粉防己碱对心肌成纤维细胞增殖、活化的影响  

Effects of tetrandrine on proliferation and activation of cardiac fibroblasts

作　　者：李乾 常亮[2] 苏冬梅 马旭 LI Qian;CHANG Liang;SU Dong-mei;MA Xu(Human Genetic Resources Center,National Research Institute for Family Planning,Beijing 100081,China;Department of Obstetrics and Gynecology,Peking University Third Hospital,Beijing 100191,China)

机构地区：[1]国家卫生计生委科学技术研究所人类遗传资源中心,北京100081 [2]北京大学第三医院妇产科生殖医学中心,北京100191

出　　处：《北京大学学报（医学版）》2018年第2期331-334,共4页Journal of Peking University:Health Sciences

基　　金：国家自然科学基金(31301145)资助~~

摘　　要：目的:探讨粉防己碱(tetrandrine,Tet)对大鼠心肌成纤维细胞增殖、活化的影响。方法:采用细胞计数试剂盒(cell counting kit-8C,CCK-8)的方法检测不同浓度Tet对心肌成纤维细胞增殖的抑制作用;采用蛋白质印迹(Western blot)法检测Tet对由转化生长因子β(transforming growth factor-β,TGF-β)诱导的心肌成纤维细胞β-链蛋白(β-catenin)、波形蛋白(vimentin,Vm)、纤连蛋白(fibronectin,Fn)及平滑肌α-肌动蛋白(smooth muscleα-actin,SMA)的表达上调的影响作用;采用实时荧光定量PCR的方法检测Tet作用下,TGF-β诱导的心肌成纤维细胞Ⅰ型胶原(collagen-1,Col-1)和Ⅲ型胶原(collagen-3,Col-3)的表达上调的变化。结果:CCK-8检测结果显示,分别用0.5、1、2、4、8μmol/L的Tet处理心肌成纤维细胞,其增殖活力分别下降至对照组的94.4%、84.9%、74.9%、63.8%、50.3%,差异有统计学意义(所有P值<0.05;1、2、4、8μmol/L处理组P值<0.001)。Western blot检测结果显示:5μg/L的TGF-β显著上调β-catenin、Vm、Fn及SMA的表达(P值分别为0.001、0.008、0.010、0.001),而用0.5μmol/L的Tet预处理心肌成纤维细胞后,可以阻断由TGF-β诱导的β-catenin、Fn、SMA的表达上调(P值分别为0.009、0.005、0.019),但不改变Vm的表达变化(P>0.05)。荧光定量PCR检测结果表明:用0.5μmol/L的Tet预处理心肌成纤维细胞,可以减弱由TGF-β诱导的Ⅰ型及Ⅲ型胶原的表达上调,差异有统计学意义(P<0.001)。结论:Tet可以显著抑制心肌成纤维细胞的增殖,并阻断由TGF-β诱导的心肌成纤维细胞活化,据此推断,Tet可能具有抗心肌纤维化的作用,进而阻断心肌重塑。Objective:To investigate the effects of tetrandrine(Tet)on proliferation and activation of rat cardiac fibroblasts.Methods:Firstly,the cell counting kit-8(cck-8)assay was applied to detect the effects of Tet with different concentrations on proliferation of cardiac fibroblasts.Secondly,transforming growth factor(TGF-β)with a concentration of 5μg/L was used to induce the cardiac fibroblast activation,and Western blot was performed to measure the expression variation ofβ-catenin,vimentin(Vm),fibronectin(Fn)and smooth muscleα-actin(SMA).At last,the real-time PCR was conducted to measure the expression change of collagen-1(Col-1)and collagen-3(Col-3).Results:The cck-8 assay showed that the Tet with different concentrations respectively,which were 0.5μmol/L,1μmol/L,2μmol/L,4μmol/L,and 8μmol/L,significantly inhibited the proliferation of cardiac fibroblasts.The viability was decreased to 94.4%,84.9%,74.9%,63.8%and 50.3%respectively of the control group when the Tet concentration changed,and the difference was statistically significant,P=0.043,P<0.001,P<0.001,P<0.001,P<0.001 respectively.Western blot revealed that the expressions ofβ-catenin,Fn,SMA and Vm,were up-regulated by TGF-β(5μg/L),the result showed that the difference was statistically significant,and the P values were 0.001,0.008,0.010,0.001 respectively.Then,the up-regulation ofβ-catenin,Fn and SMA was attenuated by pre-treatment of Tet,and the result also displayed that the difference was statistically significant,and the P values were 0.009,0.005,0.019,respectively.While there was no significant change in the expression of Vm,according to Western blotting,and P>0.05,at the same time,real-time PCR indicated that the up-regulations of Col-1 and Col-3 which were induced by TGF-βwere blocked by pre-treatment of Tet,the result showed that the difference was statistically significant,P<0.001.Conclusion:According to the experimental results,we can draw the conclusion that:the Tet can significantly inhibit the proliferation of cardiac fibroblasts,meanw

关 键 词：粉防己碱 心肌成纤维细胞 心肌重塑 

分 类 号：R965[医药卫生—药理学]