-761(100 mg·kg-1)significantly increased the walk speed and reduced the average walking cycle,EGb-761(50,and 100 mg·kg-1)significantly the area of the left brain/right brain area percentage and reduced tissue pathologic neuron injury.CONCLUSION Ginkgo biloba extract EGb-761 has obvious improve behavior disorders,and has a protective neuroprotective effect against the brain injury induced by permanent middle cerebral artery occlusion.展开更多
Objective:To investigate the effects of electro-acupuncture preconditioning on focal cerebral blood flow(CBF)in artery and vein of rats with middle cerebral artery occlusion(MCAO),and to provide experimental evidences...Objective:To investigate the effects of electro-acupuncture preconditioning on focal cerebral blood flow(CBF)in artery and vein of rats with middle cerebral artery occlusion(MCAO),and to provide experimental evidences for primary prevention of ischemic stroke.Method:Eighteen male Sprague-Dawley rats(two months old)were divided into 3 groups:electro-acupuncture preconditioning group(EA group),ischemia group(Is group)and Dazhui and Baihui sham group(six rats per group).The rats in the EA group were given electro-acupuncture preconditioning at Dazhui and Baihui with 2/15 density wave current for 30 minutes per day,5 days consecutively.After preconditioning,enhanced laser speckle contrast analysis was implemented to get false color images before making middle cerebral ischemia occlusion model.Then getting false color images two hours during ischemia by laser speckle contrast analysis again.Result:④Relative CBF in focal cortical artery:There were significant differences at every time point in EA group and Is group(P<0.01,P<0.05)comparing with that before occlusion respectively.Compared with that before ischemia,there was significant difference at 120 min after ischemia in sham group(P<0.05);compared with that at 30 min after occlusion,there was significant difference at every time point in EA group from the time point 60 min after occlusion;there was significant difference at every time point in EA group and Is group(P<0.05)comparing with sham group;Compared with sham group,there was significant difference at 30min and 60 min after ischemia in EA group(P<0.05).④Relative CBF in focal cortical vein:There was significant difference at every time point in sham group comparing with that before occlusion(P<0.05);compared with that before occlusion,there was significant difference from 45 min after occlusion in EA group(P<0.05);compared with that before occlusion,there was significant difference from 105 min after occlusion in sham group(P<0.05);there was significant difference at 120 min after occlusion comparing with 60 min after occlusion in EA group;compared with sham group,there was significant difference at every time point in EA group and Is group(except 30 min after occlusion)(P<0.05),respectively;compared with Is group,there was significant difference at 30,45,60 min after occlusion in EA group(P<0.05).④The infarct brain volume detected 24 h after reperfusion in EA group was significantly smaller comparing with that in Is group(P<0.05).Conclusion:Electro-acupuncture preconditioning could increase CBF velocity in artery and vein of focal cortex in rats with MCAO,which also had time-dependence.Additionally,electro-acupuncture preconditioning could decrease the infarct volume detected 24h after reperfusion,which possibly was correlated with the blood supply in ischemic penumbra.展开更多
OBJECTIVE To investigate the neuroprotective effect and possible mechanisms of lute.olin-7-O-β-D-glucuronide(LGU) against focalcerebral ischemic injury.METHODS The focal cerebral ischemic injury model was established...OBJECTIVE To investigate the neuroprotective effect and possible mechanisms of lute.olin-7-O-β-D-glucuronide(LGU) against focalcerebral ischemic injury.METHODS The focal cerebral ischemic injury model was established by middle cerebral artery occlusion(MCAO).Male Sprague Dawley rats were randomly divided into sham group,model group(MCAO),LGU group(0.24,0.72 and2.16 mg·kg^(-1)) and positive control group(Edaravone at 5 mg·kg^(-1)).LGU was injected intravenously 30 min after MCAO.Neurological severity score,infarct volume and brain water content were detected 24 h after MCAO and the levels of Na+-K+ ATPase,Ca2 + ATPase,TNF-α and IL-1β were detected to explore the possible mechanisms.For the therapeutic time window test,LGU(0.72 mg·kg^(-1)) was injected