尿源性干细胞外泌体对高糖诱导内皮细胞损伤的保护作用

Exosomes secreted by urine-derived stem cells inhibits high glucose-induced endothelial dysfunction

目的探究尿源性干细胞外泌体(urine-derived stem cell exosomes,USC-Exo)对高浓度葡萄糖诱导的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)损伤模型的保护作用。方法采用酶灌注法、超速离心法获取HUVEC和USC-Exo。以高糖诱导的HUVEC损伤模型为研究对象,设立空白对照组、造模组和给药实验组。采用CCK-8测定各组细胞增殖活力,挑选最适合给药浓度;观察药物影响下各组细胞在划痕迁移、成管实验以及低密度脂蛋白(low-density lipoprotein,LDL)摄取中的表现差异;采用qRT-PCR方法检测B细胞淋巴瘤因子-2(B-cell lymphoma-2,Bcl-2)、Bcl-2相关X蛋白(Bcl-2 associated X protein,BAX)、超氧化物歧化酶2(superoxide dismutase-2,SOD2)等基因的表达。结果本实验获取了高质量的HUVEC以及USC-Exo;在高糖影响下内皮细胞的增殖活力、迁移成管能力、摄取LDL的功能相比对照组均有所下降(P<0.05),而USC-Exo则可以实现逆转,优化内皮细胞功能,逆转凋亡基因的表达,增强抗氧化能力(P<0.05)。结论USC-Exo可在一定程度上保护内皮细胞,缓解高糖造成的细胞损伤。

Objective To explore the protective effects of human urine-derived stem cell exosomes(USC-Exo)on high glucose-induced injury in human umbilical vein endothelial cells(HUVEC).Methods HUVEC and USC-Exo were obtained by enzyme perfusion and ultracentrifugation respectively.A high glucose-induced HUVEC injury model was adopted in the research.HUVEC was divided into 3 groups:control group,model group and treatment group.CCK-8 was used to determine the cell proliferation activity of each group to select the most suitable drug concentration.We observed differences in the performance of each group cells in scratch migration,tube formation experiment,and low-density lipoprotein(LDL)ingestion under the influence of drugs.qRT-PCR was used to detect the expression of genes such as B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(BAX)and superoxide dismutase-2(SOD2).Results We obtained high-quality HUVEC and USC-Exo in this experiment.Under the influence of high glucose,the proliferation activity,angiogenic capacity and the function of ingesting LDL in model group were decreased compared with the control group(P<0.05).While USC-Exo could reverse these phenomena,optimize endothelial cell function,reduce the expression of apoptotic genes,and enhance antioxidant capacity(P<0.05).Conclusion USC-Exo can protect endothelial cells to a certain extent and alleviate cell damage caused by high glucose.