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目的:探讨二苯乙烯苷(TSG)对糖尿病肾病大鼠肾小球系膜细胞增殖及分泌炎症因子的影响。方法:分别以正常糖(NG)、高糖(HG)、糖基化终产物(AGE)及过氧化氢(H2O2)孵育大鼠肾小球系膜细胞,CCK细胞计数法检测大鼠系膜细胞的OD值,判断细胞增殖情况。比色法检测细胞培养液中的超氧化物歧化酶(SOD)和谷胱甘肽(GSH-Px)的活性、丙二醛(MDA)含量的变化。采用酶联免疫吸附实验(ELISA)检测方法检测培养细胞上清中单核细胞趋化蛋白-1(MCP-1)和细胞间粘附分子-1(ICAM-1)蛋白浓度,Western Blot检测系膜细胞中环氧化酶-2(COX-2)蛋白表达。结果:HG、AGE和H2O2能明显诱导系膜细胞增殖和氧化应激增强,而TSG可明显改善系膜细胞增殖和氧化应激状态,同时三种诱因均可使肾小球系膜细胞MCP-1、ICAM-1和COX-2蛋白分泌增加;TSG预处理后可抑制上述因素诱导的炎症因子分泌。结论:TSG可抑制糖尿病大鼠系膜细胞增殖和调控炎症因子表达,从而在防治糖尿病肾病发生发展过程中发挥重要作用。
Objective: To investigate the effect of stilbene glycoside (TSG) on proliferation and secretion of inflammatory cytokines in glomerular mesangial cells of diabetic nephropathy rats. Methods: Rat mesangial cells were incubated with normal glucose (NG), high glucose (HG), advanced glycation end products (AGE) and hydrogen peroxide (H2O2) Cell OD value, to determine the cell proliferation. The activity of superoxide dismutase (SOD) and glutathione (GSH-Px) and the content of malondialdehyde (MDA) in cell culture medium were detected by colorimetric method. The concentrations of monocyte chemoattractant protein-1 (MCP-1) and intercellular adhesion molecule-1 (ICAM-1) in culture supernatants were detected by enzyme linked immunosorbent assay (ELISA) Membrane cell cyclooxygenase-2 (COX-2) protein expression. Results: HG, AGE and H2O2 could obviously induce mesangial cell proliferation and oxidative stress, while TSG could obviously improve the mesangial cell proliferation and oxidative stress. At the same time, three kinds of inducing factors could make mesangial cells MCP- 1, ICAM-1 and COX-2 protein secretion increased; TSG pretreatment can inhibit the secretion of inflammatory cytokines induced by the above factors. Conclusion: TSG can inhibit the proliferation of diabetic rat mesangial cells and regulate the expression of inflammatory cytokines, which play an important role in the prevention and treatment of diabetic nephropathy.