Glucagon like peptide 1(28-36)amide targets mitochondrial related pathways to inhibit transforming growth factor β-mediated activation of pulmonary fibroblasts
Dong Zehai
Hou Sitong
Shang Linjiao
Liu Yuxin
Lyu Xiaoxi
Yu Yang
Wang Xiaoxia
Abstract：Objective To investigate the effect of glucagon like peptide 1(GLP-1)metabolite GLP-1(28-36)amide on transforming growth factor-β(TGF-β)-mediated proliferation and activation of pulmonary fibroblasts and its possible mechanism.Methods Mouse lung fibroblasts(MLg cells)were divided into control group,TGF-βgroup,TGF-β+liraglutide group,TGF-β+GLP-1(28-36)amide group.The phosphorylation level of Smad homolog 3(SMAD3)in MLg cells was detected by Western blotting.Cell counting kit-8 was used to detect the proliferation of MLg cells.Ligand fishing of MLg cell lysates was performed by biotin-labeled GLP-1(28-36)amide[BIOT(28-36)]and streptomycin agarose.The protein samples obtained from fishing were analyzed by high performance liquid chromatography-tandem mass spectrometry.According to the mass spectrometry data,the top 100 abundant proteins were screened for bioinformatics analysis to further screen the proteins that might interact with GLP-1(28-36)amide.Autodock vina1.5.6 software was used to verify the binding between proteins.Results Compared with the control group,the phosphorylation level of SMAD3 in MLg cells was increased(P＜0.01)and the cell proliferation rate was increased(P＜0.05)in TGF-β group.Compared with TGF-β group,the phosphorylation level of SMAD3 in TGF-β+GLP-1(28-36)amide group was decreased[(0.677±0.046)vs(1.414±0.037)](P＜0.05),and there was no significant difference in proliferation rate(P＞0.05).Bioinformatics analysis showed that mitochondrial protein YME1-like 1atpase(YME1L1),phosphoglycerate mutase 5(PGAM5),mitochondrial translation elongation factor Tu(TUFM),heat shock protein family D member 1(HSPD1),solute carrier family 25 member 5(SLC25A5)may be related to GLP-1(28-36)amides bind to each other.Autodock vina1.5.6 software was used for molecular Docking and Pymol 3.1 software was used for visualization.The docking scores of the above-mentioned proteins with GLP-1(28-36)amide were all＜-4 kcal/mol.Conclusion GLP-1(28-36)amide can inhibit TGF-β-mediated activation of pulmonary fibroblasts,which may act through mitochondrial related pathways.The proteins that can directly bind to GLP-1(28-36)amide may be mitochondrial proteins such as YME1L1,PGAM5,TUFM,HSPD1,and SLC25A5.
Keywords：Glucagon like peptide 1(28-36)amidePulmonary fibrosisTransforming growth factor-βMitochondriaLigand fishing
Publication Date：2025-04-18
Online Publishing Date：2026-09-14(First online date of this platform, not the publication date of the document)
Pages：4( 535-538 )