乌头碱对H9c2细胞分化的影响  

Effects of aconitine on H9c2 cell differentiation

作　　者：张斌强 陈龙 冉凤英 杜开慧 余飞 陈炜 杨丽萍 薛旸 陈琴华 陈继舜 ZHANG Binqiang;CHEN Long;RAN Fengying;DU Kaihui;YU Fei;CHEN Wei;YANG Liping;XUE Yang;CHEN Qinhua;CHEN Jishun(Medical Experimental Center,Affiliated Dongfeng Hospital,Hubei University of Medicine,Shiyan,Hubei 442008,China;Department of Cardiovascular Medicine,Affiliated Dongfeng Hospital,Hubei University of Medicine,Shiyan,Hubei 442008,China)

机构地区：[1]湖北医药学院附属东风医院实验中心,湖北十堰442008 [2]湖北医药学院附属东风医院心血管内科,湖北十堰442008

出　　处：《安徽医药》2020年第7期1287-1290,I0001,共5页Anhui Medical and Pharmaceutical Journal

基　　金：湖北省教育厅科学研究计划指导性项目(B2018435、B2016140);湖北省卫生健康科研基金资助(WJ2019M054);十堰市科学技术研究与开发项目(17Y49、17Y53、17Y54)。

摘　　要：目的观察乌头碱对H9c2细胞分化的影响。方法用不同浓度的乌头碱作用于大鼠H9c2心肌细胞5 d后噻唑蓝比色法(MTT)检测细胞的增殖率。选取50μM/L的乌头碱作用于全反式维甲酸(RA)诱导分化体系下的大鼠H9c2心肌细胞。光学显微镜观察细胞形态变化,荧光定量PCR(q PCR)检测actc1、myl1、myl2、tnnt2、nppb的mRNA表达,免疫荧光细胞化学染色检测α肌动蛋白(α-actinin)分布情况。蛋白质印迹法(Western Blot)检测α-actinin、心肌肌钙蛋白I(cTnI)蛋白的表达。采用单因素方差分析对比组间差异。结果乌头碱能够呈剂量依赖性的抑制H9c2心肌细胞增殖(P<0.05)。较分化模型组,乌头碱处理组中分化细胞形态减少,actc1[(2.30±0.17)比(0.55±0.03),F=62.17、P=0.000],myl1[(2.57±0.15)比(1.22±0.13),F=63.15、P=0.000],myl2[(18.64±0.53)(10.02±1.10),F=206.65、P=0.000],tnnt2[(5.53±0.23)(2.23±0.10),F=216.89、P=0.000]的mRNA表达降低(P<0.01)差异有统计学意义,cTnI及α-actinin的蛋白表达降低。结论乌头碱能够抑制H9c2心肌细胞分化。Objective To observe the influence of aconitine on the H9 c2 cells differentiation.MethodsThe proliferation rate of rats’H9 c2 cardiomyocytes was detected by thiazolyl blue colorimetric method(MTT)treated with different concentrations of aconitine after 5 days.50μM/L aconitine was selected to act on H9 c2 under the induced differentiation system of all-trans Retinoic Acid(RA).The morphological changes of cells were observed by optical microscopy.The mRNA expression of actc1,myl1,myl2,tnnt2 and nppb was detected by fluorescence quantitative PCR(qPCR),and distribution of cardiacα-actinin was detected by immunofluorescence cytochemical staining.The expressions of cardiac troponin I(cTnI)andα-actinin protein were detected by Western-Blot.Statistical analysis was performed by One-way analysis of variance to compare differences between groups.ResultsAconitine can inhibit the proliferation of H9 c2 in a concentration-dependent manner.Aconitine can inhibit the mRNA expression ofactc1[(2.30±0.17)vs.(0.55±0.03),F=62.17、P=0.000],myl1[(2.57±0.15)vs.(1.22±0.13),F=63.15、P=0.000],myl2[(18.64±0.53)vs.(10.02±1.10),F=206.65、P=0.000],tnnt2[(5.53±0.23)vs.(2.23±0.10),F=216.89、P=0.000]and the proteins expression of c TnI andα-actinin.ConclusionAconitine can inhibit differentiation of H9 c2 cardiomyocytes.

关 键 词：乌头碱 肌细胞 心脏 细胞分化 H9C2 肌钙蛋白I Α肌动蛋白 

分 类 号：R285[医药卫生—中药学]