An Xingxing, Li Lan, Chen Younan, Luo Ai, Ni Zuyao, Liu Jingping, Yuan Yujia, Shi Meimei, Chen Bo, Long Dan, Cheng Jingqiu, Lu Yanrong
Key Laboratory of Transplant Engineering and Immunology, Ministry of Health; West China Hospital, Sichuan University, Chengdu 610041, China.
School of Biomedical Sciences, CHIRI Biosciences, Curtin University, GPO Box U1987, Perth, WA 6845, Australia.
Int J Mol Sci. 2016 Apr 1;17(4):483. doi: 10.3390/ijms17040483.
Glucolipotoxicity is one of the critical causal factors of diabetic complications. Whether mesenchymal stem cells (MSCs) have effects on glucolipotoxicity in human umbilical vein endothelial cells (HUVECs) and mechanisms involved are unclear. Thirty mM glucose plus 100 μM palmitic acid was used to induce glucolipotoxicity in HUVECs. MSCs and HUVECs were co-cultured at the ratio of 1:5 via Transwell system. The mRNA expressions of inflammatory factors were detected by RT-qPCR. The productions of reactive oxygen species (ROS), cell cycle and apoptosis were analyzed by flow cytometry. The tumor necrosis factor-α stimulated protein 6 (TSG-6) was knockdown in MSCs by RNA interference. High glucose and palmitic acid remarkably impaired cell viability and tube formation capacity, as well as increased the mRNA expression of inflammatory factors, ROS levels, and cell apoptosis in HUVECs. MSC co-cultivation ameliorated these detrimental effects in HUVECs, but no effect on ROS production. Moreover, TSG-6 was dramatically up-regulated by high glucose and fatty acid stimulation in both MSCs and HUVECs. TSG-6 knockdown partially abolished the protection mediated by MSCs. MSCs had protective effects on high glucose and palmitic acid induced glucolipotoxicity in HUVECs, and TSG-6 secreted by MSCs was likely to play an important role in this process.
糖脂毒性是糖尿病并发症的关键致病因素之一。间充质干细胞（MSCs）对人脐静脉内皮细胞（HUVECs）的糖脂毒性是否有影响及其相关机制尚不清楚。采用30 mM葡萄糖加100 μM棕榈酸诱导HUVECs发生糖脂毒性。通过Transwell系统将MSCs与HUVECs按1:5的比例共培养。采用RT-qPCR检测炎症因子的mRNA表达。通过流式细胞术分析活性氧（ROS）的产生、细胞周期和细胞凋亡情况。通过RNA干扰敲低MSCs中的肿瘤坏死因子-α刺激蛋白6（TSG-6）。高糖和棕榈酸显著损害HUVECs的细胞活力和管腔形成能力，同时增加其炎症因子mRNA表达、ROS水平及细胞凋亡。与MSCs共培养可改善HUVECs的这些有害影响，但对ROS产生无影响。此外，高糖和脂肪酸刺激均使MSCs和HUVECs中的TSG-6显著上调。敲低TSG-6可部分消除MSCs介导的保护作用。MSCs对高糖和棕榈酸诱导的HUVECs糖脂毒性具有保护作用，MSCs分泌的TSG-6可能在此过程中发挥重要作用。
