小凹蛋白-1改善2型糖尿病小鼠内皮祖细胞血管生成的机制研究  

Mechanisms of caveolin-1 regulating functions of angiogenesis of endothelial progenitor cells in mice with type 2 diabetes

作　　者：黄燕凤 刘玉花[1] 翟露 马翠 戴霞[1] HUANG Yan-feng;LIU Yu-hua;ZHAI Lu;MA Cui;DAI Xia(Department of Nursing,The First Affiliated Hospital of Guangxi Medical University,Nanning,Guangxi 530021,China)

机构地区：[1]广西医科大学第一附属医院护理部,广西南宁530021

出　　处：《现代预防医学》2020年第7期1275-1280,1322,共7页Modern Preventive Medicine

基　　金：国家自然科学基金地区科学基金资助项目(编号:81660147);南宁市青秀区科技计划项目(编号:2018027)。

摘　　要：目的探究小凹蛋白-1(caveolin-1,Cav-1)是否通过调节PI3K/AKT表达改善2型糖尿病小鼠内皮祖细胞(endothelial progenitor cells,EPCs)血管生成能力的机制研究。方法分离培养20只8周龄雄性2型糖尿病小鼠骨髓EPCs,通过免疫荧光染色法鉴定EPCs,并将细胞分为过表达组、沉默表达组、过表达空白载体组、沉默表达空白载体组及对照组。构建Cav-1基因过表达和沉默表达慢病毒载体,分别转染至各组EPCs。qPCR及Western-blotting检测转染后Cav-1、PI3K及AKT蛋白表达情况,并比较各组EPCs血管生成能力。结果Western blot结果显示,过表达组Cav-1表达水平较过表达空白载体组升高(P<0.001),而过表达组PI3K表达水平较过表达空白载体组降低(P<0.001)。沉默表达组Cav-1、PI3K、Phospho-PI3K及AKT的表达水平较沉默表达空白载体组均降低(P<0.05)。EPCs功能检测结果显示,过表达组EPCs形成管状结构数和主干长度均低于过表达空白载体组(均P<0.05);沉默表达组EPCs形成管状结构数和主干长度均低于沉默表达空白载体组(均P<0.05)。结论Cav-1过表达和沉默表达均对EPCs血管生成能力产生抑制作用,PI3K/AKT通路可能参与Cav-1对EPCs功能的调节。Objective To explore mechanism of whether caveolin-1(Cav-1)improve the angiogenic capacity of endothelial progenitor cells(EPCs)mediating the expressions of PI3 K/AKT in mice with type 2 diabetes.Methods Bone marrow EPCs of 208-week-old male type 2 diabetic mice were isolated and cultured,identified by immunofluorescence staining,and the cells were randomly divided into 5 groups,including over-expression group,silenced vector group,over-expression blank vector group,silenced blank vector group and control group.Over-expression and silencing lentivirus vector of Cav-1 gene were constructed and transferred to EPCs of each group.Western blotting was used to detect the expression of Cav-1、PI3 K and AKT proteins in each group after transfection,and the angiogenesis of EPCs in each group was compared.Results The results of western blot showed that the level of Cav-1 in the over-expression group was significantly higher than that in the over-expression blank vector group(P<0.001),while the level of PI3 K in the over-expression vector group was significantly lower than that in the over-expression blank vector group(P<0.001).The levels of the silenced vector group were significantly lower than those of the silenced blank vector group,i.e.,virus-1,PI3 K,phospho-PI3 K and AKT(P<0.05).The results of functional test of EPCs showed that the number of tubular structure and trunk length of EPCs in the over-expression group were lower than those in the over-expression blank vector group(all P<0.05).The number of tubular structures and trunk length of EPCs in the silenced expression group were lower than those in the silenced expression blank vector group(all P<0.05).Conclusion Both over-expression and silencing of Cav-1 inhibited EPCs function,and PI3 K/AKT pathway may be involved in the regulation of Cav-1 on the angiogenic capacity of EPCs.

关 键 词：小凹蛋白-1 内皮祖细胞 PI3K/AKT 2型糖尿病 血管生成能力 

分 类 号：R587.2[医药卫生—内分泌]