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,舯和TransweU法观察小RNA干扰(siRNA)技术沉默Smad2及大剂量[750rag/(kg·d)]、小剂量[150Ⅱ∥(kg·d)]QC含药血清对转化生长因子Bl(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.