电针水沟穴对脑缺血再灌注大鼠神经细胞自噬的影响

发布时间：2018-01-12 07:23

  本文关键词：电针水沟穴对脑缺血再灌注大鼠神经细胞自噬的影响　出处：《第二军医大学》2016年硕士论文　论文类型：学位论文

  更多相关文章： 电针 水沟穴 脑缺血再灌注 自噬 LC3B p62 Bcl-2

【摘要】：目的:探讨电针水沟穴对MCAO/R大鼠神经细胞自噬的影响及其可能的作用机制。方法:本实验参照Longa法并改良制备大鼠大脑中动脉缺血再灌注(MCAO/R)模型,大鼠脑缺血2h后再灌注24h和72h。将雄性SD大鼠随机分为假手术组、模型组和治疗组,每组按时相分为24h和72h 2个亚组。运用Bederson评分法对各组大鼠进行神经功能评分。于再灌注后24h和72h 2个时间点取材,TTC染色法观察并计算各组大鼠脑梗死体积比;光镜观察各组大鼠神经细胞形态结构;透射电镜观察神经细胞自噬体情况;western blot法检测各组大鼠自噬相关蛋白LC3B、p62的表达;免疫组化法检测Bcl-2阳性细胞的表达并计算Bcl-2阳性细胞率。结果:1.与同时相假手术组比较,模型24h组和模型72h组Bederson评分升高(p0.01),脑梗死体积比上升(p0.05),神经细胞病理损害加重。治疗24h组和治疗72h组较同时相模型组Bederson评分降低(p0.05),梗死体积比减小(p0.05),神经细胞损伤减轻。2.模型24h组透射电镜下观察到神经细胞自噬体数量较多;与模型24h组比较,治疗24h组神经细胞自噬体数量减少。模型72h组和治疗72h组未观察到自噬体。3.模型24h组和模型72h组LC3B-II/LC3B-I的比值较假手术24h组升高(p0.01)。治疗24h组和治疗72h组LC3B-II/LC3B-I的比值与同时相模型组比较无统计学差异(p0.05)。4.模型24h组p62表达较假手术24h组降低(p0.05);模型72h组p62表达与假手术24h组比较无统计学差异(p0.05)。治疗24h组p62表达较模型24h组升高(p0.05);治疗72h组p62表达与模型72h组比较无统计学差异(p0.05)。5.与假手术24h组比较,模型24h组和模型72h组Bcl-2阳性细胞表达率升高,差异有统计学意义(p0.01);治疗24h组Bcl-2阳性细胞表达率较模型24h组升高(p0.01),治疗72h组Bcl-2阳性细胞表达率与模型72h组比较无统计学差异(p0.05)。结论:1.电针水沟穴缩小脑缺血再灌注损伤大鼠脑梗死体积,促进脑缺血再灌注损伤大鼠神经功能恢复,对脑缺血再灌注损伤大鼠起到脑保护作用。2.电针水沟穴下调神经细胞自噬水平可能是其对脑缺血再灌注损伤大鼠发挥脑保护作用的机制之一。3.电针水沟穴减轻脑缺血再灌注损伤大鼠神经细胞自噬可能与其上调p62、Bcl-2表达水平有关。
[Abstract]:Objective: to investigate the effect of electroacupuncture at Shuigou point on neuronal autophagy in MCAO/R rats and its possible mechanism. MCAO / R) model. Male Sprague-Dawley rats were randomly divided into sham-operation group, model group and treatment group. Each group was divided into 2 subgroups at 24 h and 72 h. The neurological function of rats in each group was evaluated by Bederson scoring method. The samples were collected at 24 and 72 hours after reperfusion. The volume ratio of cerebral infarction was observed and calculated by TTC staining. The morphologic structure of nerve cells in each group was observed by light microscope. The neuronal autophagy was observed by transmission electron microscope. The expression of autophagy associated protein LC3BP62 was detected by western blot. Immunohistochemical method was used to detect the expression of Bcl-2 positive cells and calculate the rate of Bcl-2 positive cells. The Bederson scores of 24 h model group and 72 h model group were increased (p0.01), and the cerebral infarction volume ratio was increased (p0.05). Compared with the model group, the Bederson score and the infarct volume ratio of the 24 h and 72 h groups were lower than those of the model group (P 0.05 and P 0.05 respectively). The number of neuronal autophagy was observed under transmission electron microscope in the model group. The results were compared with the model group for 24 hours. The number of neuronal autophagy decreased in 24 h treatment group. No autophagy was observed in model group and 72 h group. The ratio of LC3B-II/LC3B-I in model group 24 h and model group 72 h was not observed. The value was higher than that in the 24 h sham operation group (. The ratio of LC3B-II/LC3B-I in 24 h group and 72 h group was not significantly different from that in the simultaneous model group (p 0.05). The expression of p62 in the 24 h model group was lower than that in the sham operation 24 h group. There was no significant difference in the expression of p62 between the model group and the sham operation group (24 h), but the expression of p62 in the 24 h treatment group was higher than that in the 24 h model group. There was no significant difference in p62 expression between the 72 h treatment group and the model 72 h group, and there was no significant difference between the treatment group and the sham operation 24 h group. The expression rate of Bcl-2 positive cells in 24 h model group and 72 h model group was higher than that in model 24 h group and 72 h group, the difference was statistically significant (P 0.01). The expression rate of Bcl-2 positive cells in the treatment group was higher than that in the model group (P 0.01). There was no significant difference in the expression rate of Bcl-2 positive cells between the 72 h group and the model group (P 0.05). Conclusion: electroacupuncture at Shuigou point can reduce the volume of cerebral infarction in rats with cerebral ischemia-reperfusion injury. To promote the recovery of nerve function in rats with cerebral ischemia-reperfusion injury. Electroacupuncture at Shuigou point down-regulating the level of neuronal autophagy may be one of the mechanisms of its protective effect on cerebral ischemia-reperfusion injury in rats. The effect of Acupoint on neuronal autophagy in rats with cerebral ischemia-reperfusion injury may be related to its up-regulation of p62. The expression level of Bcl-2 is related.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R245.97

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