斯钙素1激活PI3K/Akt/eNOS通路促进内皮祖细胞增殖和迁移  

Stanniocalcin 1 promotes proliferation and migration of endothelial progenitor cells via PI3K/Akt/eNOS pathway activation

作　　者：袁方正圆 杨杰[1] 王彦伟[1] 张弢[1] 黄岚[1] 

机构地区：[1]第三军医大学新桥医院全军心血管病研究所,重庆400037

出　　处：《第三军医大学学报》2016年第13期1475-1480,共6页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81370211)~~

摘　　要：目的研究斯钙素1(stanniocalcin 1,STC1)对内皮祖细胞(endothelial precusor cells,EPCs)增殖和迁移的影响并探讨其机制。方法密度梯度离心法分离、培养后,采用荧光双染法鉴定SD大鼠脾源性EPCs。不同浓度人重组斯钙素1(recombinant human stanniocalcin 1,rhSTC1)处理EPCs,结合siRNA技术干扰EPCs STC1表达,检测EPCs增殖能力及迁移能力的变化;观察STC1对蛋白激酶B(protein kinase B,PKB/Akt)、内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)及其磷酸化水平的影响;然后观察使用磷脂酰肌醇3激酶(phosphatidylinositol 3-kinase,PI3K)特异性抑制剂LY294002和eNOS特异性抑制剂L-NAME预处理的EPCs对rhSTC1的反应。结果荧光双染法鉴定EPCs阳性率约为88.5%;外源性给予rhSTC1呈浓度依赖性的促进EPCs增殖与迁移;而干扰内源性STC1的表达,则抑制其增殖与迁移;外源性给予rhSTC1增加EPCs中Akt和eNOS的磷酸化水平,而运用PI3K抑制剂(LY294002)和eNOS抑制剂(L-NAME)分别处理EPCs后,rhSTC1诱导的EPCs增殖与迁移能力明显降低。结论 STC1通过激活PI3K/Akt/eNOS信号通路,增强EPCs增殖、迁移能力,为损伤血管的修复提供了新的靶点。Objective To determine the effects of stanniocalcin 1 （STC1） on the proliferation and migration of endothelial progenitor cells （EPCs）, and investigate the potential mechanism involved. Methods EPCs were isolated from the spleens of Sprague-Dawley rats by density-gradient centrifugation, and then cultured and identified with double-fluorescent immunostaining. These EPCs were treated with recombinant human STC1 at different concentrations, or transfeeted with its siRNA to silence the STC1 expression. The proliferation and migration properties of these EPCs as well as the effects of STC1 on the expression levels of protein kinase B （Akt）, endothelial nitric oxide synthase （eNOS） and their phosphorylation were detected. Selective phosphatidyl inositol 3 kinase （PI3K） inhibitor （LY294002） and eNOS inhibitor （L-NAME） were used to reverse the response of pre-treated EPCs to rhSTC1. Results Double-fluorescent immunostaining indicated that the purity of our EPCs was about 88. 5%. Exogenous rhSTC1 treatment promoted the proliferation and migration of EPCs in a dose-dependent manner via the PI3K/Akt/eNOS pathway activation, which was reflected by the increasing of Akt and eNOS phosphorylation levels. Endogenous STCl-expression silence interfered the proliferation and migration of EPCs, and inhibitors LY294002 and L-NAME inhibited the rhSTCl-induced effect on pre-treated EPCs by blocking PI3K/Akt/eNOS pathway. Conclusion STC1 enhances EPCs to proliferate and migrate by activating PI3K/Akt/eNOS pathway. Our findings maybe provide a new therapeutic target for the repair of injured vascular endothelium.

关 键 词：斯钙素1 内皮祖细胞 增殖 迁移 蛋白激酶B 内皮型一氧化氮合酶 

分 类 号：R322.12[医药卫生—人体解剖和组织胚胎学] R329.28[医药卫生—基础医学]