基础医学与临床2026，Vol.46Issue(1)：46-55,10.DOI:10.16352/j.issn.1001-6325.2026.01.0046
外源性Ang-1与miR-210共感染的内皮祖细胞腔内移植促进糖尿病慢性下肢缺血血管新生
Endovascular transplantation of endothelial progenitor cells co-infected with exogenous Ang-1 and miR-210 promotes angiogenesis in diabetic chronic lower limb ischemia
摘要
Abstract
Objective To investigate the co-modification of endothelial progenitor cells(EPCs)by angiopoietin-1(Ang-1)and microRNA-210(miR-210).The anti-apoptosis,migration and angiogenesis promotion of endothelial progenitor cells(EPCs)provide experimental basis for the treatment of diabetic chronic lower limb ischemia.Methods EPCs in bone marrow of SD rats were isolated and phenotype was identified by Dil-ac-LDL/FITC-UEA-1 double fluorescence staining microscopy and by flow cytometry(for checking CD34+/CD133+).Ang-1 and miR-210 over-expressed lentiviral vectors were constructed and infected into EPCs,gene expression was de-tected by RT-qPCR.Annexin V/PI staining,Transwell assay and ECMatrix gel angiogenesis assay were used to evaluate cell apoptosis rate,migration and tube formation in medium with high glucose and low oxygen(22.0 mmol/L glucose,3%O2)conditions.The rat hind limb ischemia model was established;And 28 days af-ter intracavity transplantation modified EPCs;Micro-vascular density was detected by CD31 immuno-histochemis-try;And Ang-1/Tie2/PI3K/AKT path-related protein expression was analyzed by Western blot.Results EPCs was successfully cultured and identified.The expression of Ang-1 and miR-210 in the co-transfection group was 3.88 times and 4.21 times higher than those in the control group respectively(P＜0.01).The EPCs apoptosis rate(10.84%)in the co-transfection group was significantly lower than that in the control group(26.22%,P＜0.01).The counting of migrating cells(78.3±5.2)was increased by 3.64 times(P＜0.01).The quantity of vascular branches(15.6±1.8)was increased by 3.7 times(P＜0.01).The microvascular density(97.7±12.5)in the cotransfection group was 3.71 times higher than that in the control group(26.3±18.4,P＜0.05)and the expression of Ang-1,VEGF,P-Akt and P-VEGFR in ischemic tissue was significantly up-regulated(P＜0.05).Conclusions The co-modification of Ang-1 and miR-210 can significantly improve the survival of EPCs in diabetic ischemic micro-environment and promote angiogenesis.Its mechanism is closely related to the activation of Ang-1/Tie2/PI3K/AKT pathway.关键词
内皮祖细胞/血管生成素/微小RNA/慢性下肢缺血Key words
endothelial progenitor cell/angiopoietin/microRNA/chronic lower limb ischemia分类
生物科学引用本文复制引用
李春孟,孙慧艳,郑祥坚,谢尚尚,林德永,刘子田..外源性Ang-1与miR-210共感染的内皮祖细胞腔内移植促进糖尿病慢性下肢缺血血管新生[J].基础医学与临床,2026,46(1):46-55,10.基金项目
浙江省卫生健康委医药卫生科技计划(2020KY295) （2020KY295）
