Results
Title

TGF-β1调控人肺泡上皮细胞中DNA甲基化酶 和端粒酶的机制.

Authors

孙梓越; 钱力; 李丹; 朱瑞芳; 韩永康; 刘学军

Abstract

Objective: Changes in DNA methylase and telomerase were observed by TGF - β1 - induced human alveolar epithelial cells A549. Methods: A549 cells were induced by TGF-β1 to establish an in vitro pulmonary fibrosis model, which was divided into a control group, TGF -β1 2μmol/L intervention group, TGF -β1 5 μmol/L intervention group, and TGF -β1 10 μmol/L intervention group. β -gal staining to detect the degree of cell senescence;ELISA and RT-qPCR detected the expression of inflammatory factors IL-1β and IL-6. RT-qPCR detects the expression of Type I collagen（COL-I）, Fibronectin 1（FN1）, Tensin-C, DNMT3a, DNMT3b, CDH1, and TERT mRNA. Western Blot method detected TERT protein expression. Results: There were almost no senescent cells in the control group, and the degree of senescence of cells in the intervention group increased with the increase of TGF - β1 concentration;IL - 1β and IL6 expression were significantly increased in the TGF - β1 intervention group（P<0. 05）;The A549 cell morphology of the control group showed a typical epithelial cell-like morphology, and the TGF-β1 intervention group had a typical mesenchymal cell-like morphology, and the Masson staining results showed that the control group cells hardly saw blue collagen, while the expression of indigen-stained collagen in the TGF-β1 intervention group was significantly increased;Compared with the control group, the expression of COL-I, FN1, Tensin-C, DNMT3a, and DNMT3b mRNA was significantly increased（P<0. 05）, the expression of mRNA of TERT and CDH1 genes was reduced（P<0. 05）, and the expression of TERT protein was further verified by Western Blot（P<0. 05）. Conclusion: TGF - β1 can induce senescence of A549 cells, and promote the high expression of inflammatory factors and fibrogenic factors, resulting in fibrosis changes. TGF-β1 promoted the expression of DNA methylase and inhibited the expression of TERT.

Subjects

TRANSFORMING growth factors-beta; IN vitro studies; REVERSE transcriptase polymerase chain reaction; INTERLEUKINS; COLLAGEN; FIBRONECTINS; IDIOPATHIC pulmonary fibrosis; STAINS & staining (Microscopy); TELOMERASE; WESTERN immunoblotting; DNA methylation; CELLULAR aging; COMPARATIVE studies; GENE expression; ENZYME-linked immunosorbent assay; PHOSPHOPROTEINS; MESSENGER RNA; EPITHELIAL cells; METABOLISM

Publication

Chinese Nursing Research, 2023, Vol 37, Issue 12, p2138

ISSN

1009-6493

Publication type

Academic Journal

DOI

10.12102/j.issn.1009-6493.2023.12.010