AMPK-mediated Anti-pulmonary Fibrosis Effect Of Adiponectin And JXD

AMPK-mediated energy metabolism pathway plays an important role in regulating pulmonary fibrosis.Adiponectin?ADPN?is an important adipocytokine that participates in the regulation of energy metabolism in the body and has been reported to have anti-fibrotic effects.JXD is an AMPK agonist with a completely new structure.This thesis intends to investigate the basis of the anti-pulmonary fibrosis effect of the endogenous substance adiponectin and the exogenous small molecule JXD,and explore whether its mechanism is related to the activation of AMPK-mediated energy metabolism pathways.Four parts were as follows.?1?Mechanism research of Anti-pulmonary fibrosis effect of ADPN:?1?Animal experiment:The lung fibersis model of C57BL/6 mice and Adipo^-/-mice were successfully established by injection of bleomycin via ntravenous route within 14 days.For C57BL/6 mice,the expression level of p-AMPK in lung tissue was significantly reduced and the content of TGF-?1 increased.Treatment with exogenous ADPN significantly improved the above symptoms induced by BLM.Conversely,adipo^-/-mice exhibited more severe alveolar inflammation and fibrosis than C57BL/6 mice,which showed lower levels of AMPK phosphorylation and higher levels of TGF-?1.Compound C,an AMPK inhibitor,can reverse the protective effect of exogenous ADPN on BLM-induced lung fibrosis in C57BL/6 mice.The results indicate that ADPN may play an anti-pulmonary fibrosis mediated by AMPK.?2?Cell experiment:In the model of lung fibroblast HLF-1 cell proliferation induced by TGF-?1,HLF-1 cell proliferation can be significantly inhibited by ADPN with the increase the expression of p-AMPK protein,reduce the expression of connective tissue growth factor?CTGF?,alpha smooth muscle actin??-SMA?and collagen production.Administration of Compound C,an AMPK inhibitor,can reverse the above effects of ADPN on TGF-?1.These results suggested that ADPN may inhibit TGF-?1/CTGF pathway by activating AMPK phosphorylation to reduce cell proliferation and extracellular matrix?ECM?production.Study on anti-pulmonary fibrosis of small molecular compound JXD:?1?Animal experiment was carry out to discuss the effect of small molecule compound JXD on bleomycin-induced pulmonary fibrosis in mice.The lung function of pulmonary fibrosis mice which induced by bleomycin was significantly improve after administration of JXD,which showed obvious dose correlation and PFD.Meanwile,alveolar inflammation and degree of fibrosis has been relieved.The activity of AMPK was increased and the concentration of TGF-?1 decreased.After administration of Compound C,an AMPK inhibitor,the above effects of JXD were reversed.?2?Cell experiment was carry out to research the effect of small molecule compound JXD on TGF-?1 induced human lung fibroblast HLF-1 cell proliferation model in vitro.The proliferation and activation of HLF-1 cells?induced by TGF-?1?and the expression of a-SMA and CTG were significantly inhibited after treating with JXD and PFD.Meanwile,the phosphorylation level of AMPK was significantly promoted.After administration of Compound C?4?M?,an AMPK inhibitor,the above effects of JXD were reversed.In summary,we demonstrate that both adiponectin and small molecule compound JXD protected against pulmonary fibrosis in BLM-treated mice and inhibited fibrogenic response in TGF-?1-induced lung fibroblasts.The mechanism is related to the inhibition of the TGF-?1/CTGF pathway through the activation of AMPK by adiponectin or JXD,which resulting in the reduction of lung fibroblast proliferation and collagen production.This study is expected to provide new ideas for the development of drugs for the treatment of pulmonary fibrosis.