intrave.nously 0.5,2,4,6,8,10 and 12 h respectively after MCAO.To evaluate motion behavior,LGU were injected intravenously 30 min after MCAO and once per day during detection period.The changes of motor coordination were detected by rotating rod method and grip strength analysis,and the changes of gaits were detected using DigiGait Imaging System.RESULTS LGU improved the neurological severity score,infarct volume ratio and brain water content.The therapeutic time window of LGU for cerebral infarction and brain edema was at least 6 h and for neurological dysfunction was 12 h.LGU also prolonged the latency on rotarod,increased the forelimb tension and improved 8 gait parameters,including stance duration,stride length,stance width,paw area,paw area variability,gait symmetry,ataxia coefficient and tau propulsion.Furthermore,LGU increased Na^+-K^+-ATPase and Ca^(2+)-ATPase levels in the cortex and hippocampus in the ischemic side,reduced the levels of TNF-α and IL-1β in the serum.CONCLUSION LGU has a significant neuroprotective effect against cerebral ischemic injury via improving energy metabolism and reducing inflammation.展开更多
OBJECTIVE To study the treatment effect of Gastrodia elata and Uncaria rhynchophylla water extract(GUW)against cerebral ischemia,and the metabolomic profile of cerebrospinal fluid in GUW treated cerebral ischemia in r...OBJECTIVE To study the treatment effect of Gastrodia elata and Uncaria rhynchophylla water extract(GUW)against cerebral ischemia,and the metabolomic profile of cerebrospinal fluid in GUW treated cerebral ischemia in rat model.METHODS Middle cerebral artery occlusion was performed in male SD rat.GUW or control(PBS)was administered orally for 7d post-operationally.At day 7,the brain of the rat was harvested.The infarct volume of the brain was measured after TTC staining.The brain was subjected to H&E and cresyl violet staining in histological analysis.The cerebrospinal fluid(CSF)of another set of animals was aspirated.The CSF samples were labeled with 13 C-dansyl and 12C-dansyl chloride.The reaction mixture was put into liquid chromatography Fourier-transform ion cyclotron resonance mass spectrometry for analysis of the CSF metabolome.Multivariate statistical analysis will be carried out to determine the differences of metabolomic profile in CSF of GUW versus control group.RESULTS After GUW treatment,the neurological deficit score was significantly improved from day 3 to 7(P<0.001).GUW treated group significantly reduced 55.7%infarct volume on MCAO rats in TTC stained brain slices(P<0.001)and preserved brain histological structure observed in H&E and cresyl violet staining.On the other hand,according to various statistical analyses to compare CSF metabolite in the GUW and control group,volcano plot analysis showed 48 metabolites(P<0.001);OPLS-DA analysis showed 129 metabolites(VIP score >2.0)and 45 metabolites in ROC curve analysis.Among identified metabolites,L-aspartic acid(60%)and glycine(62%)decreased in GUW while threoninyl-alanine increased by 1.6 fold significantly.CONCLUSION GUW ameliorated cerebral ischemic injury,and the metabolomic approach may be a potential means to find biomarker of the Chinese medicine treatment on cerebral ischemia.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The project supported by Hunan province Science and Technology Plan Projects of China(2015DK3010)
文摘OBJECTIVE To study the role of Ginkgo biloba extract-761(EGb-761)in the recovery of gait abnormality and its neuroprotective effect against the brain injury induced by permanent middle cerebral artery occlu-sion in rats.METHODS Male Sprague Dawley rats(n=200,240-305 g)were anesthetized with 0.2%pentobarbital sodium diluted in physiological saline(2.0 m L·kg-1,ip).Then a monofilament coated with poly-L-lysine,was used to occlude the origin of the middle cerebral artery.It was inserted into the internal carotid artery lumen until it met mild resistance,approximately 20mm beyond the common carotid artery bifurcation.The suture was secured with a ligature and maintained in place until sacrifice.The same surgical procedure was conducted in sham-operated rats in which the middle cerebral artery was not occluded.Motor and behavioral changes were assessed after surgery using a five point scale.The rats securing the point scale above 2 were included in the study.The rats were randomly divided into control,and treated groups:EGb-761(20,50,and 100 mg·kg-1).The treated groups were