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
目的探讨急性脑缺血/再灌注大鼠神经元细胞骨架蛋白时空动态变化。方法线栓法阻断大鼠大脑中动脉90 min后实现再灌注,在不同再灌注时间点观察及取材。尼氏染色观察神经细胞损伤,采用神经功能缺损评分和前肢放置实验评估神经功能;免疫组...目的探讨急性脑缺血/再灌注大鼠神经元细胞骨架蛋白时空动态变化。方法线栓法阻断大鼠大脑中动脉90 min后实现再灌注,在不同再灌注时间点观察及取材。尼氏染色观察神经细胞损伤,采用神经功能缺损评分和前肢放置实验评估神经功能;免疫组化染色、免疫印迹法观察细胞骨架成分微管相关蛋白2(microtubule associated protein 2,MAP2)、神经丝重链(neurofilament heavy chain,NF-H)的变化;透射电镜观察轴突、树突和神经丝亚显微结构。结果随着再灌注时间的延长,脑损伤和神经行为功能损害逐渐加重。纹状体损伤比皮层出现的更早、更严重。缺血区域的MAP2相关免疫反应强度降低,NF-H相关免疫反应强度升高。超微结构观察显示细胞骨架排列受损,密度降低。结论不同脑区对缺血/再灌注损伤的耐受性不同。神经元细胞骨架的主要成分对缺血和再灌注表现出动态反应,这可能进一步促进脑损伤和神经功能缺损。展开更多
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.
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.
文摘目的探讨急性脑缺血/再灌注大鼠神经元细胞骨架蛋白时空动态变化。方法线栓法阻断大鼠大脑中动脉90 min后实现再灌注,在不同再灌注时间点观察及取材。尼氏染色观察神经细胞损伤,采用神经功能缺损评分和前肢放置实验评估神经功能;免疫组化染色、免疫印迹法观察细胞骨架成分微管相关蛋白2(microtubule associated protein 2,MAP2)、神经丝重链(neurofilament heavy chain,NF-H)的变化;透射电镜观察轴突、树突和神经丝亚显微结构。结果随着再灌注时间的延长,脑损伤和神经行为功能损害逐渐加重。纹状体损伤比皮层出现的更早、更严重。缺血区域的MAP2相关免疫反应强度降低,NF-H相关免疫反应强度升高。超微结构观察显示细胞骨架排列受损,密度降低。结论不同脑区对缺血/再灌注损伤的耐受性不同。神经元细胞骨架的主要成分对缺血和再灌注表现出动态反应,这可能进一步促进脑损伤和神经功能缺损。
