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目的研究内脂素在人脐静脉内皮细胞(HUVEC)增殖凋亡中的作用及机制。方法选取产科提供的健康剖腹产的孕妇脐带进行原代培养HUVEC。使用MTT法检测内皮细胞增殖,流式细胞仪法检测细胞凋亡情况,通过Western blot检测内皮细胞中磷酸化热休克蛋白27(p-HSP27)的蛋白表达量。内脂素(10nmol/L)分别加入10μmol/L的丝裂原活化蛋白激酶(p38 MAPK),磷脂酰肌醇3激酶(PI3K)或细胞外信号调节激酶(ERK)1/2选择性抑制剂(分别为SB202190,LY294002和U0126)预处理的内皮细胞后,检测内皮细胞增殖、凋亡及蛋白表达情况改变。结果 10nmol/L的内脂素能促进HUVEC的增殖、抑制高糖诱导的凋亡(均P<0.05)。预先给予LY294002或U0126能够有效地减少内脂素诱导的HSP27磷酸化的表达,分别使磷酸化峰值减少77.8%和51.7%(均P<0.01),并且LY294002或U0126可以阻断内脂素诱导的HUVEC的增殖促进及凋亡保护作用。结论内脂素通过ERK1/2和PI3K/蛋白激酶B(Akt)信号通路激活HSP27磷酸化,从而促进HUVEC增殖并抑制高糖诱导凋亡的重要过程。
Objective To study the role and mechanism of visfatin in the proliferation and apoptosis of human umbilical vein endothelial cells (HUVECs). Methods The umbilical cord of pregnant women with obstetrics and gynecology was selected for primary culture of HUVEC. The proliferation of endothelial cells was detected by MTT assay. The apoptosis of cells was detected by flow cytometry. The protein expression of phosphorylated heat shock protein 27 (p-HSP27) in endothelial cells was detected by Western blot. (10 nmol / L) were added 10μmol / L mitogen-activated protein kinase (p38 MAPK), phosphatidylinositol 3 kinase (PI3K) or extracellular signal-regulated kinase (ERK) 1/2 selective inhibitor (Respectively SB202190, LY294002 and U0126) pretreated endothelial cells were detected endothelial cell proliferation, apoptosis and protein expression changes. Results 10 nmol / L of visfatin promoted HUVEC proliferation and inhibited high glucose-induced apoptosis (all P <0.05). Pretreatment with LY294002 or U0126 effectively reduced visfatin-induced phosphorylation of HSP27, which decreased phosphorylation peak by 77.8% and 51.7%, respectively (all P <0.01), and LY294002 or U0126 could block visfatin-induced Proliferation promotion and apoptosis protective effect of HUVEC. Conclusion Visfatin activates phosphorylation of HSP27 through ERK1 / 2 and PI3K / Akt signaling pathways to promote the proliferation of HUVEC and inhibit the apoptosis induced by high glucose.