高糖对滑膜间充质干细胞衰老的影响

Impact of high glucose on senescence of rat synovial mesenchymal stem cells

目的探究高糖对大鼠滑膜间充质干细胞(synovial mesenchymal stem cells,SMSCs)衰老的影响,为SMSCs应用于糖尿病患者骨关节炎的治疗提供风险评估依据。方法葡萄糖生理浓度(5.5 mmol/L)作为对照组,高糖(25 mmol/L)作为高糖组,检测大鼠SMSCs在高糖刺激下的衰老情况,包括细胞增殖活性,β-半乳糖苷酶(SA-β-gal)表达,衰老相关表型(SASP)IL6、MMP13、TNFα、CXCL1、IFNβ的mRNA表达,细胞内线粒体裂解情况,p21蛋白表达,以及透明质酸的分泌。结果本实验条件下,高糖刺激不影响大鼠SMSCs增殖(P=0.675),SA-β-gal染色阳性细胞增加,SMSCs胞内线粒体裂解增加,线粒体呈碎片化,衰老相关表型SASP的mRNA表达增加,MMP13(P=0.011),TNF、IL-6、INFβ(P=0.000),CXCL1(P=0.003),p21蛋白表达下降(P=0.00),透明质酸释放降低(P=0.002)。结论高糖使大鼠SMSCs进入衰老状态,损害其生物学功能,采用SMCSs治疗糖尿病患者骨关节炎其临床效果有待商榷。

Objective To investigate the impact of high glucose on senescence of rat synovial mesenchymal stem cells(SMSCs)so as to provide a reference for risk management of SMSCs in the treatment of osteoarthritis in diabetic patients.Methods The rat SMSCs were divided into control group(physiological concentration of glucose with 5.5 mmol/L)and treatment group(high glucose with 25 mmol/L).The senescence of rat SMSCs was detected,including cell proliferation,SA-β-Gal staining,mRNA expression of SASP(IL-6,MMP-13,TNF-α,CXCL1,IFN-β),intracellular mitochondrial damage,p21 protein expression,and hyaluronic acid release.Results The high glucose medium did not affect the proliferation of SMSCs in the rats(P=0.675).Also,the high glucose medium enhanced the number of SA-β-Gal staining positive cells,the fragmentation of intracellular mitochondrial,and the mRNA expression of SASP,namely MMP-13(P=0.011),TNF,TL-6,INFβ(P=0.000),CXCL1(P=0.003).Meanwhile,the Western blot and ELISA results demonstrated significant down-regulated expressions of P21(P=0.00)and hyaluronic acid(P=0.002).Conclusion High glucose induces senescence and biological function impairment of SMSCs in rats.The clinical effect of SMCSs in the treatment of osteoarthritis in diabetic patients remains to be confirmed.