补肾填精益髓法对脑出血模型大鼠神经保护作用及Notch-1蛋白表达的影响  

Influences of Bushen Tianjing Yisui Method on Neuroprotective Effect and Expression of Notch-1 Protein in Intracerebral Hemorrhage Rats

作　　者：张毅 魏绪旺 许康 张昭 薛瑶瑶 范天祥 ZHANG Yi;WEI Xuwang;XU Kang;ZHANG Zhao;XUE Yaoyao;FAN Tianxiang(The Affiliated Hospital of Shaanxi University of Traditional Chinese Medicine,Xianyang 712000,Shaanxi,China)

机构地区：[1]陕西中医药大学附属医院,陕西咸阳712000 [2]陕西中医药大学第一临床医学院

出　　处：《中西医结合心脑血管病杂志》2020年第5期751-756,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：陕西省中医药管理局基金资助项目(No.JCPT-025)。

摘　　要：目的探讨补肾填精益髓法对脑出血(ICH)大鼠神经保护作用及Notch-1蛋白表达的影响,明确其具体作用机制。方法将120只SD大鼠随机分为5组:假手术组、模型组、补肾填精益髓组、神经节苷脂组、γ-分泌酶抑制剂(DAPT)组。采用尾动脉注血法制作脑出血大鼠模型,5-溴脱氧尿嘧啶核苷(BrdU)腹腔注射标记脑内增殖细胞,在第3天、第7天、第14天、第28天4个时间点采用免疫组织化学法标记大鼠海马区BrdU/神经元核抗原(NeuN)阳性细胞数;尼色染色法观察大鼠脑出血灶周围组织病理形态结构;Werstern Blot法测定Notch-1蛋白表达情况。结果术后第7天、第14天、第28天,补肾填精益髓组、神经节苷脂组BrdU/NeuN阳性细胞数均高于模型组、DAPT组(P<0.01),且补肾填精益髓组BrdU/NeuN阳性细胞数均高于神经节苷脂组(P<0.01),以第7天、第14天增殖最明显,随后呈下降趋势;术后第7天、第14天,补肾填精益髓组大鼠Bederson神经功能评分较模型组、神经节苷脂组、DAPT组改善明显(P<0.01),至术后第28天Bederson神经功能评分基本降至正常;补肾填精益髓组和神经节苷脂组Notch-1蛋白表达高于模型组、DAPT组(P<0.01);在第7天、第14天时,补肾填精益髓组与神经节苷脂组Notch-1蛋白表达上升最明显,且补肾填精益髓组Notch-1蛋白表达高于神经节苷脂组表达(P<0.01)。结论补肾填精益髓法可以改善脑出血大鼠神经功能,促进神经干细胞(NSCs)增殖,其机制可能是补肾填精益髓法可通过Notch信号通路诱导尾状核出血区域NSCs增殖,提高Notch-1蛋白表达。Objective To investigate the effect of Bushen Tianjing Yisui method on neuroprotective effect and expression of Notch-1 protein in intracerebral hemorrhage(ICH)rats,and to clarify its specific mechanism.Methods One hundred and twenty SD rats were randomly divided into 4 groups:sham operation group,model group,Bushen Tianjing Yisui group,ganglioside group andγ-secretase inhibitor(DAPT)group.The model of ICH was made by tail-artery blood-injection,BrdU was used to label the proliferating cells in the brain,and the number of BrdU/NeuN positive cells in the hippocampus of rats was marked by immunocytochemical method at 3,7,14,and 28 days.The expression of Notch-1 protein was determined by Werstern Blot method.Results On the 7th,14th,and 28th day after operation,the number of BrdU/NeuN positive cells in Bushen Tianjing Yisui group and ganglioside group were higher than those in the model group and DAPT group(P<0.01).The number of BrdU/NeuN positive cells in Bushen Tianjing Yisui group was higher than that of ganglioside group at each time point(P<0.01),and the proliferation was the most obvious at the 7th and 14th day,and then decreased.On the 7th,14th,28th day after operation,the Bederson scores of rats in Bushen Tianjing Yisui group were significantly improved than that in model group,ganglioside group,and DAPT group(P<0.01),and the neurological function score was basically reduced to normal on the 28th day after operation.The expression of Notch-1 protein in Bushen Tianjing Yisui group and ganglioside group were higher than those in model group and DAPT group(P<0.01).On the 7th and 14th day,the expression of Notch-1 protein increased most significantly in the Bu shen Tianjing Yisui group and ganglioside group,and the expression of Notch-1 protein in Bushen Tianjing Yisui group was higher than that in the ganglioside group(P<0.01).Conclusion Bushen Tianjing Yisui method can improve the nerve function,promote the proliferation of neural stem cells(NSCs)in ICH rats.The mechanism may be that Bushen Tianjing Yisui meth

关 键 词：脑出血 补肾填精益髓法 神经干细胞 神经节苷脂 Notch-1蛋白 

分 类 号：R743[医药卫生—神经病学与精神病学] R285.5[医药卫生—临床医学]