黄莺 徐芳 袁文胜

摘要 目的：觀察肺纤维化动物模型中人工冬虫夏草（Ccs）是否通过TGFbeta1-Smad途径来影响肺间质纤维化上皮-间质转化。方法：将50只SD大鼠随机分为空白对照组、博来霉素组（BLM组）、博来霉素+冬虫夏草组（Ccs组）、空白质粒组、siRNA组（Smad3siRNA+博来霉素+冬虫夏草），采用气管内注入博来霉素（5 mg/kg）复制大鼠肺纤维化模型，对照组给予生理盐水1.25 mL/kg。Ccs组于造模当日开始灌胃人工冬虫夏草溶液5 g/（kg·d），空白质粒组于造模第二天进行空白质粒尾静脉快速注射（3 mL/kg）。siRNA组在每天Ccs灌胃基础上于造模次日尾静脉注射Smad3 siRNA质粒（3 mL/kg）。于第28天处死动物。行HE染色，并用RT-PCR检测肺组织匀浆中上皮细胞标志物E-cad、间皮细胞标志物α-SMA以及信号通路相关分子Smad3、转化生长因子（TGF）-β1、结缔组织生长因子（CTGF）的mRNA的蛋白表达变化。结果：BLM组纤维化程度较重，Ccs组炎症程度明显减轻，纤维组织大量减少，siRNA组同Ccs组比较肺泡炎及纤维化程度有所加重;RT-PCR结果显示BLM组TGF-β1、α-SMA表达较对照组明显增高，siRNA组的α-SMA、CTGF表达较Ccs组有显著上升。结论：Ccs可能通过阻断TGFbeta1-Smad信号通路抑制肺纤维化的发生。

关键词 成纤维细胞;表型转化;TGFbeta1-Smad信号通路;人工冬虫夏草，肺间质纤维化;上皮-间质转化;转化生长因子;博来霉素

Abstract Objective：To observe the effects of cultured cordyceps sinensis（cultured cordyceps Sinensis，Ccs） in the animal model of pulmonary fibrosis induced by bleomycin（BLM） on pulmonary interstitial fibrosis by TGFbeta1-Smad pathway，and explore the mechanism of treating pulmonary fibrosis.Methods：A total of 50 rats were randomly divided into a blank control Group 10，a bleomycin Group（BLM） 10，a bleomycin and Cordyceps sinensis Group（Ccs） 10，a blank plasmid Group 10，Smad3 SiRNA gene silencing rat+bleomycin mildew element+cordyceps sinensis Group（Smad3 siRNA+BLM+CCS） 10.tracheotomy injected bleomycin（5 mg） was used to replicate mouse pulmonary fibrosis model，the control group was given saline 1.25 mL/kg.Ccs Group on the day of the modelling began to stomach artificial cordyceps sinensis solution 5 g/（kg·d），the blank plasmid group in the second day of the model for the rapid injection of empty particles of the tail vein（3 mL/kg）.The Smad3 siRNA+BLM+CCS group was injected into the Smad3 siRNA plasmid（3 mL/kg） at the end of the day after the injection of CCS on a daily basis.The animals were executed on the 28th day.In the left lung tissue，hematoxylin-eosin（HE） staining was used to observe the degree of alveolar inflammation and pulmonary fibrosis，and the right lung was detected by RT-PCR to detect the E-cad of the upper and middle skin cells of lung homogenate，the α-SMA intercellular markers and the signaling pathway related molecular Smad3，TGF-β1，CTGF Changes in the expression of mRNA protein.Results：In BLM Group，the degree of fibrosis was heavy，the degree of inflammation in the CCS group decreased markedly，the fibrous tissue was greatly reduced，and the degree of alveolar inflammation and fibrosis in Smad3 SiRNA+BLM+Ccs group was worse than that in CCS group.The results of RT-PCR showed that TGF-β1 and α-SMA expression in BLM group increased significantly The expression of α-SMA，and CTGF in SiRNA+BLM+CCS group increased significantly compared with that in CCS group.Conclusion：CCS may inhibit the occurrence of pulmonary fibrosis by blocking the TGFbeta1-Smad signaling pathway and reduce the effect of pulmonary fibrosis by inhibiting the activation of the TGFbeta1-Smad signaling pathway.

Keywords Fibroblast; Transit; TGFbeta1-Smad signaling pathway; Cultured cordyceps sinensis; Pulmonary fibrosis; Epithelial-interstitial transformation; Transforming growth factor; Bleomycin

中图分类号：R282文献标识码：Adoi：10.3969/j.issn.1673-7202.2021.24.008

肺纤维化（PF）是一种原因不明且持续性进展的疾病，目前尚无特效治疗药物。国内外研究表明，肺成纤维细胞的增殖、转化是其发生机制的关键。上皮-间质转化（EMT）被认为是成纤维细胞的一个重要来源[1-2]。近年来研究显示，TGFbeta1-Smad信号通路与肺纤维化发生发展关系密切[3-4]。……