血脂康对新生大鼠心肌成纤维细胞增殖及胶原合成的影响

目的观察血脂康对血管紧张素Ⅱ(AngⅡ)诱导的新生大鼠心肌成纤维细胞(CFs)增殖及胶原合成的影响,并探讨可能的分子机制。方法采用胰酶消化法分离培养Sprague-Dawley大鼠心肌成纤维细胞,建立AngⅡ诱导CFs增殖的模型,以MTT比色法和流式细胞仪检测细胞周期分析法观察血脂康对CFs数目和细胞周期的影响。AngⅡ及不同浓度血脂康作用48 h后,用天狼星红染色法检测培养上清中胶原的含量;ELISA法检测细胞培养上清液中TGF-β1蛋白表达;RT-PCR法检测胶原和TGF-β1 mRNA表达。结果AngⅡ对CFs增殖有明显促进作用,血脂康可明显抑制AngⅡ诱导的CFs增殖,且呈剂量依赖性;血脂康可增加G0/G1期细胞百分率,降低S、G2/M期细胞百分率,降低胶原含量、TGF-β1蛋白及胶原和TGF-β1 mRNA的表达。结论血脂康能抑制AngⅡ诱导的CFs增殖和胶原的产生,其作用可能是通过抑制TGF-β1表达实现的。

Effect of Blood Fat Regulator Xuezhikang on Proliferation and Collagen Synthesis of Cardiac Fibroblast of Newborn Rats

Objective To observe the effect of Xuezhikang,an extract of cholestin,on the proliferation and collagen synthesis of cardiac fibroblasts(CFs) caused by angiotensin Ⅱ(AngⅡ) in newborn rats and explore the potential molecular mechanism of the effect.Methods Isolate the CFs of newborn Sprague-Dawley rats by trypsin digestion method,and induce the proliferation of CFs by AngⅡ.Observe the effect of Xuezhikang on the number and cell cycle of CFs by MTT method and flow cytometry.After treatment with AngⅡ and Xuezhikang at various concentrations for 48 h,the yield of collagen in culture supernatant was determined by Sirius staining,and the expression of TGF-β1 protein by ELISA.The expressions of collagen gene and TGF-β1 mRNA were determined by RT-PCR.Results AngⅡ promoted the proliferation of CFs significantly.However,Xuezhikang showed significant dose-dependent inhibitory effect on proliferation of CFs induced by AngⅡ.The drug increased the percentage of cells at G0/G1 phase,decreased those at S and G2/M phases,and decreased the expressions of collagen gene and TGF-β1 mRNA.Conclusion Xuezhikang inhibited the proliferation of CFs and production of collagen induced by AngⅡ.The mechanism for this might be the inhibition of TGF-β1 expression.