S1P/S1PR1通过抑制Pyk2减轻H2O2诱导的血管内皮细胞线粒体损伤

S1P/S1PR1 attenuates H2O2-induced mitochondrial damage in vascular endothelial cells by inhibiting Pyk2

摘要:

目的]探讨1-磷酸鞘氨醇(S1P)是否通过1-磷酸鞘氨醇受体(S1PR)/富含脯氨酸的酪氨酸激酶2(Pyk2)通路调控线粒体钙单向转运体(MCU)表达,从而减少氧化应激诱导的线粒体损伤并抑制线粒体相关凋亡。 [方法]采用过氧化氢(H₂O₂)诱导人脐静脉内皮细胞(HUVEC)氧化损伤模型,并使用不同浓度的S1P处理氧化损伤的HUVEC。同时,实验中应用S1PR1激动剂SEW2871、S1PR1抑制剂W146及Pyk2抑制剂PF-562271,以探讨S1P作用的具体机制。 [结果]S1P处理后,HUVEC受到的氧化损伤明显缓解(P＜0.01),并伴随S1PR1表达的增加(P＜0.05),而S1PR3的表达无显著变化。同时,Pyk2和MCU的表达水平下降(P＜0.05)。SEW2871进一步降低线粒体损伤(P＜0.05),而W146则使损伤加重(P＜0.05)。此外,Pyk2抑制剂PF-562271的应用同样降低了H₂O₂诱导的线粒体损伤(P＜0.05),进一步验证了Pyk2在该过程中发挥的作用。 [结论]S1P通过S1PR1抑制Pyk2的表达,从而减少H₂O₂诱导的HUVEC线粒体损伤并抑制线粒体相关凋亡。

Abstract:

Aim To investigates whether sphingosine-1-phosphate (S1P) regulates the expression of mitochondrial calcium uniporter (MCU) via the sphingosine-1-phosphate receptor/proline-rich tyrosine kinase 2 (S1PR/Pyk2) signaling pathway, thereby reducing oxidative stress-induced mitochondrial damage and inhibiting mitochondria-related apoptosis. Methods Human umbilical vein endothelial cells (HUVEC) were subjected to oxidative damage using hydrogen peroxide (H₂O₂) as a model. Different concentrations of S1P were applied to the oxidative damaged HUVEC. Additionally, the S1PR1 agonist SEW2871, the S1PR1 inhibitor W146, and the Pyk2 inhibitor PF-562271 were used to explore the specific mechanism of S1P action. Results S1P treatment significantly alleviated oxidative damage in HUVEC and was accompanied by an increase in S1PR1 expression (P＜0.05), while S1PR3 expression remained unchanged. Meanwhile, the expression levels of Pyk2 and MCU decreased (P＜0.05). SEW2871 further reduced mitochondrial damage, whereas W146 exacerbated it (P＜0.05). Furthermore, the application of the Pyk2 inhibitor PF-562271 also reduced H₂O₂-induced mitochondrial damage (P＜0.05), further confirming the role of Pyk2 in this process. Conclusion S1P reduces H₂O₂-induced mitochondrial damage and inhibits mitochondria-related apoptosis in HUVEC by suppressing Pyk2 expression via S1PR1.