2型糖尿病来源外泌体转运circ_001895对高糖胁迫下内皮祖细胞功能的影响

Effect of Exosomal Transport of circ_001895 from Type 2 Diabetes Mellitus on Endothelial Progenitor Cells under High Glucose Stress

目的:探讨2型糖尿病(T2DM)来源的外泌体(EXO)中转运环状RNA_001895(circ_001895)对高糖胁迫下内皮祖细胞的增殖、凋亡、迁移及血管生成标志物的影响。方法:从T2DM患者中获取血清样本,分为血清组和EXO组。采用RT-qPCR技术检测血清和EXO中circ_001895的表达水平。将T2DM患者血清来源的EXO中circ_001895表达显著升高的EXO命名为EXO-circ。使用武汉普诺赛公司提供的内皮祖细胞进行培养,并分为以下3组:Ctrl组:常规培养24h,不进行额外处理。高糖(HG)组:以30mmoL/L葡萄糖处理细胞24h。HG+EXO-circ组:以30mmoL/L葡萄糖联合100μg/mL的EXO-circ处理细胞24h。采用RT-qPCR技术检测各组细胞中circ_001895的表达。使用CCK-8法检测细胞增殖活力。通过流式细胞术检测细胞凋亡情况。利用Western blot技术检测血管生成标志物E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)和波形蛋白(Vimentin)的表达。通过Transwell细胞迁移实验检测细胞的迁移能力。结果:与血清组相比,EXO组中circ_001895的相对表达量显著上调(P<0.05)。与Ctrl组相比,HG组中circ_001895和E-cadherin的相对表达量及细胞凋亡率均显著上调(均P<0.05),而细胞增殖活力、迁移能力以及N-cadherin和Vimentin的相对表达量均显著下调(均P<0.05)。与HG组相比,HG+EXO-circ组中circ_001895和E-cadherin的相对表达量及细胞凋亡率均显著上调(均P<0.05),而细胞增殖活力、迁移能力以及N-cadherin和Vimentin的相对表达量均显著下调(均P<0.05)。结论:T2DM来源的EXO转运circ_001895抑制高糖胁迫下内皮祖细胞的增殖、迁移以及血管生成,并促进细胞的凋亡。

Objective:To investigate the effects of exosomal circ_001895 transportation on endothelial progenitor cells under high glucose stress derived from patients with type 2 diabetes(T2DM).Methods:Serum samples were obtained from T2DM patients and divided into serum group and EXO group.The expression levels of circ_001895 in serum and EXO were detected by using RT-qPCR technology.EXOs derived from T2DM patients'serum with significantly elevated circ_001895 expression were named EXO-circ.Endothelial progenitor cells provided by Wuhan Procell were cultured and divided into the following three groups.Control Group:Cultured conventionally for 24 hours without additional treatment.High Glucose(HG)Group:Treated with 30 mmoL/L glucose for 24 hours.HG+EXO-circ Group:Treated with 30 mmoL/L glucose combined with 100μg/mL EXO-circ for 24 hours.The expression of circ_001895 in each group of cells was detected using RT-qPCR technology.Cell proliferation vitality was detected using the CCK-8 method.Cell apoptosis was detected using flow cytometry.The expression of angiogenic markers E-cadherin,N-cadherin,and Vimentin was detected using Western blot technology.Cell migration capacity was detected using the Transwell cell migration assay.Results:Compared to the serum group,the relative expression level of circ_001895 in the EXO group was significantly upregulated(P<0.05).Compared to the control group,the relative expression levels of circ_001895 and E-cadherin and the cell apoptosis rate in the HG group were significantly upregulated(all P<0.05),while cell proliferation vitality,migration capacity,and the relative expression levels of N-cadherin and Vimentin were significantly downregulated(all P<0.05).Compared to the HG group,the relative expression levels of circ_001895 and E-cadherin and the cell apoptosis rate in the HG+EXO-circ group were significantly upregulated(all P<0.05),while cell proliferation vitality,migration capacity,and the relative expression levels of N-cadherin and Vimentin were significantly downregulated(all P<0.05).Conclusion:Transportation of circ_001895 in exosomes from T2DM inhibits proliferation,migration,and vascular generation of endothelial progenitor cells under high glucose stress,while promoting apoptosis.