The Journal of Practical Medicine >
Effects of overexpression of HSG gene on Wnt signaling pathway and MAPK signaling pathway in COPD rat airway fibroblasts
Received date: 2023-06-12
Online published: 2023-12-11
Objective To study the effect of overexpression of HSG gene (MFN2) on Wnt signaling pathway in airway fibroblasts of rats with chronic obstructive pulmonary disease (COPD). Methods The rat COPD model was induced by protease, and then the airway fibroblasts of COPD model rats were isolated in primary culture. The expression vector of HSG gene was constructed and transfected into rat airway fibroblasts to obtain rat airway fibroblasts overexpressing HSG. In Experiment 1, cells were treated with phorbol ester (TPA), a Wnt signaling pathway agonist, and the expressions of HSG and Wnt5a were detected by WB and qPCR; In Experiment 2, the cells were treated with EGF factor, an agonist of MAPK signal pathway, and the expressions of HSG and ERK1/2 were detected by WB and qPCR; Moreover, ELISA was used to detect TGF in cell culture medium in each experiment-β1. The contents of PDGF and MMP-9, CCK8, cell proliferation and apoptosis were detected by flow cytometry. Results Airway fibroblasts were successfully isolated from COPD model rats. In Experiment 1 and Experiment 2, WB and qPCR showed that overexpression of HSG could inhibit the expression of genes Wnt5a and ERK1/2. After treatment with corresponding agonists, the expression of these genes was up-regulated and the expression of HSG was inhibited; The results of ELISA showed that overexpression of HSG could reduce TGF-β1. The contents of PDGF and MMP-9, TGF after treatment with corresponding agonists-β1. The contents of PDGF and MMP-9 increased, but overexpression of HSG still had inhibitory effect. CCK8 detection showed that overexpression of HSG could inhibit cell proliferation, while flow cytometry showed that overexpression of HSG could significantly increase the apoptosis rate, and the corresponding agonist treatment would reduce the effect of overexpression of HSG on cell apoptosis. Flow cytometry showed that the effect of overexpression of HSG on cell cycle was not obvious. Conclusion Overexpression of HSG gene in airway fibroblasts of COPD rats can inhibit Wnt, MAPK and other signal pathways, and overexpression of HSG gene can promote fibroblast apoptosis and inhibit cell proliferation. It can be speculated that promoting HSG expression can inhibit airway fibrosis of COPD rats.
Yi YANG , Xun ZHOU , Jun ZHANG , Bo LI , Xinxing WANG , Xian LUO , Zhengxing. GE . Effects of overexpression of HSG gene on Wnt signaling pathway and MAPK signaling pathway in COPD rat airway fibroblasts[J]. The Journal of Practical Medicine, 2023 , 39(22) : 2921 -2927 . DOI: 10.3969/j.issn.1006-5725.2023.22.011
| 1 | ROCKER G M, AMAR C, LAFRAMBOISE W L, et al. Spreading improvements for advanced COPD care through a Canadian Collaborative[J]. Int J Chron Obstruct Pulmon Dis, 2017, 12: 2157-2164. |
| 2 | WU X J, JIA B L, LUO X B, et al. Glucocorticoid Alleviates Mechanical Stress-Induced Airway Inflammation and Remodeling in COPD via Transient Receptor Potential Canonical 1 Channel[J].Int J Chron Obstruct Pulmon Dis, 2023, 18: 1837-1851. |
| 3 | SU Y C, HAN W H, KOVACS-K A, et al. HDAC6 Activates ERK in Airway and Pulmonary Vascular Remodeling of Chronic Obstructive Pulmonary Disease[J].Am J Respir Cell Mol Biol, 2021, 65: 603-614. |
| 4 | MAHMOOD M Q, REID D, WARD C, et al. Transforming growth factor (TGF) β1 and Smad signalling pathways: A likely key to EMT-associated COPD pathogenesis[J]. Respirology, 2017, 22(1):133. |
| 5 | LOU Y, RONG L I, LIU J, et al. Construction and efficacy identification of the lentiviral vector harboring RNAi based on hyperplasia suppressor gene(HSG)[J]. Beijing Biomedical Engineering, 2015. |
| 6 | 罗卫. Mfn2/HSG与恶性肿瘤[J]. 实用肿瘤学杂志, 2013, 27(5):3. |
| 7 | ROGERS S, ZHANG C T, ANAGNOSTIDIS V, et al. Cancer-associated fibroblasts influence Wnt/PCP signaling in gastric cancer cells by cytoneme-based dissemination of ROR2[J] .Proc Natl Acad Sci U S A, 2023, 120: e2217612120. |
| 8 | 田晨,刘志辉, 孟繁荣,等. 香烟烟雾提取物诱导16HBE细胞MUC5AC上调的机制 [J]. 实用医学杂志, 2022, 38(18): 2309-2317. |
