两种两色金鸡菊活性成分对胰岛细胞保护作用及小分子敲除初步探索

发布时间：2018-01-20 08:11

本文关键词： 马里苷雪菊多糖胰岛细胞柚皮苷人工抗原　出处：《新疆医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:1.探讨两色金鸡菊黄酮成分马里苷对胰岛β细胞的保护作用及可能机制;2.探讨雪菊多糖对胰岛β细胞的增殖活性和胰岛素分泌情况的影响;3.合成并鉴定马里苷结构类似物柚皮苷的人工完全抗原,为制备柚皮苷的单克隆抗体及建立相应的免疫学分析方法敲除小分子柚皮苷奠定基础。方法:1.运用四甲基偶氮唑盐比色法(MTT)检测马里苷对MIN6细胞增殖的影响;酶联免疫法(ELISA)检测胰岛素分泌量;Western Blot方法检测细胞中BAX,Bcl-2蛋白表达水平;2.运用四甲基偶氮唑盐比色法(MTT)检测雪菊多糖对MIN6细胞增殖的影响;酶联免疫法(ELISA)检测胰岛素分泌量;3.采用过碘酸盐氧化法合成柚皮苷人工抗原;紫外光谱和薄层色谱法鉴定人工抗原偶联是否成功。结果:1.与正常对照组比较,高糖高脂模型组MIN6细胞存活率明显降低(P0.05),不同浓度马里苷干预组MIN6细胞增殖均显著增加(P0.05)。与正常对照组比较,高糖高脂模型组MIN6细胞胰岛素分泌量明显降低(P0.05);与高糖高脂模型组比较,马里苷给药组MIN6细胞胰岛素分泌量明显升高(P0.05)。Western印迹显示,与正常对照组比较,BAX蛋白表达量模型组增多,给药组随浓度递增依次减少;Bcl-2蛋白相对模型组减少,给药组随浓度增加依次增多(P0.05);2.与正常对照组比较,高糖高脂模型组细胞存活率明显降低(P0.05),不同浓度雪菊多糖干预组MIN6细胞增殖均显著增加(P0.05)。与正常对照组比较,高糖高脂模型组细胞胰岛素分泌量明显降低(P0.05);与高糖高脂模型组比较,给药组细胞胰岛素分泌量明显升高(P0.05);3.紫外光谱图谱显示人工抗原偶联成功;薄层色谱法显示人工抗原偶联成功。结论:1.马里苷对高糖高脂造成的胰岛细胞损伤有保护作用,其可能机制可能与Bcl蛋白家族有关;2.雪菊多糖可以逆转糖脂毒性对胰岛细胞的损伤;3.成功合成了柚皮苷人工抗原,可用于下一步柚皮苷单克隆抗体制备及小分子敲除方法的建立。
[Abstract]:Objective: 1. To investigate the protective effect and possible mechanism of the flavonoid component of Chrysanthemum bicolor on islet 尾 cells. 2. To investigate the effects of snow chrysanthemum polysaccharides on the proliferation activity and insulin secretion of islet 尾 cells. 3. The artificial complete antigen of naringin was synthesized and identified. In order to prepare the monoclonal antibody of naringin and establish the corresponding immunological analysis method for knockout naringin. Methods: 1. Using tetramethylazolium colorimetric method (MTT). The effect of Marinin on proliferation of MIN6 cells was detected. Enzyme linked immunosorbent assay (Elisa) was used to detect insulin secretion. Western Blot method was used to detect the expression of Bax Bcl 2 protein in the cells. 2. The effect of polysaccharides on the proliferation of MIN6 cells was detected by tetramethylazolium colorimetric assay (MTT). Enzyme linked immunosorbent assay (Elisa) was used to detect insulin secretion. 3. Naringin artificial antigen was synthesized by periodate oxidation method. UV spectra and TLC were used to identify the success of artificial antigen coupling. Results 1. Compared with the normal control group, the survival rate of MIN6 cells in the high glucose and high fat group was significantly lower than that in the control group (P 0.05). The proliferation of MIN6 cells was significantly increased in the different concentration of Marinoside intervention group compared with the normal control group. The insulin secretion of MIN6 cells in high glucose and high fat group was significantly decreased (P 0.05). Compared with the model group of high glucose and high fat, the insulin secretion of MIN6 cells in the group treated with Marinside was significantly higher than that in the control group (P 0.05N. Western blotting), and compared with that in the normal control group. The expression of BAX protein increased in the model group and decreased with the increase of the concentration in the administration group. The Bcl-2 protein decreased in the model group, and increased with the increase of the concentration in the administration group (P 0.05). 2.Compared with the normal control group, the cell survival rate of the model group with high glucose and high fat decreased significantly (P 0.05). Compared with the normal control group, the insulin secretion of the high glucose and high fat model group decreased significantly (P 0.05). Compared with the model group of high glucose and high fat, the insulin secretion of the treated group was significantly higher than that of the control group. 3. The ultraviolet spectrum showed that the artificial antigen was successfully coupled. Thin-layer chromatography showed that artificial antigen was successfully coupled. Conclusion: 1. Marinoside has protective effect on islet cell damage induced by high glucose and high fat, and its possible mechanism may be related to Bcl protein family. 2.From polysaccharide can reverse the damage of islet cells caused by glycolipid toxicity; 3. Naringin artificial antigen was successfully synthesized, which can be used in the preparation of naringin monoclonal antibody and the establishment of small molecular knockout method.
【学位授予单位】：新疆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285

【参考文献】