目的探讨快速老化小鼠SAMP8老年性痴呆的关键细胞学机制。方法以2月龄和8月龄SAMP8小鼠各40只为痴呆相关快速老化动物模型,以同月龄各40只正常老化小鼠SAMR1为对照,从小鼠海马组织提取脂筏蛋白,采用高效液相色谱-串联质谱法分析。脂筏...目的探讨快速老化小鼠SAMP8老年性痴呆的关键细胞学机制。方法以2月龄和8月龄SAMP8小鼠各40只为痴呆相关快速老化动物模型,以同月龄各40只正常老化小鼠SAMR1为对照,从小鼠海马组织提取脂筏蛋白,采用高效液相色谱-串联质谱法分析。脂筏蛋白质组学检测数据导入DAVID生物信息学分析工具,进行GeneOntology(GO)生物信息学分析和Kyoto Encyclopedia of Genes and Genomes(KEGG)代谢网络分析,并用线粒体膜电位和Morris水迷宫方法验证生物信息学分析结果。结果与SAMR1小鼠比较,快速老化的SAMP8小鼠出现明显的认知障碍。GO分析显示,老年期SAMP8小鼠脂筏蛋白组中线粒体相关蛋白大幅度减少。KEGG分析显示,老年期SAMP8小鼠海马组织线粒体的氧化磷酸化功能大幅度衰退。线粒体膜电位分析显示,老年期SAMP8小鼠海马组织线粒体膜电位大幅度降低。结论在老化过程中,SAMP8小鼠海马组织最关键的细胞变化是线粒体氧化磷酸化功能的过度衰退,这可能是其痴呆发生的重要细胞学机制。展开更多
Objective To explore the effect of electroacupuncture(EA)intervention on the vasoconstriction of cerebral artery smooth muscle cells after cerebral infarction.Methods Male Wistar rats were randomly divided into 3 grou...Objective To explore the effect of electroacupuncture(EA)intervention on the vasoconstriction of cerebral artery smooth muscle cells after cerebral infarction.Methods Male Wistar rats were randomly divided into 3 groups by a random number table:the model group(n=24),the EA group(n=24),and the normal group(n=6).The model and the EA groups were divided into different time subgroups at 0.5,1,3,and 6 h after middle cerebral artery occlusion(MCAO),with 6 rats in each subgroup.MCAO model was established using intraluminal suture occlusion method.The EA group was given EA treatment at acupoint Shuigou(GV 26)instantly after MCAO for 20 min.The contents of cerebrovascular smooth muscle MLCK,the 3 subunits of myosin light chain phosphatase(MLCP)MYPT1,PP1c-δand M20,as well as myosin-ATPase activity were detected using immunohistochemistry and Western blotting.Results The overall expression level of the MYPT1 and PP1c-δin the model group was significantly higher(P<0.01).After EA intervention,the 0.5 h group expression level was close to that of the normal group(P>0.05),and the other subgroups were still significantly higher than the normal group(P<0.01).After EA intervention,the expression level of each subgroup was significantly lower than the corresponding model group.There was a significant difference between the 0.5 and 1 h subgroups(P<0.01),while a difference was also observed between the 3 and 6 h subgroups(P<0.05).The dynamic change rule gradually increased with the prolongation of infarction time within 6 h after infarction.Conclusion EA intervention can inhibit contraction of cerebral vascular smooth muscle cells and regulate smooth muscle relaxation by regulating MLCK pathway.展开更多
Objective:To investigate the effect of electro-acupuncture(EA)on vasomotor symptoms in rats with acute cerebral infarction,by observing the changes in the expression of factors related to the phosphatidylinositol(PI)s...Objective:To investigate the effect of electro-acupuncture(EA)on vasomotor symptoms in rats with acute cerebral infarction,by observing the changes in the expression of factors related to the phosphatidylinositol(PI)system.Methods:Forty-two Wistar rats were randomly divided into 3 groups by a random number table:the control group(n=6),the model group(n=18)and the EA group(n=18).The EA group was given EA treatment at Shuigou(GV 26)instantly after modeling with middle cerebral artery occlusion(MCAO)method,while the model and control groups were not given any treatment.The degrees of neurological deficiency were evaluated using neurological severity scores(NSS)and the brain blood flow was evaluated by a laser scanning confocal microscope.Western blot analysis was conducted to detect the expression levels of G-protein subtype(Gq)and calmodulin(CaM).Competition for protein binding was conducted to detect the expression level of inositol triphosphate(IP3).Thin layer quantitative analysis was conducted to detect the expression level of diacylglycerol(DAG).The expression level of intracellular concentration of free calcium ion([Ca^(2+)]i)was detected by flow cytometry.Results:The NSS of the model group was significantly higher than the control group at 3 and 6 h after MCAO(P<0.01),while the EA group was significantly lower than the model group at 6 h(P<0.01).The cerebral blood flow in the model group was significantly lower than the control group at 1,3 and 6 h after MCAO(P<0.01),while for the EA group it was remarkably higher than the model group at the same time points(P<0.01).The expressions of Gq,CaM,IP3,DAG and[Ca^(2+)]i in the model group were significantly higher than the control group(P<0.05 or P<0.01),and those in the EA group were significantly lower than the model group at the same time points(P<0.05 or P<0.01).Conclusion:EA treatment at GV 26 can effectively decrease the over-expression of related factors of PI system in rats with acute cerebral infarction,improve cerebral autonomy movement,and alleviate cerebral vascular spasm.展开更多
