香烟烟雾提取物对小鼠C2C12细胞myostatin及炎症介质的影响

　　[摘要]目的 探討香烟烟雾提取物（CSE）对小鼠C2C12细胞肌肉生长抑制素（myostatin）及炎症介质的影响。方法 不同浓度CSE和（或）myostatin-siRNA刺激和（或）干扰小鼠C2C12细胞后利用MTT检测CSE对细胞生长的影响，根据处理方式不同，实验分为4个组，分别为对照组、5% CSE组、myostatin-siRNA组、5% CSE+myostatin-siRNA组，采用Real-time PCR检测myostatin mRNA表达情况，采用Western blot检测myostatin蛋白表达情况，采用ELISA检测细胞培养液中肿瘤坏死因子-α（TNF-α）和白细胞介素8（IL-8）含量。结果 5%以上的CSE处理能显著抑制小鼠C2C12细胞增殖（P<0.05）；5%的CSE处理小鼠C2C12细胞48 h后，myostatin mRNA和蛋白表达明显高于对照组（P<0.05），细胞培养液中IL-8和TNF-α含量明显高于对照组（P<0.05）；siRNA干扰myostatin后再采用CSE处理小鼠C2C12细胞，细胞培养液中IL-8和TNF-α含量明显低于CSE处理组（P<0.05）。结论 CSE上调小鼠C2C12细胞myostatin表达，增加炎症介质释放。
　　[关键词]慢性阻塞性肺疾病；香烟烟雾提取物；肌肉生长抑制素；炎症介质
　　[中图分类号] R332 [文献标识码] A [文章编号] 1674-4721（2018）2（b）-0004-04
　　Influence of cigarette smoke extract on myostatin and inflammatory mediators in mouse C2C12 cells
　　LI Yu-ji ZHOU Xu PEI Xing-hua HUANG Li LEI Yan-jun ZHANG Lu-lu
　　The First Department of Critical Care Medicine，Peopel′s Hospital of Hu′nan Province，Changsha 410005，China
　　[Abstract]Objective To investigate the effect of cigarette smoke extract on myostatin and inflammatory mediators in mouse C2C12 cells.Methods Different concentrations of CSE and/or myostatin-siRNA stimulated or interference mouse C2C12 cells，and then detected influence of CSE on cell growth by MTT，according to the different treatment methods，the experimental research objects were divided into the control group，5% CSE group，the myostatin-siRNA group，5% CSE+myostatin-siRNA group.Real-time PCR was used to detect the expression of myostatin mRNA，Western blot was used to detect expression of myostatin protein，ELISA was used to detected tumor necrosis factor alpha （TNF-alpha） and interleukin 8 （IL-8） level in cell culture medium.Results The CSE at the concentration above 5% could significantly inhibit the proliferation of C2C12 cells in mice （P<0.05）.The expression of myostatin mRNA and protein in C2C12 cells treated with 5% CSE was significantly higher than that of the control group after 48 h （P<0.05）.The content of IL-8 and TNF-alpha in the cell culture fluid was significantly higher than that in the control group （P<0.05）.After siRNA interfered with myostatin，the mouse C2C12 cells were treated with CSE，the content of IL-8 and TNF-alpha in cell culture solution was significantly lower than that in CSE treatment group （P<0.05）.Conclusion CSE can up-regulate the expression of myostatin and increase inflammatory mediators releasing from C2C12 cells.
　　[Key words]Chronic obstructive pulmonary disease；Cigarette smoke extract；Myostatin；Inflammatory mediator