补阳还五汤对动脉粥样硬化大鼠血管平滑肌细胞NF-κB p65 mRNA及NF-κB p65蛋白表达量的影响  

Effects of Buyanghuanwu decoction on the expression of rho kinase and NF-κB p65 and NF-κB p65 protein in atherosclerotic rats

作　　者：郭春兰 钟秉知 刘建春 王青 姜维佳 宋丽娟 张红珍 Guo Chunlan;Zhong Bingzhi;Liu Jianchun;Wang Qing;Jiang Weijia;Song Lijuan;Zhang Hongzhen(The Fourth Clinical Medical College,Shanxi University of Chinese Medicine,Jinzhong Shanxi 030619;Northwest A&F University,Xianyang Shannxi 712100;Biological Research Center of Shanxi University of Chinese Medicine,The Key Laboratory Yiqi Huoxue of State Administration of Traditional Chinese Medicine,Jinzhong Shanxi 030619)

机构地区：[1]山西中医药大学第四临床学院,山西晋中030619 [2]西北农林科技大学,陕西咸阳712100 [3]山西中医药大学生物学研究中心,国家中医药管理局多发性硬化益气活血重点研究室,山西晋中030619

出　　处：《山西中医药大学学报》2021年第6期405-409,共5页Journal of Shanxi University of Chinese Medicine

基　　金：国家自然科学基金项目(81373532);山西省黄芪资源产业化及产业国际化协同创新中心项目(HQXTCXZX2016-024)。

摘　　要：目的:观察补阳还五汤对动脉粥样硬化(atherosclerosis,AS)大鼠主动脉NF-κB p65 mRNA及NF-κB p65蛋白表达量的影响。方法:取正常SD大鼠30只,随机分为正常对照组、补阳还五汤低剂量对照组[10 g/(kg·d)]和补阳还五汤高剂量对照组[20 g/(kg·d)];另取造模成功的AS模型大鼠30只,随机分为AS模型组、补阳还五汤低剂量治疗组[10 g/(kg·d)]和补阳还五汤高剂量治疗组[10 g/(kg·d)];正常对照组、AS模型组每日给予等体积生理盐水灌胃,连续灌胃10 d。第10天制备含药血清。将平滑肌细胞放于不同含药血清中培养,用4种不同药物进行干预,48 h后取主动脉提取RNA并进行逆转录,将cDNA样品稀释8倍作为模板上机进行PCR检测。6种不同血清及不同药物处理后,NF-κB p65蛋白提取及浓度采用Western Blot方法测定。结果:各AS模型组含药血清作用后NF-κB p65mRNA的表达水平明显高于正常对照组;与AS模型组相比,空白对照组的补阳还五汤低剂量治疗组、高剂量治疗组,阳性对照组的补阳还五汤低剂量治疗组、高剂量治疗组,ROCK激动剂组的补阳还五汤高剂量组、低剂量治疗组,ROCK抑制剂组的补阳还五汤高剂量治疗组药血清作用后NF-κB p65mRNA的表达水平明显降低。补阳还五汤高剂量治疗组血管平滑肌细胞NF-κB p65蛋白的表达均低于AS模型组。结论:补阳还五汤可能通过下调NF-κB p65mRNA及NF-κB p65蛋白的表达减轻血管平滑肌细胞的增殖,起到抗动脉粥样硬化作用。Objective:To study the effect of Buyanghuanwu decoction on expression of NF-κB p65 mRNA and NF-κB p65 protein in atherosclerotic rats. Method:Normal 30 rats were randomly divided into three groups:the normal control group,Buyanghuanwu decoction low dose of control group [10 g/(kg·d)]、Buyanghuanwu decoction high dose of control group 20 g/(kg·d);successful AS model 30 rats were randomly divided into three groups:the model group,Buyanghuanwu decoction low dose of therapy group[10 g/(kg·d)]、Buyanghuanwu decoction high dose of therapy group[20 g/(kg·d)].The normal control group and AS model group were given equal volume normal saline daily for 10 days. RNA was extracted from aorta and retrieved. The DNA sample was diluted 8 times as a template for PCR. Western blot was used to determine the concentration and extraction of NF-κB p65 protein from 6 different serum samples. Results:The expression level of NF-κB p65 mRNA in drug-containing serum of AS model control group was significantly higher than that of normal control group. Compared with AS model control group,blank control group was treated with low-dose and high-dose Buyanghuanwu decoction;positive control group was treated with low-dose and high-dose Buyanghuanwu decoction,ROCK agonist group was treated with high-dose and low-dose Buyanghuanwu decoction. The expression level of NF-κB p65 mRNA in ROCK inhibitor group was significantly decreased after treatment with high dose of Buyanghuanwu decoction. The expression of NF-κB p65 protein in vascular smooth muscle cells of Buyanghuanwu decoction group was lower than that of AS model control group. Conclusion:BuyangHuanwu decoction may has an effect of resisting atherosclerosis by reduce the proliferation of vascular smooth muscle cells by down-regulating the expression of NF-kappa B p65 and NF-κB P65 protein.

关 键 词：补阳还五汤 动脉粥样硬化 NF-κB p65 

分 类 号：R285[医药卫生—中药学]