| 9 | CHILOSI M, POLETTI V, ZAMò A, et al. Aberrant Wnt/beta-catenin pathway activation in idiopathic pulmonary fibrosis.[J]. Am J Pathol, 2003, 162(5):1495. |
| 10 | K?NIGSHOFF M, BALSARA N, PFAFF E M, et al. Functional Wnt Signaling Is Increased in Idiopathic Pulmonary Fibrosis[J]. PLos One, 2008, 3(5):e2142. |
| 11 | PENG Y, ZHANG X, MA Q, et al. MiRNA-194 activates the Wnt/β-catenin signaling pathway in gastric cancer by targeting the negative Wnt regulator, SUFU[J]. Cancer Letters, 2016, 385:117-127. |
| 12 | WANG R, SUN Q, WANG P, et al. Notch and Wnt/β-catenin signaling pathway play important roles in activating liver cancer stem cells[J]. Oncotarget, 2016, 7(5):5754-5768. |
| 13 | SU Z J, SONG J X, WANG Z Y, et al.Tumor promoter TPA activates Wnt/β-catenin signaling in a casein kinase 1-dependent manner[J]. PNAS, 2018,115 (32): E7522-E7531. |
| 14 | MASCKAUCHáN T N, AGALLIU D, VORONTCHIKHINA M, et al. Wnt5a signaling induces proliferation and survival of endothelial cells in vitro and expression of MMP-1 and Tie-2[J]. Mol Biol Cell,2006,17:5163-5172. |
| 15 | WANG J, HUANG J N, WANG L L, et al. Urban particulate matter triggers lung inflammation via the ROS-MAPK-NF-κB signaling pathway[J]. J Thorac Dis, 2017, 9(11): 4398-4412. |
| 16 | CICCHINI M, BUZA E L, SAGAL K M, et al. Context dependent effects of amplified MAPK signaling during lung adenocarcinoma initiation and progression[J].Cell Rep, 2017,18(8): 1958-1969. |
| 17 | CHRISTMAN J W, YE R D, GANG W, et al. Pivotal Role of Mitogen-Activated Protein Kinase-Activated Protein Kinase 2 in Inflammatory Pulmonary Diseases[J].Curr Protein Pept Sci, 2016, 17(4): 332-342. |
| 18 | SHEN B, SHI J P, ZHU Z X, et al. EGFR inhibition overcomes resistance to FGFR4 inhibition and potentiates FGFR4 inhibitor therapy in hepatocellular carcinoma [J]. Mol Cancer Ther, 2023. PMID: . |
| 19 | ?ERVENKA I, WOLF J, MA?EK J, et al. Mitogen-activated protein kinases promote WNT/beta-catenin signaling via phosphorylation of LRP6[J]. Mol Cell Biol, 2011,31:179-189. |
| 20 | 姚兰, 李均. 肾纤维化MAPK相关信号通路及中医药干预的研究进展[J]. 中华中医药杂志, 2015, 30(7):3. |
| 21 | WU X, TU X, JOENG K S, et al. Rac1 activation controls nuclear localization of beta-catenin during canonical Wnt signaling[J]. Cell, 2008,133(2):340-353. |
| 22 | 张婵娟, 覃丽. Wnt信号通路介导胆固醇逆向转运与炎症应答的交互作用[J]. 中国动脉硬化杂志, 2018, 26(8):8. |
| 23 | WESTERGREN‐THORSSON G, BAGHER M, ANDERSSON‐SJ?LAND A, et al. VEGF synthesis is induced by prostacyclin and TGF‐β in distal lung fibroblasts from COPD patients and control subjects: Implications for pulmonary vascular remodelling[J]. Respirology, 2018, 23(1): 68-75. |
| 24 | 张喆,李娜,刘谦,等. 阿胶对COPD模型小鼠的保护作用以及对MMP-2、MMP-9、TGF-β1水平的影响[J]. 基因组学与应用生物学,2018,37(4):1813-1819. |
| 25 | YI E K, LIN B T, ZHANG Y,et al. Smad3-mediated lncRNA HSALR1 enhances the non-classic signalling pathway of TGF-β1 in human bronchial fibroblasts by binding to HSP90AB1[J]. Clin Transl Med, 2023, 13: e1292. |
| 26 | 王玉娟,薛亚妮,陈伟,等. 痰热清注射液对COPD患者血清TGF-β与MMP-9水平的影响[J]. 现代生物医学进展,2017,17(22):4325-4329。 |
| 27 | 李育娟. 盐酸戊乙奎醚对慢性阻塞性肺疾病患者血清TNF-α和MMP-9水平影响分析[J]. 解剖学研究,2017,39(1):35-37. |
| 28 | 童卫泉,杨宏宽,夏婷婷,等. 保肺定喘汤对COPD气道重塑大鼠MMP-9、VEGF水平的影响[J]. 江西中医药大学学报,2016,28(6):67-70. |
| 29 | ZOU W, HE F, LIU S, et al. PM2.5 Induced the Expression of Fibrogenic Mediators via HMGB1-RAGE Signaling in Human Airway Epithelial Cell [J]. CAN RESPIR J, 2018, 2018(3):1-10. |
| 30 | HELDIN C H. Structural and functional studies on platelet-derived growth factor[J]. EMBO J, 1992,11 (12): 4251-4259. |
| 31 | WANG L L, LIU D, WEI J, et al. MiR-543 Inhibits the Migration and Epithelial-To-Mesenchymal Transition of TGF-β-Treated Endometrial Stromal Cells via the MAPK and Wnt/β-Catenin Signaling Pathways[J]. Pathol Oncol Res, 2021, 27: 1609761. |
| 32 | YANG S, LIU Y, LI M Y, et al. FOXP3 promotes tumor growth and metastasis by activating Wnt /β-catenin signaling pathway and EMT in non-small cell lung cancer[J]. Mol Cancer,2017,16(1): 124. |
| 33 | WOJCIECHOWSKI M C, MAHMUTOVIC L, SHU D Y, et al .ERK1 /2 signaling is required for the initiation but not progression of TGFβ-induced lens epithelial to mesenchymal transition ( EMT) [J]. Exp Eye Res,2017,159: 98-113. |
| 34 | 田晨,刘志辉,孟繁荣,等. 香烟烟雾提取物诱导16HBE细胞MUC5AC上调的机制 [J]. 实用医学杂志, 2022, 38(18): 2309-2317. |
/
| 〈 |
|
〉 |