摘要
目的:研究血府逐瘀汤对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)诱导的心肌成纤维细胞增殖及细胞外基质合成的影响,探讨其抑制心肌纤维化可能的作用机制。方法:采用胰酶消化法分离培养乳鼠心肌成纤维细胞(cardiac fibroblast,CF),建立AngⅡ诱导的心肌纤维化模型。实验分组为:空白对照组,AngⅡ组,5%、10%、15%血府逐瘀汤组和缬沙坦组。空白对照组以对照组鼠血清温育,其余各组先以AngⅡ10-6mol/L刺激24h后,再分别以对照组鼠血清,5%、10%、15%血府逐瘀汤含药血清,缬沙坦含药血清温育24h。采用四氮唑蓝比色法检测CF增殖,羟脯氨酸法检测胶原含量,放射免疫法检测细胞培养上清中透明质酸(hy-aluronic acid,HA)和Ⅲ型前胶原(procollagenⅢ,PCⅢ)水平,酶联免疫吸附测定法检测上清中纤维连接蛋白(fibronectin,FN)表达。结果:与空白对照组比较,AngⅡ10-6mol/L作用CF24h显著上调CF增殖指数和DNA合成(P<0.01),增加胶原含量(P<0.01),增加上清液中FN、HA和PCⅢ含量(P<0.01)。与AngⅡ10-6mol/L相比,血府逐瘀汤含药血清下调CF增殖指数和DNA合成,降低胶原含量(P<0.05),减少上清液中FN、HA和PCⅢ含量(P<0.01)。结论:AngⅡ能够促进CF增殖及细胞外基质合成,具有促进心肌纤维化作用;血府逐瘀汤能改善心肌纤维化,机制可能与抑制AngⅡ诱导CF增殖,抑制细胞外基质胶原蛋白、HA、PCⅢ及FN合成有关。
Objective：To investigate the effects of Xuefu Zhuyu Decoction,a compound Chinese herbal medicine,on the proliferation and extracellular matrix synthesis of cardiac fibroblasts（CFs）from neonatal rats induced by angiotensin Ⅱ,and to explore the mechanism of Xuefu Zhuyu Decoction in improving myocardial fibrosis.Methods：CFs derived from neonatal rats were cultured with enzymatic dissociation to establish a cell model of cardiac fibrosis induced by angiotensin Ⅱ in vitro.CFs were divided into 6 groups,including blank control group,angiotensin Ⅱ group,valsartan-medicated serum group,and 5%,10% and 15% Xuefu Zhuyu decoction-medicated serum group.CFs in the blank control group were incubated with normal rat serum;CFs in the other groups were stimulated with angiotensin Ⅱ at a dose of 10-6 mol/L,and then incubated with normal rat serum and 5%,10% and 15% Xuefu Zhuyu decoction-medicated serum for 24 h,respectively.Methyl thiazolyl tetrazolium assay was employed to measure CF proliferation,and collagen synthesis was detected by analysis of the hydroxyproline in CFs.Radioimmunoassay was used to measure concentrations of procollagen Ⅲ and hyaluronic acid.Enzyme-linked immunosorbent assay was used to measure fibronectin expression.Results：Compared with the blank control group,angiotensin Ⅱ induced cardiac fibroblast proliferation,stimulated DNA and collagen syntheses（P0.01）,and increased fibronectin,hyaluronic acid and procollagen Ⅲ concentrations（P0.01）.Xuefu Zhuyu decoction-medicated serum inhibited CF proliferation and DNA and collagen syntheses（P0.05）,and decreased the concentrations of fibroectin,hyaluronic acid and procollagen Ⅲ（P0.01）.Conclusion：Angiotensin Ⅱ can induce cardiac fibrosis,stimulate cardiac fibroblast proliferation and extracellular matrix synthesis,while Xuefu Zhuyu Decoction can prevent cardiac fibrosis,the mechanism of which may be related to inhibiting CF proliferation and extracellular matrix synthesis.
出处
《中西医结合学报》
CAS
2011年第3期313-319,共7页
Journal of Chinese Integrative Medicine
基金
国家自然科学基金资助项目(No.30472206)
上海市教育委员会资助项目(No.08cz030)
关键词
心肌纤维化
心肌重塑
中草药
血管紧张素Ⅱ
细胞外基质
cardiac fibrosis
cardiac remodeling
drugs
Chinese herbal
angiotensin Ⅱ
extracellular matrix
