【摘要】 目的:观察氧化苦参碱(Oxymatrine,OM)干预对转化生长因子β1 (Transforming growth factor-β1,TGF-β1)刺激的SD大鼠心肌成纤维细胞(Cardialfibroblasts,CFs)增殖、分化的抑制作用,并探讨其作用机制。方法:体外培养新生大鼠CFs,分离纯化后分为对照组(使用无血清DMEM培养)、TGF-β1组(单纯TGF-β1诱导)和观察组(使用TGF-β1加OM干预) 3组。MTT法检测CFs增殖情况,ELISA法检测Ⅰ型胶原(Collagen-Ⅰ,CoL-Ⅰ)和Ⅲ型胶原(Collagen-Ⅲ,CoL-Ⅲ)含量,RT-PCR法检测Smad2 mRNA表达,Westorn blot法检测增殖细胞核抗原(PCNA)、α-平滑肌肌动蛋白(α-SMA)、Smad2及Smad3蛋白表达情况。结果:与对照组比较,TGF-β1组CFs增殖水平、CoL-Ⅰ、CoL-Ⅲ、PCNA、α-SMA、Smad2 mRNA及Smad2/3蛋白表达均明显升高(P <0.05);与TGF-β1组比较,观察组上述指标均降低(P <0.05),除CoL-Ⅰ外,差异均有统计学意义(P <0.05)。结论:OM能够明显抑制TGF-β1诱导CFs增殖、分化水平,降低胶原蛋白的合成水平,作用机制与其能够干预Smad2及Smad3蛋白表达有关,具有临床应用价值。更多还原

【Abstract】 Objective:To observe the inhibitory effect of oxymatrine(OM) on the proliferation and differentiation of cardial fibroblasts(CFs) in SD rats induced by transforming growth factor-β1(TGF-β1) and explore its mechanism. Methods:Separated and purified CFs of neonatal rats in vitro,and divided them into the control group(cultured by serum-free DMEM),the TGF-β1 group(simply induced by TGF-β1) and the observation group(intervened by TGF-β1 and OM). Detected the proliferation of CFs by MTT method,the content of collagen-I(CoL-I) and collagen-Ⅲ(CoL-Ⅲ) by ELISA method and the expression of m RNA of Smad2 by RT-PCR method. Detected the expressions of proliferating cell nuclear antigen(PCNA),α-smooth muscle actin(α-SMA) and protein expressions of Smad2 and Smad3 by Westorn blot method. Results:Compared with those in the control group,the proliferation of CFs and the expressions of CoL-I,CoL-Ⅲ,PCNA,α-SMA and protein expressions of Smad2 Smad2/3 in the TGF-β1 group were obviously increased(P < 0.05). Compared with those in the TGF-β1 group,the above indexes(except CoL-I)in the observation group were decreased,differences were significant(P < 0.05). Conclusion:OM can obviously inhibit the levels of proliferation and differentiation of CFs induced by TGF-β1 and decrease the translation level of collagen. Its mechanism is related to the intervention of the protein expressions of Smad2 and Smad3,which maintains high value in clinical practice.更多还原