绞股蓝皂苷对AGEs诱导下人肾小球系膜细胞中RAGE及转化生长因子TGF-β1表达的影响

发布时间：2018-06-04 12:00

本文选题：糖尿病肾病 + 人肾小球系膜细胞　；参考：《桂林医学院》2017年硕士论文

【摘要】：目的:观察绞股蓝皂苷(gypenosides,GP)对晚期糖基化终末产物(advanced glycation endproducts,AGEs)诱导下人肾小球系膜细胞(human mesangial cells,HMCs)中晚期糖基化终末产物受体(RAGE)及转化生长因子-β1(TGF-β1)表达的影响。方法:将体外培养的(含有15%胎牛血清的DMEM低糖培养液)的人肾小球系膜细胞(HMCs)分成四个实验组:正常组、模型组、GP组、阳性对照组。除正常组外,根据前期课题MTT实验结果余下三组均再给予200mg·L-1 AGEs刺激。此外,GP组中再分别加入由低至高三个不同浓度的GP(25、75、175 mg·L-1)进行干预,阳性对照组中加入氨基胍盐酸盐(0.1mmol·L-1)进行干预。同步孵育72小时后,应用Western blotting技术检测各实验组中RAGE和TGF-β1蛋白的表达水平;应用RT-PCR技术检测各实验组中RAGE和TGF-β1 mRNA的表达水平。结果:与正常组比较,体外培养的HMCs,模型组中AGEs诱导下HMCs中RAGE、TGF-β1蛋白及mRNA异常高表达,差异显著(P均0.01);与模型组比较,GP组中GP可明显下调HMCs中RAGE、TGF-β1蛋白及mRNA的表达,并呈浓度依赖性,差异有统计学意义(P均0.01)。结论:GP可降低AGEs诱导下HMCs中RAGE的异常高表达,阻断AGEs-RAGE经典的信号通路,并下调下游转化生长因子-β1的表达活性,进而延缓DN纤维化这一病理进程,对早期DN防治可发挥一定积极地意义。
[Abstract]:Aim: to observe the effect of GypenosidesGypenosidesGPon on the expression of advanced glycation endproducts (ages) and transforming growth factor- 尾 1TGF- 尾 1 (TGF- 尾 1) in human Mesangial cells (HMCs) induced by advanced glycosylation end product receptor (RAGEG) and transforming growth factor 尾 1TGF- 尾 1 (TGF- 尾 1) in human Mesangial cells. Methods: human glomerular Mesangial cells cultured in vitro (DMEM low glucose medium containing 15% fetal bovine serum) were divided into four groups: normal group, model group and positive control group. In addition to the normal group, the other three groups were stimulated with 200mg L -1 AGEs according to the results of the previous MTT experiment. In addition, three different concentrations of GPX 2575175 mg / L were added to GP group and 0.1 mmol / L aminoguanidine hydrochloride was added to positive control group. After 72 hours of incubation, the expression of RAGE and TGF- 尾 1 protein in each experimental group was detected by Western blotting technique, and the expression level of RAGE and TGF- 尾 1 mRNA in each experimental group was detected by RT-PCR technique. Results: compared with the normal group, the expression of RAGETGF- 尾 1 protein and mRNA in HMCs induced by AGEs in the model group was significantly higher than that in the normal group (P 0.01), and the expression of RAGEN TGF- 尾 1 protein and mRNA in HMCs was significantly down-regulated in the GP group compared with the model group. The difference was statistically significant (P < 0.01). Conclusion the AGEs induced abnormal high expression of RAGE in HMCs can be inhibited by AGEs, the classical signal pathway of AGEs-RAGE is blocked, and the activity of transforming growth factor-尾 1 is down-regulated, which can delay the pathological process of DN fibrosis. The prevention and treatment of early DN can play a positive role.
【学位授予单位】：桂林医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R587.2;R692.9

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