Chen Ming, Lv Zhiqiang, Huang Linjie, Zhang Wei, Lin Xiaoling, Shi Jianting, Zhang Wei, Liang Ruiyun, Jiang Shanping
Department of Respiratory Medicine, Sun Yat-Sen Memorial Hospital, Institute for Respiratory disease of Sun Yat-sen University, Sun Yat-sen University, Guangzhou, Guangdong Province 510120, China.
Department of Geratology, the Second People׳s Hospital of Shenzhen, Shenzhen 518000, China.
Exp Cell Res. 2015 Feb 15;331(2):362-8. doi: 10.1016/j.yexcr.2014.10.016. Epub 2014 Oct 31.
We have reported that triptolide can inhibit airway remodeling in a murine model of asthma via TGF-β1/Smad signaling. In the present study, we aimed to investigate the effect of triptolide on airway smooth muscle cells (ASMCs) proliferation and the possible mechanism.
Rat airway smooth muscle cells were cultured and made synchronized, then pretreated with different concentration of triptolide before stimulated by TGF-β1. Cell proliferation was evaluated by MTT assay. Flow cytometry was used to study the influence of triptolide on cell cycle and apoptosis. Signal proteins (Smad2, Smad3 and Smad7) were detected by western blotting analysis.
Triptolide significantly inhibited TGF-β1-induced ASMC proliferation (P<0.05). The cell cycle was blocked at G1/S-interphase by triptolide dose dependently. No pro-apoptotic effects were detected under the concentration of triptolide we used. Western blotting analysis showed TGF-β1 induced Smad2 and Smad3 phosphorylation was inhibited by triptolide pretreatment, and the level of Smad7 was increased by triptolide pretreatment.
Triptolide may function as an inhibitor of asthma airway remodeling by suppressing ASMCs proliferation via negative regulation of Smad signaling pathway.
我们曾报道雷公藤甲素可通过TGF-β1/Smad信号通路抑制哮喘小鼠模型中的气道重塑。在本研究中，我们旨在探究雷公藤甲素对气道平滑肌细胞（ASMCs）增殖的影响及其可能机制。
培养大鼠气道平滑肌细胞并使其同步化，然后在TGF-β1刺激前用不同浓度的雷公藤甲素进行预处理。通过MTT法评估细胞增殖。采用流式细胞术研究雷公藤甲素对细胞周期和凋亡的影响。通过蛋白质免疫印迹分析检测信号蛋白（Smad2、Smad3和Smad7）。
雷公藤甲素显著抑制TGF-β1诱导的ASMC增殖（P<0.05）。雷公藤甲素剂量依赖性地将细胞周期阻滞在G1/S期。在我们使用的雷公藤甲素浓度下未检测到促凋亡作用。蛋白质免疫印迹分析显示，雷公藤甲素预处理可抑制TGF-β1诱导的Smad2和Smad3磷酸化，且雷公藤甲素预处理可提高Smad7水平。
雷公藤甲素可能通过负向调节Smad信号通路抑制ASMCs增殖，从而作为哮喘气道重塑的抑制剂发挥作用。
