芩丹胶囊对成纤维细胞增殖与迁移的影响

Inhibitory effect of the Qindan capsule on proliferation and migration of adventitial fibroblasts

目的探讨芩丹胶囊(QC)含药血清对血管外膜成纤维细胞(AF)增殖和迁移的影响。方法组织贴块法原代培养大鼠胸主动脉成纤维细胞,MTT和Transwell法观察小RNA干扰(siRNA)技术沉默Smad2及大剂量[750㎎/(㎏.d)]、小剂量[150㎎/(㎏.d)]QC含药血清对转化生长因子β1(TGF-β1)诱导AF增殖和迁移的影响;实时定量PCR及Western-blot检测Smad2和Smad7的基因和蛋白表达情况。结果 siRNA-Smad2及大、小剂量QC含药血清均能显著抑制AF的增殖和迁移(P<0.05);用药组Smad2基因及蛋白的表达较单纯刺激组显著降低,Smad7则显著升高(P均<0.01)。结论 QC含药血清能显著抑制AF增殖和迁移,其机制可能与Smads信号转导通路有关。

Objective To investigate effect and mechanism of Qindan capsule （QC）-containing serum on proliferation and migration of adventitial fibroblasts （AF）. Methods AFs from the thoracic aorta of rats were cultured in vitro by a tissue explant. The proliferation and migration of AF affected by siRNA-Smad2 and QC-containing serum with high [750 mg/（kg·d） ] or low doses [ 150 mg/（kg·d） ] were detected by MTT and Transwell. mRNA and protein expressions of Smad2 and Smad7 were observed by real-time PCR and Western blot, respectively. Results After stimulation by TGF-β1 for 24 h, the proliferation and migration of AF were higher compared with those untreated. Knockdown of Srnad2 and QC-containing serum with high and low doses inhibited proliferation and migration of AF （ P 〈 0.05 ）. And drug-containing serum could downregulate expression of Smad2, while up-regulating expression of Smad7 （P 〈0.01 ）. Conclusion QC-containing serum has a significant inhibitory effect on proliferation and migration of AF induced by TGF-β1. The mechanism may be related to the TGF-β1/ Smads signaling pathway.