The purpose of the present research was to discover the function of kaempferol (KAE) in high glucose (HG)-induced glomerular mesangial cells (GMCs) and the underlying mechanism.The HG model of GMCs was established in vitro.Cell proliferation,reactive oxygen species (ROS) generation,NADPH oxidase (NOX),superoxide dismutase (SOD),malondialdehyde (MDA) were studied using commercial kits.The expression of related factors in GMCs were detected by qRT-PCR and Western blot.siNOX4,sip22phox and compound C were detected the effects of KAE on GMCs intracellular signaling pathway induced by HG with Western blot.The interaction of KAE with Sestrin2 and AMPK wered studied by molecular docking method.Results demonstrated that KAE significantly alleviated cell proliferation,reduced ROS,NOX,and MDA levels,meanwhile enhance SOD activity in HG-induced GMCs.Besides,the production of TGF-β1,Collagen IV (Col IV),NOX4,and p22phox were also inhibited by KAE.In addition,KAE elevated the expression levels of Sestrin2 and AMPK in HG-induced GMCs.siNOX4 and sip22phox suppressed HG-induced ROS,TGF-β1,and Col IV production,indicating that it might be mediated by NOX4/p22phox signaling.Compound C reversed the protective effects of KAE towards HG-induced cell proliferation,ROS,NOX4,TGF-β1,and Col IV in GMCs.These findings revealed that KAE inhibited HG-induced OS and ECM accumulation in GMCs,which is partially mediated via AMPK/NOX4 pathway.