miR-146a介导的白茶提取物调控Ⅱ型肺泡细胞上皮-间质转化抑制干燥综合征肺纤维化的分子机制OA
Molecular Mechanism of White Tea Extract Regulating EMT in Type Ⅱ Alveolar Cells via miR-146a to Inhibit Pulmonary Fibrosis in Sjögren's Syndrome
探究白茶提取物对干燥综合征伴肺纤维化病变Ⅱ型肺泡细胞及 microRNA-146a(miR-146a)表达水平的作用机制.采用人肺泡上皮细胞 A549 进行试验,分别设置未经转化生长因子-β1(Transforming growth factor-β1,TGF-β1)处理的空白组和经 TGF-β1 处理的 TGF-β1 诱导组(未经干预)、白茶提取物组(50 μg·mL-1 白茶提取物干预)、miR-146a mimic 组(转染 6 μL 的 miR-146a mimic 干预)、白茶提取物+miR-146a mimic 组(50 μg·mL-1白茶提取物+6 μL miR-146a mimic 干预).通过显微镜观察细胞形态,CCK-8 检测细胞活性,联合免疫荧光、蛋白印迹与实时荧光定量 PCR,检测相关蛋白及靶基因、微小 RNA 表达变化.结果显示:与空白组相比,TGF-β1诱导组的细胞活性、α 平滑肌肌动蛋白(α-SMA)荧光强度、肺表面活性物质相关蛋白 C(SP-C)、血小板衍生生长因子受体 α(PDGFRα)及 TGF-β1 表达显著升高,E-钙黏蛋白(E-cadherin)荧光强度及 miR-146a 表达显著降低(P＜0.05);与 TGF-β1 诱导组相比,白茶提取物组和 miR-146a mimic 组的细胞活性、α-SMA 荧光强度、SP-C、PDGFRα 及 TGF-β1 表达显著降低,E-cadherin 荧光强度及 miR-146a 表达显著升高(P＜0.05),但两组间无显著差异(P＞0.05);与 miR-146a mimic 组相比,白茶提取物+miR-146a mimic 组的上述指标变化更加显著.研究表明,白茶提取物可显著抑制 TGF-β1 诱导的 A549 细胞异常活化,逆转其上皮-间质转化(Epithelial-mesenchymal transition,EMT)进程并下调成纤维细胞相关活性指标,其作用机制可能与上调miR-146a 表达、调控 TGF-β1/Smad2 信号通路相关.
This paper aimed to investigate the mechanism of white tea extract on type Ⅱ alveolar cells and the expression level of microRNA-146a(miR-146a)in Sjogren's syndrome complicated with pulmonary fibrosis.Human alveolar epithelial A549 cells were divided into different groups:the blank group was not treated with transforming growth factor-β1(TGF-β1),the TGF-β1-induced group was treated with TGF-β1 without any intervention,the white tea extract group was treated with 50 μg·mL-1 white tea extract after TGF-β1 induction,the miR-146a mimic group was transfected with 6 μL miR-146a mimic after TGF-β1 induction;and the white tea extract+miR-146a mimic group was intervened with 50 μg·mL-1 white tea extract combined with 6 μL miR-146a mimic after TGF-β1 induction.Cell morphology was observed under a microscope and cell viability was detected by CCK-8 assay.The fluorescence intensities of E-cadherin and α-SMA proteins were measured by immunofluorescence.The expressions of SP-C and PDGFRα were detected by Western blotting.The expression levels of miR-146a,TGF-β1 and Smad2 were determined by qRT-PCR.The results show that compared with the blank group,the TGF-β1-induced group exhibited increased cell viability,fluorescence intensity of α-SMA protein,increased expressions of SP-C,PDGFRα and TGF-β1,as well as decreased expressions of E-cadherin and miR-146a(P<0.05).Compared with the TGF-β1-induced group,the white tea extract group and the miR-146a mimic group showed decreased cell viability,fluorescence intensity of α-SMA,expressions of SP-C,PDGFRα and TGF-β1,together with increased expressions of E-cadherin and miR-146a(P<0.05),with no statistically significant difference between the two groups(P>0.05).Compared with the miR-146a mimic group,the aforementioned indicators in the white tea extract+miR-146a mimic group changed more significantly(P<0.05).This study indicates that white tea extract can markedly inhibit the abnormal activation of A549 cells induced by TGF-β1,reverse the process of epithelial-mesenchymal transition(EMT)and downregulate fibroblast-related activity indicators.Its mechanism may be related to the up-regulation of miR-146a expression and the regulation of the TGF-β1/Smad2 signaling pathway.
张晶晶;朱跃兰;王超
中国人民武装警察部队特色医学中心,天津 300000中国人民武装警察部队特色医学中心,天津 300000中国人民武装警察部队特色医学中心,天津 300000
农业科技
干燥综合征伴肺纤维化病变白茶提取物Ⅱ型肺泡细胞转化生长因子-β1
Sjögren's syndrome with pulmonary fibrosiswhite tea extracttype Ⅱ alveolar cellsTGF-β1
《茶叶科学》 2026 (3)
535-544,10
国家自然科学基金(81874438)
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