摘要
目的:探讨甘草苷(LIQ)上调丙酮酸激酶M2(PKM2)表达对过氧化氢(H2O2)诱导人脐静脉内皮细胞(HUVECs)血管新生的影响及可能机制.方法:使用H2O2诱导HUVECs损伤,将细胞分为对照组、H2O2组、H2O2+LIQ组、H2O2+LIQ+Shikonin(PKM2抑制剂)组、H2O2+LIQ+Stattic(STAT3抑制剂)组.采用细胞划痕及管腔形成实验检测细胞迁移、成管能力,Western Blot检测血管新生关键蛋白VEGF、FGF2的表达,以及PKM2、磷酸化PKM2(p-PKM2)和PKM2下游激酶STAT3、p-STAT3的蛋白表达.结果:与H2O2组相比,LIQ处理显著提高了细胞迁移、成管能力,并增加VEGF、FGF2和PKM2、p-PKM2的表达水平;LIQ上调PKM2表达,显著增加了STAT3、p-STAT3表达水平.与H2O2+LIQ组相比,抑制PKM2或STAT3后细胞迁移、成管能力显著下降,VEGF、FGF2、STAT3及p-STAT3蛋白升高趋势被逆转.结论:LIQ可通过上调PKM2表达激活STAT3信号通路促进H2O2诱导的HUVECs血管新生.
Abstract
Objective:To investigate whether liquiritin(LIQ)up-regulation of pyruvate kinase M2(PKM2)on the angiogenesis of human umbilical vein endothelial cells(HUVECs)induced by hydrogen peroxide(H2O2)and its possible mechanism.Methods:HUVECs were damaged by H2O2,and the cells were di-vided into the control group,H2O2 group,H2O2+LIQ group,H2O2+LIQ+Shikonin(PKM2 inhibi-tor)group,and H2O2+LIQ+Static(STAT3 inhibitor)group.Cell migration and tubular formation ability were detected by cell scratch and lumen formation assay.Western Blot was used to detect the ex-pression of key angiogenic proteins VEGF and FGF2,and the expression of proteins,PKM2 and phos-phorylationp-PKM2(p-PKM2),and PKM2 downstream kinase STAT3/p-STAT3.Results:Com-pared with the H2O2 group,LIQ treatment significantly improved cell migration and tubeforming abili-ty,and the expression levels of VEGF,FGF2,PKM2,and p-PKM2.LIQ up-regulated PKM2 ex-pression and significantly increased the expression levels of STAT3 and p-STAT3.Compared with those in the H2O2+LIQ group,cell migration,and tube-forming ability were significantly decreased af-ter PKM2 or STAT3 inhibition,and the increasing trend of VEGF,FGF2,STAT3,and p-STAT3 proteins was reversed.Conclusion:LIQ can activate the STAT3 signaling pathway by upregulating PKM2 expression to promote H2O2-induced angiogenesis in HUVECs.
基金项目
国家自然科学基金资助项目(81570333)
国家自然科学基金资助项目(81270271)
国家自然科学基金青年基金资助项目(30900609)
中央高校基本科研业务费专项资金(2042020kf1014)
NETL
NSTL
万方数据