强心饮对肿瘤坏死因子α诱导的大鼠心肌成纤维细胞增殖和胶原合成以及激活蛋白1 mRNA表达的影响

Activator protein-1 mRNA expression,cell proliferation and collagen synthesis induced by tumor necrosis factor-α in cardiac fibroblasts from rats and the effect of Qiangxin Decoction

目的:研究激活蛋白1(activator protein-1,AP-1)在肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)诱导新生大鼠心肌成纤维细胞(cardiac fibroblasts,CFs)增殖、胶原合成中的作用,探讨强心饮抑制心肌纤维化的作用机制。方法:胰酶消化法分离培养乳鼠CFs,建立TNFα诱导新生大鼠CFs纤维化模型。将CFs按培养方式不同分为正常对照组、模型组和强心饮5%含药血清组、10%含药血清组和20%含药血清组。治疗24h后,采用荧光实时定量聚和酶链式方法检测不同浓度强心饮对CFs AP-1 mRNA表达的影响,甲基噻唑基四唑法检测强心饮对CFs增殖的影响;羟脯氨酸法检测对CFs胶原合成量的影响。结果:TNF-α作用CFs24h后,与空白对照组相比,AP-1 mRNA表达明显增加(P<0.05),细胞增殖和胶原合成增加(P<0.05);强心饮干预后,CFs增殖和胶原合成减少(P<0.05),AP-1 mRNA表达下调(P<0.05)。结论:强心饮能够抑制TNF-α诱导的CFs增殖和胶原合成增加,具有抗心肌纤维化作用,其机制可能与下调AP-1 mRNA表达有关。

Objective： To investigate the function of activator protein-1 （AP-1） in proliferation and collagen cardiac fibroblasts （CFs） in neonatal rats induced by tumor necrosis factor-α （TNF-α）, and to mechanism of Qiangxin Decoction （QXD）, a compound traditional Chinese herbal medicine, cardiac fibrosis. Methods： CFs derived from neonatal rats were cultured with enzymatic was induced by TNF-α. The CFs were divided into normal contro synthesis of explore the in reversing dissociation, and fibrosis of the CFs group, untreated group, 5% QXD- containing serum group, 10% QXD-containing serum group and 20% QXD-containing serum group. After 24- hour culture of QXD-containing serum, AP-1 mRNA was detected by real-time fluorescent quantitative polymerase chain reaction; proliferation and collagen synthesis of CFs were assayed by thiazolyl blue assay （MTT） and measured by hydroxyproline respectively, in order to determine the effect of different dosage decoction on the proliferation and collagen synthesis of CFs. Results： After 24-hour stimulation of TNF-α in CFs, compared with those in normal control group, the expression of AP-1 mRNA and cardiac fibroblast proliferation and collagen synthesis increased significantly （P〈0.05）. However, Qiangxin Decoction could reduce the expression of AP-1 mRNA and decreased the proliferation and collagen synthesis of CFs significantly （P〈0.05）. Conclusion： Qiangxin Decoction can reverse cardiac fibrosis, which may nhibit proliferation and collagen synthesis of CFs induced by TNF-α, and be related to its down-regulation of the expression of AP-1.