兰州大学机构库 >学院待认领
黄芪多糖对高糖状态下人肾小球系膜细胞IL-8和IL-10表达的影响
Alternative TitleThe effects of astragalus polysaccharide on expression of IL-8 and IL-10 secreted by human mesangial cells under high glucose
张秀德
Thesis Advisor陈慧
2013-05-21
Degree Grantor兰州大学
Place of Conferral兰州
Degree Name硕士
Keyword黄芪多糖 高糖状态 人肾小球系膜细胞 白介素-8 白介素-10
Abstract目的 观察黄芪多糖(APS)对高糖状态下人肾小球系膜细胞(HMCs)体外增殖作用,研究其对HMCs细胞中IL-8和IL-10表达的影响,探讨炎症反应因子在糖尿病肾病的作用。 方法 通过对HMCs细胞体外培养,选取对数生长期细胞进实验分组,分为对照组(NG组,5.5mmol/L D-葡萄糖),甘露醇高渗组(M组,24.5mmol/L甘露醇),高糖组(HG组,30 mmol/L D-葡萄糖),高糖+不同浓度黄芪多糖组(APS组),浓度分别为50ug/ml(APS50)、100ug/ml(APS100)、200ug/ml(APS200)、400ug/ml(APS400)、800ug/ml(APS800)及1600ug/ml(APS1600)。将HMCs细胞培养于30mmol/L D-葡萄糖的高糖环境中,以分组浓度APS分别干预12h、24h和48h后,MTT比色法检测各组细胞增殖程度,筛选适宜药物浓度;RT-PCR法检测各组HMCs细胞IL-8及IL-10 mRNA表达水平;免疫组织化学法测定各组HMCs细胞IL-8和IL-10蛋白表达水平。 结果 1.与NG组比较,高糖可明显促进HMCs细胞增殖(P<0.05),呈时间依赖性;与HG组比较,各浓度APS可明显抑制高糖状态下HMCs细胞增殖,具有统计学意义(P<0.05),且呈时间及剂量依赖性。2.与NG组比较,高糖可显著促进HMCs细胞IL-8和IL-10 mRNA及蛋白的表达(P<0.05);与HG组比较,APS可明显抑制高糖状态下HMCs细胞IL-8mRNA及蛋白表达,促进IL-10 mRNA及蛋白表达(P<0.05),且呈时间及剂量依赖性。 结论 黄芪多糖可抑制高糖状态下HMCs细胞过度增殖,下调高糖状态下HMCs细胞IL-8mRNA及蛋白的表达,上调IL-10mRNA及蛋白的表达,可能通过减轻炎症反应对DN肾脏起到保护作用。
Other AbstractObjective: To observe the effects of astragalus polysaccharides (APS) on the proliferation of the human mesangial cells (HMCs) in vitro and on the expression of Interleukin-8(IL-8) and Interleukin-10 (IL-10) in HMCs under high glucose state. Meanwhile, we explored the role of inflammation factors in diabetic nephropathy. Methods: Human mesangial cells were cultured in vitro, and selected the logarithmic growth phase cells into experiment. Then the cells were divided into control group (NG,5.5mmol/L D-glucose ) , mannitol group(M, 24.5mmol/L mannitol), high glucose group(HG, 30mmol/L D-glucose) , high glucose(30mmol/l) + APS of different concentrations group (APS) , the concentrations were 50ug/ml (APS50), 100ug/ml (APS100), 200ug/ml (APS200), 400ug/ml(APS400), 800ug/ml (APS800) and 1600ug/ml (APS1600 ). HMCs cells were cultured in 30 mmol/L glucose state, and intervented with astragalus polysaccharide of different concentrations for 12h, 24h and 48h. The proliferation of mesangial cells was detected at 12, 24 and 48h with the MTT to select the suitable drug concentration. The mRNA level of IL-8 and IL-10 and the protein themselves secreted by the mesangial cells under high glucose state were examined by RT-PCR and immunohistochemistry at 12, 24 and 48h. Results: 1. Compared with the NG group, high glucose state could significantly promote the proliferation of HMCs cell (P <0.05), and in a time-dependent manner. And compared with the HG group, APS of different concentrations significantly inhibited the proliferation of HMCs cell under high glucose state (P <0.05), and in a time-and dose-dependent manner. 2. Compared with the NG group, the condition of high glucose could sharply increase the expression of IL-8 and of IL-10 in HMCs cells (P < 0.05); Compared with the HG group, APS was able to reduce the expression of IL-8, and increase the expression of IL-10 in HMCs cells under high glucose significantly (P <0.05), and also in a time-and dose-dependent manner. Conclusion: The astragalus polysaccharides can inhibit the proliferation of HMCs under high glucose state. It may achieve the protective effect on DN by down-regulating the expression of the proinflammatory cytokines of IL-8 and up-regulating the expression of anti-inflammatory cytokine of IL-10.
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Language中文
Document Type学位论文
Identifierhttps://ir.lzu.edu.cn/handle/262010/223311
Collection学院待认领
Affiliation临床医学院
Recommended Citation
GB/T 7714
张秀德. 黄芪多糖对高糖状态下人肾小球系膜细胞IL-8和IL-10表达的影响[D]. 兰州. 兰州大学,2013.
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