| 李玥珂,李雯,唐庆.Wilms瘤 1-相关蛋白介导的 N6-腺苷甲基化修饰对转化生长因子 β诱导的肺成纤维细胞活性和纤维化的影响[J].安徽医药,2026,30(2):291-295. |
| Wilms瘤 1-相关蛋白介导的 N6-腺苷甲基化修饰对转化生长因子 β诱导的肺成纤维细胞活性和纤维化的影响 |
| Effect of Wilms' tumor 1-associated protein-mediated modification of N6-methyladenosine on the cell viability and fibrosis of transforming growth factor β-induced lung fibroblasts |
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| DOI：10.3969/j.issn.1009-6469.2026.02.015 |
| 中文关键词: 肺纤维化 N6-腺苷甲基化 Wilms瘤 1-相关蛋白 转化生长因子 β 成纤维细胞 |
| 英文关键词: Pulmonary fibrosis N6-methyladenosine Wilms' tumor 1-associated protein Transforming growth factor-β Fibro-blasts |
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| 中文摘要: |
| 目的探讨 Wilms瘤 1-相关蛋白( WTAP)介导的 N6-腺苷甲基化( m6A)修饰对转化生长因子 β(TGF-β)诱导的肺成纤维细胞活性和纤维化的影响。方法 2022年 6月至 2023年 4月将肺成纤维细胞( HFL1)采用随机数字表法分为 6组:对照组、 TGF-β组、 TGF-β+si-NC组、 TGF-β+si-WTAP组、 TGF-β+OE-NC组和 TGF-β+OE-WTAP组。荧光定量聚合酶链反应(qPCR)检测 WTAP mRNA表达水平，蛋白质印迹法检测 WTAP、α-平滑肌肌动蛋白( α-SMA)、 Ⅰ型胶原和 Ⅲ型胶原的蛋白表达；免疫荧光检测 α-SMA表达； CCK-8检测细胞活性；试剂盒检测总 m6A水平，以及 Ⅰ型胶原和 Ⅲ型胶原水平。结果与对照组相比， TGF-β处理诱导成纤维细胞中总 m6A水平升高( 0.58±0.07比 0.26±0.04)，而且 WTAP的 mRNA(3.83±0.57比 1.00±0.10)和蛋白( 2.25± 0.31比 1.00±0.11)表达水平上调( P<0.05)；抑制 WTAP表达降低 TGF-β诱导的总 m6A水平( P<0.05)，降低细胞活性( P<0.05)，抑制 TGF-β诱导的 α-SMA(P<0.05)、 Ⅰ型胶原( P<0.05)和 Ⅲ型胶原( P<0.05)蛋白表达上调，减少 TGF-β诱导的 Ⅰ型胶原( P< 0.05)和 Ⅲ型胶原( P<0.05)沉积； WTAP过表达提高 TGF-β诱导的总 m6A水平( P<0.05)增强细胞活性( P<0.05)，上调 TGF-β诱导的 α-SMA(P<0.05)、 Ⅰ型胶原( P<0.05)和 Ⅲ型胶原( P<0.05)蛋白表达，加强TGF-β诱导，的Ⅰ型胶原( P<0.05)和 Ⅲ型胶原( P<0.05)沉积。结论 TGF-β诱导的肺成纤维细胞纤维化与 WTAP介导的 m6A修饰水平升高有关。 |
| 英文摘要: |
| Objective To investigate the effect of Wilms' tumor 1-associated protein (WTAP)-mediated modification of N6-methyl-adenosine (m6A) on the cell viability and fibrosis of transforming growth factor β (TGF-β)-induced lung fibroblasts.Methods Lung fi-broblasts HFL1 were randomly divided into 6 groups: control group, TGF-β group, TGF-β+si-NC group, TGF-β+si-WTAP group, TGF-β+OE-NC group and TGF-β+OE-WTAP group from June 2022 to April 2023. Quantitative polymerase chain reaction (qPCR) was usedto detect the mRNA expression of WTAP. Western blot was used to detect the protein expression of WTAP, α-smooth muscle actin (α -SMA), Collagen Ⅰ and Collagen Ⅲ. The expression of α-SMA was detected by immunofluorescence. Cell counting kit-8 was used to detect cell viability. Total m6A level, and the levels of Collagen I and Collagen Ⅲ were determined by commercial kits.Results Com-pared with the control group, TGF-β significantly increased the total m6A level (0.58±0.07 vs. 0.26±0.04) (P<0.05), and significantly up-regulated the mRNA (3.83±0.57 vs. 1.00±0.10) and protein expression levels (2.25±0.31 vs. 1.00±0.11) of WTAP (P<0.05). WTAP inhibition decreased the total m6A level induced by TGF-β(P<0.05), decreased cell viability (P<0.05), inhibited TGF-β-induced up-regulation of α-SMA (P<0.05), Collagen Ⅰ (P<0.05) and Collagen Ⅲ (P<0.05) protein, and reduced TGF-β-induced deposition of Col-lagen Ⅰ(P<0.05) and Collagen Ⅲ(P<0.05). WTAP overexpression elevated TGF-β-induced total m6A level (P<0.05), enhanced cell vi-ability (P<0.05), up-regulated TGF-β-induced α-SMA (P<0.05), Collagen Ⅰ (P<0.05) and Collagen Ⅲ (P<0.05) protein expression, and increased TGF-β-induced deposition of Collagen Ⅰ (P<0.05) and Collagen Ⅲ (P<0.05).Conclusion The lung fibroblast fibrosis induced by TGF-β is associated with increased WTAP-mediated m6A modification. |
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