| 引用本文:蒋锋利，黄茂，蔡松，任士杰，孙文丽，张晶，赵明镜，张立山.大黄虫丸对转化生长因子-β1诱导A549细胞上皮-间充质转化的影响[J].世界中医药,2021,(24):. |
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| 大黄虫丸对转化生长因子-β1诱导A549细胞上皮-间充质转化的影响 |
| Effects of Da Huang Zhe Chong Pills on the Mesenchymal Transformation of A549 Epithelial Cells Induced by Transforming Growth Factor-β1 |
| 投稿时间：2020-06-16 |
| DOI：10.3969/j.issn.1673-7202.2021.24.007 |
| 中文关键词: 大黄虫丸 肺络干血 特发性肺纤维化 含药血清 上皮细胞-间充质转化 转化生长因子-β1 α-平滑肌肌动蛋白 Ⅲ型胶原蛋白 |
| English Keywords:Dahuang Zhechong Pills Pulmonary collateral hard blood IPF Drug-containing serum EMT TGF-β1 α-SMA Collagen Ⅲ |
| 基金项目:国家自然科学基金面上项目（81673904） |
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| 中文摘要: |
| 目的：基于“IPF患者存在肺络干血”假说,观察大黄虫丸含药血清对TGF-β1诱导后A549细胞上皮-间充质转化(EMT)及Smad3,Collagen Ⅲ表达的影响,探讨其防治特发性肺纤维化(IPF)的作用机制。方法：正常培养并取对数生长期A549细胞,TGF-β1对其诱导48 h,电子显微镜下观察A549细胞的形态学变化;选择吡非尼酮（PFD）为阳性对照药,RT-PCR法检测EMT相关基因及Smad3 mRNA的表达;Western Blottingting法检测Smad3,p-Smad3,Collagen Ⅲ蛋白表达。结果：成功获取发生EMT的A549细胞;以α-SMA mRNA表达情况判断EMT程度由高到低依次为:模型组(M),大黄虫丸含药血清大剂量组(D),西药组(P),大黄虫丸含药血清中剂量组(Z),大黄虫丸含药血清小剂量组(X),空白对照组(K);与M组比较,Z组,X组,P组中Collagen Ⅲ表达不同程度降低(P<0.05),且其在Z组与K组中表达差异无统计学意义(P>0.05);与M组及K组比较,各观察组Smad3表达差异无统计学意义(P>0.05),但对其基因及磷酸化,即Smad3 mRNA及p-Smad3的表达有影响,且与M组比较,Z组,X组,P组中均不同程度降低(P<0.05),D组中表达虽低于M组,但差异无统计学意义(P>0.05)。结论：大黄虫丸对TGF-β1诱导后A549细胞发生EMT程度与Smad3 mRNA及Collagen Ⅲ的表达有影响,推测大黄虫丸对IPF的保护机制可能与不同程度阻断或逆转了TGF-β1/Smad3介导的EMT有关。 |
| English Summary: |
| To observe the effects of Dahuang Zhechong Pills-containing serum on EMT and the expression of Smad3 and Collagen Ⅲ Protein in A549 cells after TGF-β1 induction based on the hypothesis of “the presence of pulmonary collateral hard blood in IPF patients”，and to explore its role in the prevention and treatment of idiopathic pulmonary fibrosis mechanism.Methods:Normally we grew and took A549 cells in logarithmic growth phase，TGF-β1 intervened for 48 h，observed the morphological changes of A549 cells under electron microscope; choosed pirfenidone as a positive control drug，RT-PCR method to detect the expression of EMT related genes and Smad3 mRNA; Western Blotting was used to detect the expression of Smad3,p-smad3，Collagen Ⅲ protein.Results:The A549 cells with EMT were successfully obtained; the degree of EMT was determined from α-SMA mRNA expression in order from high to low：model group(M)，large-dose group of Dahuang Zhechong Pills containing serum(D)，Western medicine group(P)，middle-dose group of Dahuang Zhechong Pills containing serum(Z)，little-dose group of Dahuang Zhechong Pills containing serum(X)，blank control group(K); Compared with M，the expression of Collagen Ⅲ in Z，X，and P decreased to varying degrees(P<0.05)，and there was no difference in the expression of Z and K(P>0.05); compared with M and K，there was no difference in the expression of Smad3 in each experimental group(P>0.05)，but it had an effect on the expression of its genes and phosphorylation，that is，the expression of Smad3 mRNA and p-smad3.All of them decreased in different degrees(P<0.05).Although the expression in D was lower than M，it was not statistically significant(P>0.05).Conclusion:Dahuang Zhechong Pills has the effect on EMT and the expression of Smad3 mRNA and Collagen Ⅲ in A549 cells after TGF-β1 induction.It is speculated that the protective mechanism of Dahuang Zhechong Pills on IPF may be related to the blocking or reversal of EMT mediated by TGF-β/Smad3 to varying degrees. |
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