青蒿琥酯对高糖环境下大鼠肾小球系膜细胞增殖、凋亡及分泌炎症因子TNF-α、IL-6的影响

发布时间：2018-05-09 14:56

本文选题：青蒿琥酯 + 肾小球系膜细胞　；参考：《桂林医学院》2017年硕士论文

【摘要】：目的:观察高糖环境下大鼠肾小球系膜细胞增殖的情况,探讨不同浓度青蒿琥酯是否可抑制高糖环境培养的大鼠肾小球系膜HBZY-1细胞增殖和炎症因子TNF-α、IL-6的分泌、并诱导其凋亡。方法:将大鼠肾小球系膜细胞分成7组:正常培养对照组、高糖培养模型组、青蒿琥酯微剂量组、青蒿琥酯小剂量组、青蒿琥酯中剂量组、青蒿琥酯大剂量组、依那普利阳性药物对照组。倒置显微镜下观察各组肾小球系膜细胞形态变化。采用MTT法观察青蒿琥酯对各组肾小球细胞细胞24h、48h、72h增殖的抑制情况,并通过流式细胞仪检测各组细胞凋亡率,ELISA法观察各组细胞上清液中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)的分泌量变化。结果:(1)青蒿琥酯细胞12h左右贴壁,24h伸展成纺锤状或不规则形状,48h进入对数期生长,3-4天可长至融合。细胞核成卵圆形或圆形。加入药物后,各组细胞融合度有差异。(2)MTT结果显示:正常培养对照组细胞增殖抑制率明显高于高糖培养模型组(P0.01),各浓度青蒿琥酯药物组的抑制率均高于高糖组(P0.05),并成一定时间、剂量依赖性。与依那普利组对比,青蒿琥酯微剂量组抑制率无统计学意义(P0.05)。(3)青蒿琥酯对HBZY-1细胞凋亡有诱导作用(P0.05),并呈一定剂量依赖性;(4)ELISA结果显示:与正常对照组相比,高糖培养模型组细胞培养的上清液中TNF-α、IL-6的含量明显增高(P0.01);与高糖培养模型组比较,各青蒿琥酯药物组的上清液中TNF-α、IL-6的含量均降低(P0.05),大剂量青蒿琥酯组比依那普利组作用明显(P0.01)。结论:高糖环境可促进HBZY-1细胞增殖和炎症因子TNF-α、IL-6的分泌,可诱导机体炎症反应;青蒿琥酯能明显逆转高糖刺激下GMCs的增殖和凋亡并且对高糖环境下HBZY-1细胞分泌TNF-α、IL-6有明显抑制作用。由此推论青蒿琥酯可以作为治疗、防治糖尿病肾病的有效药物,并减轻高糖环境对组织器官的损害。
[Abstract]:Aim: to observe the proliferation of rat glomerular Mesangial cells in high glucose environment, and to investigate whether artesunate at different concentrations can inhibit the proliferation of glomerular Mesangial HBZY-1 cells and the secretion of inflammatory factor TNF- 伪 and IL-6, and induce apoptosis in rat glomerular Mesangial HBZY-1 cells cultured in high glucose environment. Methods: rat glomerular Mesangial cells were divided into 7 groups: normal culture control group, high glucose culture model group, artesunate microdose group, artesunate small dose group, middle dose group of artesunate, high dose group of artesunate. Enalapril positive drug control group. The morphological changes of glomerular Mesangial cells were observed under inverted microscope. The inhibitory effects of artesunate on proliferation of glomerular cells at 24 h and 48 h and 72 h were observed by MTT assay. The expression of TNF- 伪 TNF- 伪 and IL-6 in the supernatant of each group was detected by flow cytometry and Elisa. Results (1) artesunate cells extended into spindle-like or irregularly shaped cells for 12 h or so for 24 h. The cells could grow to fusion in 3 to 4 days after entering logarithmic phase. The nucleus becomes oval or round. The results of MTT showed that the inhibition rate of cell proliferation in normal culture group was significantly higher than that in high glucose culture model group (P 0.01), and the inhibition rate of artesunate drug group in each concentration was higher than that in high glucose group, and the inhibition rate of artesunate drug group was higher than that of high glucose group, and the inhibition rate of artesunate group was higher than that of high glucose group. Dose dependence. In comparison with enalapril group, the inhibitory rate of artesunate microdose group was not statistically significant (P 0.05. 0. 3) artesunate induced apoptosis of HBZY-1 cells. The results of Elisa showed that artesunate could induce apoptosis of HBZY-1 cells in a dose-dependent manner, and the results showed that: compared with normal control group, artesunate could induce apoptosis of HBZY-1 cells in a dose-dependent manner. The content of TNF- 伪 and IL-6 in the supernatant of high glucose culture model group was significantly higher than that in the high glucose culture model group, and the content of TNF- 伪 IL-6 in the supernatant of each artesunate drug group was lower than that of enalapril group, and the effect of high dose artesunate group was more obvious than that of enalapril group. Conclusion: high glucose environment can promote the proliferation of HBZY-1 cells and the secretion of TNF- 伪 IL-6, and induce inflammatory reaction. Artesunate could significantly reverse the proliferation and apoptosis of GMCs stimulated by high glucose, and inhibit the secretion of TNF- 伪-伪 IL-6 by HBZY-1 cells under high glucose. Artesunate can be used as an effective drug to prevent and treat diabetic nephropathy and reduce the damage to tissues and organs caused by high glucose environment.
【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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