黄连素对冠心病大鼠心肌细胞凋亡的影响及其机制研究  

Influences of Berberine on Cardiomyocyte Apoptosis in Coronary Heart Disease Rats and its Mechanism

作　　者：曹雪明 朱娜 CAO Xueming;ZHU Na(Henan Provincial People′s Hospital,Zhengzhou 450000,Henan,China)

机构地区：[1]河南省人民医院

出　　处：《中西医结合心脑血管病杂志》2019年第22期3504-3507,共4页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：国家自然科学基金(No.U1704167)

摘　　要：目的研究黄连素(BBR)对冠心病大鼠心肌细胞凋亡的影响及其作用机制。方法以高脂饮食喂养并注射脑垂体后叶素建立冠心病大鼠模型。将模型动物分为实验组和对照组,同时以正常SD大鼠设立空白组,实验组予以高、中、低剂量黄连素灌胃,对照组和空白组予以等黄连素体积的双蒸水灌胃,治疗周期为12周。酶联免疫吸附法(ELISA)和硝酸酶还原法检测血清肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)含量;实时荧光定量(RT-qPCR)检测Calpain1和Calpain2 mRNA表达水平;蛋白印记实验检测Cleaved-Caspase 3、Bcl-2、Bax、Calpain1及Calpastatin相对蛋白表达量。结果①与空白组相比,对照组TNF-α、IL-1β、IL-6含量均显著上升(P<0.05);与对照组相比,3种剂量组TNF-α、IL-1β、IL-6含量均有不同程度下降(P<0.05)。②与空白组相比,对照组Calpain1 mRNA相对水平明显升高(P<0.01),Calpain2 mRNA无明显变化;与对照组相比,高、中、低剂量实验组Calpain1 mRNA相对水平呈剂量依赖性降低,差异有统计学意义(P<0.05或P<0.01),而Calpain2无明显变化。③与空白组相比,对照组大鼠心肌组织Calpain1蛋白表达量增多(P<0.01),而Calpastatin蛋白表达明显下降(P<0.01);与对照组相比,实验组大鼠心肌组织Calpastatin蛋白表达明显增多(P<0.05或P<0.01),而Calpain1蛋白表达显著下降(P<0.01),并呈剂量依赖性。④与空白组相比,对照组大鼠心肌组织Bcl-2蛋白表达量明显降低(P<0.01),而Bax和Cleaved-Caspase 3表达升高(P<0.01);与对照组相比,实验组大鼠心肌组织Bcl-2蛋白表达增多,Bax和Cleaved-Caspase 3蛋白表达明显下降,并呈剂量依赖性,差异均有统计学意义(P<0.05或P<0.01)。结论黄连素能降低冠心病大鼠心肌细胞凋亡,该作用可能与增强Calpain1蛋白表达和活性,下调血清中炎症因子TNF-α、IL-1β、IL-6水平有关。Objective To study the effect of berberine(BBR) on myocardial cell apoptosis in rats with coronary heart disease and its mechanism.Methods Rats model of coronary heart disease was established by feeding with a high-fat diet and injection of pituitrin.The model rats were divided into test group and control group.At the same time,normal SD rats were used to establish blank group.The test group was given high-,middle-,and low-dose BBR gavage.The control group and the blank group were treated with distilled water of BBR volume.The treatment cycle was 12 weeks.Serum TNF-α,IL-1β,and IL-6 levels were measured by ELISA and nitrate reduction test;Calpain1 and Calpain2 mRNA expression levels were detected by RT-qPCR;Cleaved-Caspase 3,Bcl-2,Bax,Calpain1,and Calpastatin relative protein expressions were detected by western blotting.Results ①Compared with the blank group,the levels of TNF-α,IL-1β,and IL-6 in the control group increased significantly(P<0.05);compared with the control group,the levels of TNF-α,IL-1β,and IL-6 in the three test group decreased to different extents(P<0.05).②Compared with the blank group,the relative level of Calpain1 mRNA in the control group was significantly higher(P<0.01),and there was no significant change in Calpain2 mRNA.Compared with the control group,the relative levels of Calpain1 mRNA in high-,medium-,and low-dose test groups decreased by a dose-dependent manner,and the difference was statistically significant(P<0.05 or P<0.01),but there was no significant change in Calpain 2.③Compared with the blank group,the expression of Calpain1 protein in the experimental group was increased(P<0.01),but Calpastatin protein expression was significantly decreased;Compared with the control group,the expression of Calpastatin protein in the myocardial tissue of high-,medium-,low-dose test group was significantly increased(P<0.05 or P<0.01),while the expression of Calpain1 was significantly decreased by a dose-dependent manner(P<0.01).The Results were statistically significant(P<0.05).④Comp

关 键 词：冠心病 黄连素 心肌 炎症因子 细胞凋亡 实验研究 

分 类 号：R541.4[医药卫生—心血管疾病] R256.2[医药卫生—内科学]