oral y administered(10 mL·kg-1)for 28 d.On 7th,14th,21st,and 28th day the neurological scores,rotar rod test and gait assessment(the automated computer-assisted method)were performed.The brains were collected for TTC staining and histopathological analysis.RESULTS 1)Weight:On 28th day,EGb-761(20 mg·kg-1,)significantly increased the weight of the rat by^8%as compared to control(~300 g).However,at 50 mg·kg-1,and 100 mg·kg-1,a significant increase of^7-7.6%(control:~232 g),and^7.3-7%,respectively from 14 to 28 days was noted.2)Neurological scores:On 28thday,EGb-761(20,50,and 100 mg·kg-1)significantly decreased the neurological scores by^18%,~22%,~21%,respectively as compared to control(~2.07).3)Rotar rod test:On 28thday,EGb-761(50,and100 mg·kg-1)significantly increased by^69.1%,~74.1%,respectively as compared to control(~28.2).4)Gait assessment:On 7th,14th,21st,and 28thday,EGb-761(20,50,and 100 m·kg-1)significantly reduced the average body angle,on 7th,14th,21st,and 28thday,EGb-761(100 mg·kg-1)significantly increased the walk speed and reduced the average walking cycle,EGb-761(50,and 100 mg·kg-1)significantly the area of the left brain/right brain area percentage and reduced tissue pathologic neuron injury.CONCLUSION Ginkgo biloba extract EGb-761 has obvious improve behavior disorders,and has a protective neuroprotective effect against the brain injury induced by permanent middle cerebral artery occlusion.
基金supported by the Natural Science Foundation of China(NSFC,NO.81171856,81171855)the Development Program of China(863 program,NO.2007AA02Z482)the Key projects of Shanghai Science and Technology on Biomedicine(NO.10DZ1950800)
文摘Objective:To investigate the effects of electro-acupuncture preconditioning on focal cerebral blood flow(CBF)in artery and vein of rats with middle cerebral artery occlusion(MCAO),and to provide experimental evidences for primary prevention of ischemic stroke.Method:Eighteen male Sprague-Dawley rats(two months old)were divided into 3 groups:electro-acupuncture preconditioning group(EA group),ischemia group(Is group)and Dazhui and Baihui sham group(six rats per group).The rats in the EA group were given electro-acupuncture preconditioning at Dazhui and Baihui with 2/15 density wave current for 30 minutes per day,5 days consecutively.After preconditioning,enhanced laser speckle contrast analysis was implemented to get false color images before making middle cerebral ischemia occlusion model.Then getting false color images two hours during ischemia by laser speckle contrast analysis again.Result:④Relative CBF in focal cortical artery:There were significant differences at every time point in EA group and Is group(P<0.01,P<0.05)comparing with that before occlusion respectively.Compared with that before ischemia,there was significant difference at 120 min after ischemia in sham group(P<0.05);compared with that at 30 min after occlusion,there was significant difference at every time point in EA group from the time point 60 min after occlusion;there was significant difference at every time point in EA group and Is group(P<0.05)comparing with sham group;Compared with sham group,there was significant difference at 30min and 60 min after ischemia in EA group(P<0.05).④Relative CBF in focal cortical vein:There was significant difference at every time point in sham group comparing with that before occlusion(P<0.05);compared with that before occlusion,there was significant difference from 45 min after occlusion in EA group(P<0.05);compared with that before occlusion,there was significant difference from 105 min after occlusion in sham group(P<0.05);there was significant difference at 120 min after occlusion comparing with 60 min after occlusion in EA group;compared with sham group,there was significant difference at every time point in EA group and Is group(except 30 min after occlusion)(P<0.05),respectively;compared with Is group,there was significant difference at 30,45,60 min after occlusion in EA group(P<0.05).④The infarct brain volume detected 24 h after reperfusion in EA group was significantly smaller comparing with that in Is group(P<0.05).Conclusion:Electro-acupuncture preconditioning could increase CBF velocity in artery and vein of focal cortex in rats with MCAO,which also had time-dependence.Additionally,electro-acupuncture preconditioning could decrease the infarct volume detected 24h after reperfusion,which possibly was correlated with the blood supply in ischemic penumbra.
