复元醒脑汤对糖尿病脑梗死大鼠microRNA-503及CCNE1、CDC25A蛋白表达的影响及对促进血管新生的实验研究  

Effect of Fuyuan Xingnao Decoction on Expression of MicroRNA-503 and CDC25A、CCNE1 And its Experimental Study on Promoting Angiogenesis in Diabetic Rats with Cerebral Infarction

作　　者：沈俊逸 方邦江[2] 赵智明[1] 刘春丽[1] 壮雨雯 徐文俊 蔡辉[1] Shen Junyi;Fang Bangjiang;Zhao Zhiming(Nanjing General Hospital of Nanjing Military Command,Jiangsu,Nanjing 210002,China.)

机构地区：[1]南京军区南京总医院,江苏南京210002 [2]上海中医药大学附属龙华医院,上海200032

出　　处：《中国中医急症》2018年第11期1881-1885,1890,共6页Journal of Emergency in Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81273725)

摘　　要：目的观察复元醒脑汤对microRNA-503及CCNE1、CDC25A蛋白表达的影响,探讨复元醒脑汤促进糖尿病脑梗死血管新生的分子机制。方法 150只健康雄性SD大鼠随机分为空白对照组、假手术组、模型组、西药治疗组、中药治疗组,每组30只。采用腹腔注射链脲佐菌素(STZ)后予以高能量高脂肪饲料喂养2周的方法复制糖尿病大鼠模型,制模后采用线栓法制备糖尿病脑梗死动物模型。完成造模后,中药治疗组灌胃复元醒脑汤;西药治疗组灌胃二甲双胍、尼莫地平;空白对照组、假手术组、模型组均灌胃等量的0.9%氯化钠注射液。各组连续灌胃14 d后处死取材。免疫荧光法检测脑梗死区域的血管新生、微循环观察仪检测脑梗死区域血管微循环状况、实时荧光定量PCR法检测脑组织microRNA-503、CCNE1、CDC25A基因表达水平,利用Western blotting检测CCNE1、CDC25A蛋白表达水平。结果与模型组相比,中药治疗组和西药治疗组的脑梗死区域都出现血管新生增多,血管管径增粗,血流加快,蛋白渗漏减少,少见白细胞黏附现象且中药治疗组优于西药治疗组;各实验组脑组织的CCNE1、CDC25A基因及蛋白表达水平由高到低依次为:空白对照组、假手术组、中药治疗组、西药治疗组、模型组。各实验组脑组织microRNA-503基因的表达水平由低到高依次是:空白对照组、假手术组、中药治疗组、西药治疗组、模型组,中药治疗组和西药治疗组均显著低于模型组(P <0.01)。结论复元醒脑汤通过上调CCNE1、CDC25A的表达,从而抑制microRNA-503的表达水平,达到促进梗死组织区域的血管新生,改善脑部微循环。Objective： To observe the effect of Fuyuan Xingnao Decoction on the expression of mi R-503,CCNE1 and CDC25 A protein,to explore the molecular mechanism of Fuyuan Xingnao Decoction in promoting angiogenesis in diabetic cerebral infarction. Methods： 150 healthy male SD rats were divided into two groups： control group,sham operation group,model group,western medicine treatment group and traditional Chinese medicine treatment group,with 30 rats in each group. After intraperitoneal injection of STZ the rats were fed with high energy and high fat diet for 2 weeks to replicate the model of diabetic rats. The cerebral infarction model was made by line plug method. In Chinese medicine group were treated with Fuyuan Xingnao Decoction. Western medicine treatment group were treated with metformin. Model group,sham operation group were given the same amount of normal saline for 14 days. After the last time of gastric gavage,blood vessel of cerebral infarction area was detected by immunofluorescence. Microcirculation of cerebral infarction area was detected by microcirculation observation instrument.The gene expression of brain tissue CCNE1,CDC25 A,microRNA-503 was detected by Real time fluorescent quantitative PCR method,and CCNE1,CDC25 A protein expression level by was detected by Western blotting. Results： Compared with Model group,the angiogenesis in the cerebral infarction area of western medicine treatment group and traditional Chinese medicine treatment group risen;the diameter of blood vessel was increased;blood flow has accelerated;the leakage of protein reduced;and leukocyte adhesion phenomenon was rare. Meanwhile,the traditional Chinese medicine treatment group was better than western medicine treatment group. The CCNE1,CDC25 A gene and protein expression in each experimental group had the same trend,from high to low as follows： control group,sham operation group,Chinese medicine treatment group,western medicine treatment group,model group,and traditional. The brain tissue microRNA-503 gene expression expr

关 键 词：糖尿病脑梗死 脑部微循环 复元醒脑汤 血管新生 microRNA-503 

分 类 号：R285.5[医药卫生—中药学]