文摘目的探讨快速老化小鼠SAMP8老年性痴呆的关键细胞学机制。方法以2月龄和8月龄SAMP8小鼠各40只为痴呆相关快速老化动物模型,以同月龄各40只正常老化小鼠SAMR1为对照,从小鼠海马组织提取脂筏蛋白,采用高效液相色谱-串联质谱法分析。脂筏蛋白质组学检测数据导入DAVID生物信息学分析工具,进行GeneOntology(GO)生物信息学分析和Kyoto Encyclopedia of Genes and Genomes(KEGG)代谢网络分析,并用线粒体膜电位和Morris水迷宫方法验证生物信息学分析结果。结果与SAMR1小鼠比较,快速老化的SAMP8小鼠出现明显的认知障碍。GO分析显示,老年期SAMP8小鼠脂筏蛋白组中线粒体相关蛋白大幅度减少。KEGG分析显示,老年期SAMP8小鼠海马组织线粒体的氧化磷酸化功能大幅度衰退。线粒体膜电位分析显示,老年期SAMP8小鼠海马组织线粒体膜电位大幅度降低。结论在老化过程中,SAMP8小鼠海马组织最关键的细胞变化是线粒体氧化磷酸化功能的过度衰退,这可能是其痴呆发生的重要细胞学机制。
基金Supported by the National Natural Science Foundation of China(Nos.81674056 and 82074543)the Tianjin Municipal Natural Science Foundation(No.18JCYBJC94200),China。
文摘Objective To explore the effect of electroacupuncture(EA)intervention on the vasoconstriction of cerebral artery smooth muscle cells after cerebral infarction.Methods Male Wistar rats were randomly divided into 3 groups by a random number table:the model group(n=24),the EA group(n=24),and the normal group(n=6).The model and the EA groups were divided into different time subgroups at 0.5,1,3,and 6 h after middle cerebral artery occlusion(MCAO),with 6 rats in each subgroup.MCAO model was established using intraluminal suture occlusion method.The EA group was given EA treatment at acupoint Shuigou(GV 26)instantly after MCAO for 20 min.The contents of cerebrovascular smooth muscle MLCK,the 3 subunits of myosin light chain phosphatase(MLCP)MYPT1,PP1c-δand M20,as well as myosin-ATPase activity were detected using immunohistochemistry and Western blotting.Results The overall expression level of the MYPT1 and PP1c-δin the model group was significantly higher(P<0.01).After EA intervention,the 0.5 h group expression level was close to that of the normal group(P>0.05),and the other subgroups were still significantly higher than the normal group(P<0.01).After EA intervention,the expression level of each subgroup was significantly lower than the corresponding model group.There was a significant difference between the 0.5 and 1 h subgroups(P<0.01),while a difference was also observed between the 3 and 6 h subgroups(P<0.05).The dynamic change rule gradually increased with the prolongation of infarction time within 6 h after infarction.Conclusion EA intervention can inhibit contraction of cerebral vascular smooth muscle cells and regulate smooth muscle relaxation by regulating MLCK pathway.
基金the National Natural Science Foundation of China(Nos.82074543,81473765,81674056)the Tianjin Municipal Natural Science Foundation(No.18JCYBJC94200)。
文摘Objective:To investigate the effect of electro-acupuncture(EA)on vasomotor symptoms in rats with acute cerebral infarction,by observing the changes in the expression of factors related to the phosphatidylinositol(PI)system.Methods:Forty-two Wistar rats were randomly divided into 3 groups by a random number table:the control group(n=6),the model group(n=18)and the EA group(n=18).The EA group was given EA treatment at Shuigou(GV 26)instantly after modeling with middle cerebral artery occlusion(MCAO)method,while the model and control groups were not given any treatment.The degrees of neurological deficiency were evaluated using neurological severity scores(NSS)and the brain blood flow was evaluated by a laser scanning confocal microscope.Western blot analysis was conducted to detect the expression levels of G-protein subtype(Gq)and calmodulin(CaM).Competition for protein binding was conducted to detect the expression level of inositol triphosphate(IP3).Thin layer quantitative analysis was conducted to detect the expression level of diacylglycerol(DAG).The expression level of intracellular concentration of free calcium ion([Ca^(2+)]i)was detected by flow cytometry.Results:The NSS of the model group was significantly higher than the control group at 3 and 6 h after MCAO(P<0.01),while the EA group was significantly lower than the model group at 6 h(P<0.01).The cerebral blood flow in the model group was significantly lower than the control group at 1,3 and 6 h after MCAO(P<0.01),while for the EA group it was remarkably higher than the model group at the same time points(P<0.01).The expressions of Gq,CaM,IP3,DAG and[Ca^(2+)]i in the model group were significantly higher than the control group(P<0.05 or P<0.01),and those in the EA group were significantly lower than the model group at the same time points(P<0.05 or P<0.01).Conclusion:EA treatment at GV 26 can effectively decrease the over-expression of related factors of PI system in rats with acute cerebral infarction,improve cerebral autonomy movement,and alleviate cerebral vascular spasm.
