MiR-23b通过调节TGF-β1/SMAD3信号通路对博莱霉素所致大鼠肺纤维化及自噬水平的影响
| 作者: |
1谢健龙,
2区泳芳,
2陈永华,
3常岸芷,
3,4缪辉来
1 广东医科大学附属医院心胸外科，广东 湛江 524001 2 广东医科大学附属医院病理诊断与研究中心，广东 湛江 524001 3 广东医科大学附属第二医院肝胆外科，广东 湛江 524003 4 东莞寮步医院普外科，广东 东莞 523400 |
| 通讯: |
缪辉来
Email: Miaohl-gdwk@gdmu.edu.cn |
| DOI: | 10.3978/j.issn.2095-6959.2022.06.001 |
| 基金: | 国家自然科学基金(82070637)；东莞市社会科技发展重点项目(2018507150391621)；湛江市非资助科技攻关计划项目(2021B01454)；广东医科大学青年培育基金项目(GDMUQ2021010)。 |
摘要
Effect of miR-23b on the level of pulmonary fibrosis and autophagy induced by bleomycin by regulating TGF-β1/SMAD3 signaling pathway
CorrespondingAuthor: MIAO Huilai Email: Miaohl-gdwk@gdmu.edu.cn
DOI: 10.3978/j.issn.2095-6959.2022.06.001
Foundation: This work was supported by the National Natural Science Foundation (82070637), the Social Science and Technology Development Key Project of Dongguan (2018507150391621), the Non-funded Science and Technology Research Project of Zhanjiang (2021B01454), and the Guangdong Medical University Youth Cultivation Fund Project (GDMUQ2021010), China.
Abstract
Objective: To explore the effects of miR-23b on bleomycin-induced pulmonary fibrosis (PF) and the effects of autophagy in rats by regulating the transforming growth factor-β1(TGF-β1)/mothers against decapentaplegic homolog 3 (SMAD3) signaling pathway. Methods: The rat PF model was induced by bleomycin (BLM) and divided into a control group, a PF group, an agomiR-negative control (NC) group, and an agomiR-23b group. Bioinformatics prediction and dual luciferase experiments were used to verify the regulation of miR-23b on SMAD3; hematoxylin-eosin stain (HE) and Masson staining were conducted to observe the pathological changes of rat lung tissue; immunohistochemical staining was conducted to observe the expression levels of type I collagen (collagen-I), α-smooth muscle actin (α-SMA), yeast autophagy related gene 6 mammalian homolog (beclin-1), and light chain 3-II (LC3-II) in rat lung tissue; real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) was used to detect mRNA expression of miR-23b and SMAD3 in lung tissue; Western blotting was used to detect the protein expression of TGF-β1, SMAD3, and SMAD7 in lung tissues. HFL1 cells were divided into a control group, a PF group, an agomiR-NC group, and an agomiR-23b group; RT-qPCR was used to detect the expression of miR-23b and SMAD3 mRNA in each group of cells; Transwell chamber experiment was conducted to detect cell migration and invasion ability of each group; transmission electron microscope was applied to observe the formation of autophagic vesicles of each group of cells. Results: Compared with the control group, the degree of PF in the PF group and the agomiR-NC group was more severe; the expression levels of SMAD3 mRNA and protein of collagen-I, α-SMA, TGF-β1, and SMAD3 were significantly increased, while expression levels of miR-23b and protein of beclin-1, LC3-II, and SMAD7 were significantly reduced in the lung tissues; the expression level of SMAD3 mRNA and the migration and invasion ability were increased, and the autophagy activity was reduced in HFL1 cells (all P<0.05). Compared with the PF group, the degree of PF in the agomiR-23b group was effectively improved; the expression levels of protein of collagen-I, α-SMA, TGF-β1, and SMAD3 were significantly reduced, while the expression levels of beclin-1, LC3-II, and SMAD7 protein were significantly increased in the lung tissue; the expression level of SMAD3 mRNA and the migration and invasion ability were reduced, and the autophagy activity was increased in HFL1 cells (all P<0.05). Conclusion: MiR-23b inhibits the expression of SMAD3, inhibits the migration of lung fibroblasts, reduces the degree of lung fibrosis, and improves autophagy active. Its mechanism may be related to the regulation of the TGF-β1/SMAD3 pathway.