收稿日期: 2025-11-12
修回日期: 2025-12-01
录用日期: 2025-12-02
网络出版日期: 2026-04-13
基金资助
国家自然科学基金项目(82100067);陕西省自然科学基金项目(2024JC-YBQN-0833)
AMPK activation inhibits pulmonary arterial smooth muscle cells proliferation via YAP/PKM2/glycolysis signaling pathway
Received date: 2025-11-12
Revised date: 2025-12-01
Accepted date: 2025-12-02
Online published: 2026-04-13
目的 明确在肺动脉平滑肌细胞（PASMC）增殖中，Yes相关蛋白（YAP）调控细胞糖酵解的分子机制，并寻找有效干预靶点，旨在为肺动脉高压的临床防治提供潜在靶点及实验依据。 方法 鞘氨醇-1-磷酸（S1P）刺激原代PASMC增殖，Western blot法检测YAP活性以及丙酮酸激酶（PKM2）蛋白水平，Seahorse能量代谢仪检测细胞糖酵解速率，乳酸试剂盒检测培养基中乳酸产量，BrdU掺入法检测细胞增殖情况。 结果 与对照组相比，S1P组的p-YAP水平明显降低，PKM2的蛋白水平明显升高，糖酵解能力和糖酵解容量明显增加，培养基中乳酸的产生量明显升高（P < 0.05）。采用YAP特异性siRNA沉默YAP可逆转S1P诱导的PKM2上调，以及糖酵解能力的增强和乳酸的生成（P < 0.05）。进一步，采用二甲双胍激活AMPK后可抑制YAP活性，进而逆转S1P诱导的PKM2的上调以及糖酵解的增强、乳酸的产生（P < 0.05）。最后，给予YAP siRNA转染、糖酵解抑制剂2-DG以及AMPK激活剂二甲双胍干预可逆转S1P诱导的PASMC增殖（P < 0.05）。 结论 S1P可通过YAP/PKM2/糖酵解信号通路促进PASMC增殖，而二甲双胍激活AMPK后可通过抑制该信号通路，从而抑制PASMC增殖。
柯蕊 , 张伟 , 和平 , 史文花 , 张永红 . 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[J]. 实用医学杂志, 2026 , 42(7) : 1208 -1214 . DOI: 10.3969/j.issn.1006-5725.2026.07.013
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.
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