安徽医科大学学报2025，Vol.60Issue(1)：66-72,7.DOI:10.19405/j.cnki.issn1000-1492.2025.01.009
靶向沉默DNMT3A对小鼠肺成纤维细胞胶原沉积、增殖及迁移活性的影响
Effect of targeted silencing of DNMT3A on collagen deposition,proliferation and migration activity of mouse lung fibroblasts
摘要
Abstract
Objective To investigate the effect of targeted silencing of DNA methyltransferase 3A(DNMT3A)on collagen deposition,proliferation and migration activity of mouse lung fibroblasts(PFs).Methods In order to en-sure the proliferation and migration activity of primary fibroblasts,the lung tissues of neonatal C57 suckling mice were taken,PFs were extracted after being sheared,and the morphology was observed and identified under the mi-croscope.PFs cells were activated by 5 ng/ml TGF-β1 for 24 h after cell attachment,and DNMT3A silencing model was constructed by small interfering RNA;The experiment was divided into control group,TGF-β1 group,TGF-β+siRNA-NC group and TGF-β1+siRNA-DNMT3A group.The protein expressions of DNMT3A,α-smooth mus-cle actin(α-SMA)and Collagen Ⅰ were detected by Western blot;Real time quantitative reverse transcription polymerase chain reaction(RT-qPCR)was used to detect the mRNA expression changes of DNMT3A,α-SMA and Collagen Ⅰ.The proliferation ability of PFs was detected by CCK-8 and EdU staining;the migration ability of PFs was detected by scratch test and Transwell migration test.Results Compared with the control group,TGF-β1 in-duced the increase of DNMT3A in the activated PFs cell group(P＜0.01),the protein and mRNA levels of fibro-sis and proliferation related indicators α-SMA and Collagen Ⅰ also increased(all P＜0.05),and the proliferation and migration ability of PFs increased(all P＜0.000 1).Compared with the siRNA-NC group,the protein expres-sion levels of DNMT3A(P＜0.000 1)and related indicators α-SMA(P＜0.01)and Collagen Ⅰ(P＜0.01)sig-nificantly decreased in the DNMT3A silencing group by Western blot,and the mRNA levels of DNMT3A,α-SMA and Collagen Ⅰ by RT-qPCR also decreased(all P＜0.001),and the proliferation(P＜0.01)and migration abil-ity(P＜0.05)of PFs cells decreased compared with the control group.Conclusion Silencing DNMT3A can in-hibit the deposition of collagen and the proliferation of PFs.DNMT3A can promote the proliferation and migration of PFs,and then promote the activation of PFs and the development of pulmonary fibrosis.This process may be regu-lated by DNA methylation modification.关键词
DNMT3A/DNA甲基化/肺成纤维细胞/增殖/迁移/肺纤维化Key words
DNMT3A/DNA methylation/lung fibroblasts/proliferation/migration/pulmonary fibrosis分类
医药卫生引用本文复制引用
汪先晨,尤峻柏,凌辉,范家好,陈齐,陶辉,沙纪名..靶向沉默DNMT3A对小鼠肺成纤维细胞胶原沉积、增殖及迁移活性的影响[J].安徽医科大学学报,2025,60(1):66-72,7.基金项目
安徽省高校自然科学研究项目(编号:2023AH040376) Natural Science Research Project of Anhui Educational Committee(No.2023AH040376) （编号:2023AH040376）
