论文部分内容阅读
目的观察绞股蓝皂苷(GP)对晚期糖基化终末产物(AGEs)诱导下人肾小球系膜细胞(HMCs)中晚期糖基化终末产物受体(RAGE)表达及氧化应激水平的影响。方法用含13%FBS的DMEM低糖培养液体外培养HMCs细胞,将细胞分为6组:对照组(DMEM培养液),AGEs组(200 mg/L),GP低、中、高剂量(25、75、150 mg/L)组和阳性对照组(氨基胍0.1 mmol/L)。培养72 h后,采用实时荧光定量PCR法检测各组细胞中RAGE m RNA的表达水平,Western blotting法检测RAGE蛋白表达水平。应用试剂盒检测各组细胞上清液中超氧化物歧化酶(SOD)、丙二醛(MDA)及细胞内微量还原型谷胱甘肽(GSH)活性。结果 AGEs显著促进HMCs中RAGE m RNA及蛋白表达(P<0.01),增强细胞氧化应激水平。GP能有效抑制RAGE m RNA及蛋白表达,增加上清液中SOD和细胞内GSH水平,降低细胞上清液中MDA水平,且呈浓度依赖效应,与AGEs组比较差异显著(P<0.01)。结论 GP下调AGEs刺激后HMCs细胞RAGE的异常高表达,同时改善细胞内氧化应激水平,其可能的机制是GP阻断了RAGE介导的AGEs-RAGE-氧化应激信号通路,表现为细胞上清液中MDA水平降低和SOD水平增加及细胞内GSH水平增加。
Objective To investigate the expression of advanced glycation end products receptor (RAGE) and oxidative stress in human glomerular mesangial cells (HMCs) induced by advanced glycation end products (AGEs) of gypenosides influences. Methods HMCs were cultured in vitro with DMEM low glucose medium containing 13% FBS. The cells were divided into 6 groups: control group (DMEM), AGEs group (200 mg / L), low, medium and high dose of GP 75,150 mg / L) group and the positive control group (aminoguanidine 0.1 mmol / L). After cultured for 72 h, the expression of RAGE m RNA in each group of cells was detected by real-time fluorescence quantitative PCR, and the expression of RAGE protein was detected by Western blotting. The activity of superoxide dismutase (SOD), malondialdehyde (MDA) and intracellular micro-reduced glutathione (GSH) in the supernatant of each group were detected by kit. Results AGEs significantly promoted RAGE m RNA and protein expression in HMCs (P <0.01), and increased oxidative stress in cells. GP can effectively inhibit RAGE m RNA and protein expression, increase SOD and intracellular GSH level in supernatant, and decrease MDA in cell supernatant in a concentration-dependent manner, which is significantly different from AGEs group (P <0.01). CONCLUSION GP can down-regulate AGEs-induced abnormal expression of RAGE in HMCs and improve the intracellular oxidative stress. The possible mechanism is that GP blocks RAGE-mediated AGEs-RAGE-oxidative stress signaling pathway, Serum MDA levels decreased and SOD levels increased and intracellular GSH levels increased.