摘要
目的探讨鸢尾苷元对血管紧张素Ⅱ(AngⅡ)作用下心肌成纤维细胞增殖和凋亡的影响及其机制。方法采用差速贴壁法获得纯化的大鼠心肌成纤维细胞,采用MTT法和原位末端标记法检测鸢尾苷元或磷脂肌醇-3-激酶(PI3K)/蛋白激酶B(Akt)信号通路对AngⅡ作用下心肌成纤维细胞增殖和凋亡的影响,采用RT-PCR检测鸢尾苷元对AngⅡ作用下心肌成纤维细胞中凋亡相关基因Bcl-2、Bax mRNA表达的影响,Western blot法检测鸢尾苷元对AngⅡ作用下心肌成纤维细胞中凋亡相关蛋白Bcl-2、Bax和PI3K/Akt信号通路相关蛋白p-Akt、T-Akt表达的影响。结果AngⅡ能明显促进心肌成纤维细胞增殖,抑制细胞凋亡;给予50,100和200μmol·L^-1鸢尾苷元后,AngⅡ使心肌成纤维细胞促增殖和抑凋亡作用明显受到抑制,且200μmol·L^-1鸢尾苷元作用最显著。200μmol·L^-1鸢尾苷元能够明显抑制AngⅡ诱导的Bcl-2 mRNA和p-Akt、Bcl-2蛋白的表达;减弱AngⅡ对Bcl-2 mRNA和蛋白表达的抑制作用,不影响T-Akt蛋白表达。PI3K/Akt信号通路抑制剂LY294002作用后,AngⅡ使心肌成纤维细胞的促增殖和抑凋亡作用明显受到抑制;而给予PI3K/Akt信号通路激活剂HY-N1412作用后,AngⅡ对心肌成纤维细胞的促增殖和抑凋亡作用明显得到促进,HY-N1412能明显逆转鸢尾苷元对AngⅡ刺激下的心肌成纤维细胞抑增殖和促凋亡作用。结论鸢尾苷元可通过PI3K/Akt信号通路减弱AngⅡ对心肌成纤维细胞促增殖和抑凋亡作用。
Objective To investigate the effects of iridin on proliferation and apoptosis of cardiac fibroblasts under AngⅡand its mechanism.Methods Rat cardiac fibroblasts were purified by differential adhesion method.The effects of iritin or PI3K/Akt signaling pathway on the proliferation and apoptosis of cardiac fibroblasts induced by AngⅡwere detected by MTT and in situ end labeling.The effect of iritin on the expression of apoptosis-related genes Bcl-2 and Bax mRNA in cardiac fibroblasts induced by AngⅡwas detected by RT-PCR.The effect of iritin on the expression of P-Akt and T-Akt proteins related to PI3K/Akt signaling pathway in myocardial fibroblasts induced by AngⅡwas detected by Western blot.Results The proliferation of cardiac fibroblasts could be promoted obviously and the apoptosis of cells could be inhibited by AngⅡ.The effect of promoting proliferation and apoptosis inhibition of AngⅡon cardiac fibroblasts was obviously inhibited after treated with 50,100 and 200μmol·L^-1 iritin,and the effect of 200μmol·L^-1 iridin was the most significant.The expression of Bcl-2 mRNA and p-Akt,Bcl-2 proteins induced by AngⅡcould be inhibited significantly,while the inhibitory effect of AngⅡon Bcl-2 mRNA and protein expression was weakened by 200μmol·L^-1 iritin,but the expression of T-Akt protein could not be affected.The effect of promoting proliferation and apoptosis inhibition of AngⅡon cardiac fibroblasts was obviously inhibited after treated with PI3K/Akt signaling pathway inhibitor LY294002,while the effect of promoting proliferation and apoptosis inhibition of AngⅡon cardiac fibroblasts was significantly promoted after the PI3K/Akt signaling pathway activator HY-N1412 acted,and the effects of iridin on proliferation and apoptosis of cardiac fibroblasts stimulated by AngⅡwere reversed by HY-N1412.Conclusion Iridogen can attenuate the effect of AngⅡon proliferation and apoptosis of cardiac fibroblasts through PI3K/Akt signaling pathway.
作者
陈雪斌
汪凤兰
张冠茂
CHEN Xuebin;WANG Fenglan;ZHANG Guanmao(Department of Cardiology,Puyang People′s Hospital,Puyang 457000,China)
出处
《西北药学杂志》
CAS
2020年第1期62-66,共5页
Northwest Pharmaceutical Journal
基金
河南省科技攻关计划项目(编号:182102310142)
