AMPK调控YAP/PKM2/糖酵解信号通路抑制肺动脉平滑肌细胞增殖的机制OA

Objective To investigate the molecular mechanism by which Yes-associated protein(YAP)regulates glycolysis during pulmonary arterial smooth muscle cell(PASMC)proliferation,and to identify potential therapeutic targets.Methods Primary cultured PASMC were stimulated with sphingosine-1-phosphate(S1P).YAP phosphorylation and pyruvate kinase(PKM2)protein expression were determined by Western blot.Cellular glycolytic function was assessed by measuring the glycolysis rate with a seahorse metabolic analyzer and quantifying lactate production using a lactate assay kit.Cell proliferation was analyzed by a BrdU incorporation assay.Results Compared with the control group,the S1P group showed significantly reduced p-YAP level,increased PKM2 protein expression,enhanced glycolytic capacity,and elevated lactate production in the culture medium(P＜0.05).Knockdown of YAP with specific siRNA attenuated S1P-induced PKM2 upregulation,as well as the enhancement of glycolytic capacity and lactate production(P＜0.05).Furthermore,activation of AMPK by metformin inhibited YAP activity,thereby reversing S1P-induced upregulation of PKM2,enhanced glycolysis,and lactate production(P＜0.05).Finally,interventions with YAP siRNA transfection,the glycolysis inhibitor 2-DG,and the AMPK activator metformin mitigated S1P-induced PASMC proliferation(P＜0.05).Conclusion S1P promotes PASMC proliferation through the YAP/PKM2/glycolysis signaling pathway,while metformin activates AMPK to inhibit this pathway,thereby suppressing PASMC proliferation.