红景天苷对间充质干细胞缺氧损伤后增殖及抗氧化能力影响的初步研究

摘要

Objective] To investigate the influence of Salidroside impact on proliferation and anti-oxidant ability of mesenchymal stem cells(MSCs) against hypoxia. [Method] Rat bone marrow MSCs were cultured in vitro and the adipogenic differentiation and osteogenic differentiation potential were identified. Hypoxia cell model was established by 100μmol·L-1 cobalt chloride, and then cells were divided into normal control group, hypoxia group, and Salidroside groups(30, 60, 120, 240, 480μmol·L-1 Salidroside). Methyl thiazolyl tetrazolium method was used to detect the proliferation of cells, and the lactate dehydrogenase(LDH) level in cultural supernatants and cell superoxide dismutase(SOD) activity were detected according to the kit methods. [Result] After adipogenic differentiation, MSCs appeared orange-red spherical lipid droplets by oil red O staining, and silver staining was positive after osteogenic differentiation, what confirmed MSCs could differentiate to adipocyte and osteoblast. The proliferation of MSCs in hypoxia group was lower than normal control group(P<0.01), and the proliferation of MSCs in Salidroside groups was higher than hypoxia group( P<0.05,P<0.01). The LDH level of cell supernatants in Salidroside groups was lower than hypoxia group(P<0.05,P<0.01), and the SOD activity of MSCs in Salidroside groups was higher than hypoxia group(P<0.05,P<0.01), with concentration increasing. [Conclusion] Salidroside plays a significant role in the protection on hypoxic injured mesenchymal stem cells induced by cobalt chloride, and promotes the proliferation, the mechanism may be related to the enhancement of cellular anti-oxidative capacity.%[目的]研究红景天苷对体外培养的间充质干细胞（mesenchymal stem cells，MSCs）缺氧损伤后增殖及抗氧化能力的影响。[方法]体外培养大鼠骨髓MSCs，检测其成脂、成骨分化能力。采用100μmol·L-1氯化钴建立体外培养的MSCs缺氧损伤模型。将细胞分为正常对照组、缺氧损伤组、不同浓度红景天苷组（30、60、120、240、480μmol·L-1）。四甲基偶氮唑蓝（methyl thiazolyl tetrazolium，MTT）法检测各组细胞增殖能力。同时，比较各组细胞上清液中乳酸脱氢酶（lactate dehydrogenase，LDH）水平、细胞中超氧化物歧化酶（superoxide dismutase，SOD）活性。[结果] MSCs成脂诱导后，经油红O染色出现橙红色的圆球形脂滴；MSCs成骨诱导后，硝酸银染色呈阳性，证实培养的MSCs具有成脂、成骨分化能力。缺氧损伤组MSCs的增殖能力较正常组降低（P<0.01），红景天苷组随浓度的增加对MSCs的增殖能力较缺氧损伤组均有不同程度的提高（P<0.05，P<0.01）。红景天苷组随浓度的增加细胞上清液中LDH量较缺氧损伤组均有不同程度的减少（P<0.05，P<0.01），红景天苷组随浓度的增加细胞内SOD活性较缺氧损伤组均有不同程度的提高（P<0.05，P<0.01）。[结论]红景天苷对氯化钴诱导的MSCs缺氧损伤具有保护作用，可促进细胞增殖，其机制与增强细胞抗氧化能力有关。