黄精地龙提取液对老年痴呆大鼠脑内烟碱型乙酰胆碱受体的影响  

Effect of Extracts from Polygonati Rhizoma and Pheretima on Nicotinic Acetylcholine Receptor in Brain of Senile Dementia Rats

作　　者：肖移生[1] 高萌[1] 侯吉华[1] 

机构地区：[1]江西中医药大学基础医学院(生命科学学院),南昌330004

出　　处：《中国实验方剂学杂志》2017年第14期140-145,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：江西省教育厅2015年度科技项目(GJJ150868)

摘　　要：目的:探讨黄精地龙提取液(EPP)对老年痴呆大鼠脑内α3和α7烟碱型乙酰胆碱受体(α3,α7-nAChR)阳性神经元数量、蛋白及mRNA表达的影响。方法:SD大鼠40只,随机均分为正常组,模型组,脑复康(阳性药物,0.6 g·kg^(-1)·d^(-1))组,EPP(相当生药量4 g·kg^(-1)·d^(-1))组。给大鼠颈背部皮下注射1%D-半乳糖(5 m L·kg^(-1)·d^(-1),连续3周)后,腹腔注射东莨菪碱(2 mg·kg^(-1)·d^(-1),连续后2周)复制大鼠老年痴呆模型;造模1周后同时对脑复康组及EPP组大鼠每天灌胃给相应实验药物,正常组及模型组大鼠灌胃给生理盐水1 mL。实验结束,处死各组大鼠,取出脑组织,一半置于10%中性甲醛液浸泡固定待组织切片检查,另一半置于冰上匀浆,进行mRNA及蛋白提取;运用尼氏染色、免疫组化观察各大鼠大脑皮层第一躯体感觉区(S1Tr)和海马CA1与CA3区的阳性神经元数量;实时荧光定量聚合酶链式反应(Real-time PCR),蛋白质免疫印迹(Western blot)检测各大鼠大脑内的α3和α7-nAChR mRNA及蛋白的表达。结果:与正常组比较,模型组大鼠大脑S1Tr区和海马CA1及CA3区神经元数量显著减少,大脑内α3和α7-n AChR mRNA和蛋白表达水平均降低(P<0.01);EPP能增加痴呆大鼠大脑S1Tr区和海马CA1及CA3区神经元数量,能升高痴呆大鼠大脑内α3和α7-n AChR mRNA和蛋白的表达水平,与模型组比较差异有统计学意义(P<0.05,P<0.01)。结论:EPP能保护大脑认知学习记忆有关重要区的神经细胞,同时触发α3和α7-n AChR受体效应,提高中枢乙酰胆碱(ACh)含量,进而升高老年痴呆大鼠中枢胆碱能系统活性。Objective： To observe the effects of Extracts from Polygonati Rhizoma and Pheretima （EPP） on the number of α3 and α7-nieotinic acid type aeetyleholine receptors （nAChR） neurons, the expression of proteins and mRNA in brains of senile dementia rats. Method： The 40 SD rats were randomly divided into normal group, model group, piraeetam （ positive control drug, 0.6 g·kg^-1·d^-1 ） group, and EPP （ equivalent raw drug dose, 4 g·kg^-1·d^-1） group. The senile dementia models were established by nuchal subcutaneous injection of 1% D-galactose （ 5 mL·kg^-1·d^-1, 3 consecutive weeks） and intraperitoneal injection of Scopolamine （ 2 mg·kg^-1·d^-1, 2 consecutive weeks） in rats. 1 week after modeling, the rats in piracetam group and EPP group received corresponding drugs daily by intragastric administration, and the rats in control group and model group were given with normal saline 1 mL. At the end of the experiment, all the rats were sacrificed, and their brain tissues weretaken out. Half of the brain tissues was fixed by 10% neutral formaldehyde solution for biopsy, and the other half was homogenated on ice to extract mRNA and protein. The count of positive neurons in hippocampal CA1, CA3 area and S1Tr areas were detected by Nissl staining and immunohistochemistry methods; The α3 and α7-nAChR mRNA and protein expression levels in the brains of senile dementia rats were measured by Real-time PCR and Western blot respectively. Result： As compared with the normal group, the counts of neurons in hippocampal CA1 and CA3 area and S1Tr area were decreased, and the expression levels of α3 and α7-nAChR mRNA and proteins in the brains were also decreased in model group （P 〈 0.01 ）; EPP could increase the count of neurons in hippocampal CA1 and CA3 area and S1Tr area; EPP could increase the expression of α3 and α7-nAChR mRNA and protein in the brains of dementia rats as compared with the model group （P 〈 0.05, P 〈 0.01 ）. Conclusion ： EPP can protect the nerve cells in the

关 键 词：黄精地龙提取液 老年痴呆大鼠 大脑皮层 海马 神经元 烟碱型乙酰胆碱受体(nAChR) 

分 类 号：R285.5[医药卫生—中药学]