基金supported by Young and Middle-aged Teacher Career Development Support Plan of Shenyang Pharmaceutical University(ZQN2016002) and Science and Technology Funds from Department of Education of Liaoning province(2016101633L3)
文摘OBJECTIVE To investigate the neuroprotective effect and possible mechanisms of lute.olin-7-O-β-D-glucuronide(LGU) against focalcerebral ischemic injury.METHODS The focal cerebral ischemic injury model was established by middle cerebral artery occlusion(MCAO).Male Sprague Dawley rats were randomly divided into sham group,model group(MCAO),LGU group(0.24,0.72 and2.16 mg·kg^(-1)) and positive control group(Edaravone at 5 mg·kg^(-1)).LGU was injected intravenously 30 min after MCAO.Neurological severity score,infarct volume and brain water content were detected 24 h after MCAO and the levels of Na+-K+ ATPase,Ca2 + ATPase,TNF-α and IL-1β were detected to explore the possible mechanisms.For the therapeutic time window test,LGU(0.72 mg·kg^(-1)) was injected intrave.nously 0.5,2,4,6,8,10 and 12 h respectively after MCAO.To evaluate motion behavior,LGU were injected intravenously 30 min after MCAO and once per day during detection period.The changes of motor coordination were detected by rotating rod method and grip strength analysis,and the changes of gaits were detected using DigiGait Imaging System.RESULTS LGU improved the neurological severity score,infarct volume ratio and brain water content.The therapeutic time window of LGU for cerebral infarction and brain edema was at least 6 h and for neurological dysfunction was 12 h.LGU also prolonged the latency on rotarod,increased the forelimb tension and improved 8 gait parameters,including stance duration,stride length,stance width,paw area,paw area variability,gait symmetry,ataxia coefficient and tau propulsion.Furthermore,LGU increased Na^+-K^+-ATPase and Ca^(2+)-ATPase levels in the cortex and hippocampus in the ischemic side,reduced the levels of TNF-α and IL-1β in the serum.CONCLUSION LGU has a significant neuroprotective effect against cerebral ischemic injury via improving energy metabolism and reducing inflammation.
基金The project supported by the Health and Medical Research Fund,Hong Kong(11120381)
文摘OBJECTIVE To study the treatment effect of Gastrodia elata and Uncaria rhynchophylla water extract(GUW)against cerebral ischemia,and the metabolomic profile of cerebrospinal fluid in GUW treated cerebral ischemia in rat model.METHODS Middle cerebral artery occlusion was performed in male SD rat.GUW or control(PBS)was administered orally for 7d post-operationally.At day 7,the brain of the rat was harvested.The infarct volume of the brain was measured after TTC staining.The brain was subjected to H&E and cresyl violet staining in histological analysis.The cerebrospinal fluid(CSF)of another set of animals was aspirated.The CSF samples were labeled with 13 C-dansyl and 12C-dansyl chloride.The reaction mixture was put into liquid chromatography Fourier-transform ion cyclotron resonance mass spectrometry for analysis of the CSF metabolome.Multivariate statistical analysis will be carried out to determine the differences of metabolomic profile in CSF of GUW versus control group.RESULTS After GUW treatment,the neurological deficit score was significantly improved from day 3 to 7(P<0.001).GUW treated group significantly reduced 55.7%infarct volume on MCAO rats in TTC stained brain slices(P<0.001)and preserved brain histological structure observed in H&E and cresyl violet staining.On the other hand,according to various statistical analyses to compare CSF metabolite in the GUW and control group,volcano plot analysis showed 48 metabolites(P<0.001);OPLS-DA analysis showed 129 metabolites(VIP score >2.0)and 45 metabolites in ROC curve analysis.Among identified metabolites,L-aspartic acid(60%)and glycine(62%)decreased in GUW while threoninyl-alanine increased by 1.6 fold significantly.CONCLUSION GUW ameliorated cerebral ischemic injury,and the metabolomic approach may be a potential means to find biomarker of the Chinese medicine treatment on cerebral ischemia.
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 the Natural Science Foundation of Hunan Province(2020JJ4865 and S2021JJMSXM2689)the Hunan Provincial Clinical Medical Technology Innovation Guidance Project(2020SK53612),China.
