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

doi:10.3969/j.issn.1006-5725.2026.07.013

摘要/Abstract

摘要：

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。

关键词: 细胞增殖, Yes相关蛋白, AMPK, 糖酵解

Abstract:

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.

Key words: proliferation, Yes-associated protein, AMP-activated protein kinase, glycolysis

中图分类号:

R543.2

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.

Rui KE,Wei ZHANG,Ping HE,Wenhua SHI,Yonghong ZHANG. AMPK activation inhibits pulmonary arterial smooth muscle cells proliferation via YAP/PKM2/glycolysis signaling pathway[J]. The Journal of Practical Medicine, 2026, 42(7): 1208-1214.

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组别	糖酵解能力	糖酵解容量	乳酸产生量
对照组	17.86 ± 0.68	37.68 ± 3.18	1.00 ± 0.16
S1P组	32.06 ± 1.79^*	73.80 ± 4.45^*	1.89 ± 0.25^*
t值	-12.84	-11.446	-7.324
P值	0.00	0.00	0.00

组别	糖酵解能力	糖酵解容量	乳酸产生量
对照组	15.32 ±2.71	41.21 ± 0.91	1.00 ± 0.15
S1P组	35.20 ± 3.56^*	77.11 ± 2.70^*	1.80 ± 0.23^*
con siRNA + S1P组	33.12 ± 5.32^*	74.09 ± 2.51^*	1.76 ± 0.19^*
YAP siRNA + S1P组	20.99 ± 2.49^#	48.37 ± 2.38^#	1.17 ± 0.17^#
F值	20.12	194.58	27.29
P值	0.00	0.00	0.00

组别	糖酵解能力	糖酵解容量	乳酸产生量
对照组	15.68 ±0.08	40.60 ± 2.51	1.00 ± 0.05
S1P组	31.41 ± 5.89^*	81.46 ± 2.72^*	1.81 ± 0.18^*
MET组	14.70 ± 0.97	39.75 ± 1.76	0.94 ± 0.04
S1P + MET组	20.25 ±1.54^#	45.00 ± 0.90^#	1.14 ± 0.22^#
F值	81.99	271.84	45.45
P值	0.00	0.00	0.00