野黄芩苷对血管紧张素Ⅱ诱导的新生大鼠心肌成纤维细胞的增殖及ERK1/2、p38 MAPK信号通路的影响

Effects of Scutellarin on the Proliferation of Cardiac Fibroblasts in Neonate Rats Induced by Angiotensin Ⅱ and Signal Pathway of ERK1/2 and p38 MAPK

目的:研究野黄芩苷对血管紧张素Ⅱ(AngⅡ)诱导的新生大鼠心肌成纤维细胞(CFs)的增殖及细胞外调节蛋白激酶(ERK1/2)、p38丝裂原活化蛋白激酶(p38 MAPK)信号通路的影响。方法:体外分离培养新生大鼠CFs,将细胞分为空白组(空白培养基)、AngⅡ组(10^(-7)μmol/L)、50μmol/L野黄芩苷组和AngⅡ(10^(-7)μmol/L)+5、10、20、50μmol/L野黄芩苷组,培养48 h后,采用CCK-8法检测细胞增殖能力。另取细胞分为空白组(空白培养基)、AngⅡ组(10^(-7)μmol/L)和AngⅡ(10^(-7)μmol/L)+5、10、20、50μmol/L野黄芩苷组,培养48 h后,检测细胞中Ⅰ型胶原(ColⅠ)、ColⅢ、α-平滑肌肌蛋白(α-SMA)m RNA表达和细胞培养液中羟脯氨酸(HYP)含量,以及细胞中ERK1/2、p38 MAPK磷酸化水平。结果:5、10、20、50μmol/L的野黄芩苷均能显著抑制AngⅡ诱导的CFs的增殖(P<0.05),显著下调AngⅡ刺激的CFs中ColⅠ、ColⅢ和α-SMA m RNA的表达(P<0.05),显著抑制AngⅡ刺激后细胞培养液中HYP含量的增加和细胞中ERK1/2、p38 MAPK的磷酸化(P<0.05),且具有一定的浓度依赖性。结论:野黄芩苷对AngⅡ诱导的CFs的增殖具有一定的抑制作用,其机制可能与抑制ERK1/2、p38 MAPK的磷酸化有关。

OBJECTIVE:To study the effects of scutellarin on the proliferation of cardiac fibroblasts(CFs)in neonate rats induced by angiotensinⅡ(ANGⅡ)and signal pathway of extracellular regulated protein kinase(ERK1/2)and p38 mitogen activated protein kinase(p38 MAPK).METHODS:CFs of neonatal rat were isolated and cultured in vitro,and then divided into blank group(blank culture medium),AngⅡgroup(10-7μmol/L),50μmol/L scutellarin group and AngⅡ(10-7μmol/L)+5,10,20,50μmol/L scutellarin groups.After cultured for 48 h,CCK-8 assay was used to detect the proliferation ability of cells.Other cells were selected and divided into blank group(blank culture medium),AngⅡgroup(10-7μmol/L)and AngⅡ+5,10,20,50μmol/L scutellarin groups.After cultured for 48 h,mRNA expression of ColⅠ,ColⅢandα-SMA in cells and hydroxyproline(HYP)content in cell culture fluid were detected;phosphorylation levels of ERK1/2 and p38 MAPK in cells were also detected.RESULTS:5,10,20,50μmol/L scutellarin could significantly inhibit the proliferation of CFs induced by AngⅡ(P<0.05),and decreased mRNA expression of ColⅠ,ColⅢandα-SMA in CFs induced AngⅡ(P<0.05).5,10,20,50μmol/L scutellarin could significantly inhibit the increase of HYP content and the phosphorylation of ERK1/2 and p38 MAPK after induced by AngⅡ(P<0.05),in dose-dependent manner.CONCLUSIONS:Scutellarin inhibits the proliferation of CFs induced by AngⅡ,the mechanism of which may be associated with reduction of ERK1/2 and p38 MAPK phosphorylation.