间充质干细胞外泌体对腹膜间皮细胞高糖损伤的作用研究

Effect of mesenchymal stem cells-derived exosomes on hyperglycemic injury of human peritoneal mesothelial cells

目的 探讨间充质干细胞来源外泌体(MSCs-Exo)对高糖致腹膜间皮损伤的影响。方法 将MET-5A细胞分成3组:正常对照组(不予处理)、高糖组(高糖处理)、共培养组(高糖和MSCs-Exo共培养),通过CCK-8法选用效果最明显的高糖浓度和MSCs-Exo浓度,各组分别处理24 h后收集细胞和上清液。CCK-8法测定各组细胞增殖活力,流式细胞技术检测细胞凋亡情况,qRT-PCR检测细胞因子IL-6、TNF-α的表达,ELISA检测培养上清液中IL-6的水平。结果高糖组细胞的增殖能力明显低于正常对照组(P<0.001),细胞凋亡率、IL-6和TNF-α的表达显著高于正常对照组(P<0.001);与高糖组相比,共培养组细胞增殖能力提高(P<0.001),细胞凋亡率、IL-6和TNF-α的表达则明显下调(P<0.05)。结论 高糖对腹膜间皮细胞有明显的促凋亡和促炎作用,MSCs-Exo可以促进腹膜间皮细胞的增殖,抑制高糖刺激下细胞凋亡的发生和炎症因子的表达。

Objective To explore the effect of mesenchymal stem cells derived exosomes(MSCs-Exo) on peritoneal mesothelial injury induced by high glucose. Methods MET-5A cells were divided into three groups: normal control group(no treatment), high glucose group(treated with high glucose) and co-culture group(treated with high glucose and MSCsExo). High glucose concentration and MSCs-Exo concentration with the best stimulation effect were selected by CCK-8method. After 24 hours of treatment in each group, the cells and supernatant were collected. The cells proliferation activity was determined by CCK-8, cell apoptosis was tested by flow cytometry, the expression of cytokines IL-6 and TNF-α was detected by qRT-PCR, and the level of IL-6 in culture supernatant was detected by ELISA. Results The cell viability of high glucose group was significantly lower than that of normal control group(P<0.001), and the apoptosis rate, expression of IL-6 and TNF-α were significantly higher(P<0.001). Compared with the high glucose group, the cell viability of the coculture group was increased(P<0.001), and the apoptosis rate, IL-6 and TNF-α expression were significantly down-regulated(P <0.05). Conclusion High glucose has significant pro-apoptotic and pro-inflammatory effects on mesothelial cells. While MSCs-Exo can promote mesothelial cells proliferation and inhibit the occurrence of apoptosis and the expression of inflammatory factors caused by high glucose.