發(fā)布時(shí)間：2018-01-20 08:11

  本文關(guān)鍵詞： 馬里苷 雪菊多糖 胰島細(xì)胞 柚皮苷 人工抗原　出處：《新疆醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:1.探討兩色金雞菊黃酮成分馬里苷對(duì)胰島β細(xì)胞的保護(hù)作用及可能機(jī)制;2.探討雪菊多糖對(duì)胰島β細(xì)胞的增殖活性和胰島素分泌情況的影響;3.合成并鑒定馬里苷結(jié)構(gòu)類似物柚皮苷的人工完全抗原,為制備柚皮苷的單克隆抗體及建立相應(yīng)的免疫學(xué)分析方法敲除小分子柚皮苷奠定基礎(chǔ)。方法:1.運(yùn)用四甲基偶氮唑鹽比色法(MTT)檢測(cè)馬里苷對(duì)MIN6細(xì)胞增殖的影響;酶聯(lián)免疫法(ELISA)檢測(cè)胰島素分泌量;Western Blot方法檢測(cè)細(xì)胞中BAX,Bcl-2蛋白表達(dá)水平;2.運(yùn)用四甲基偶氮唑鹽比色法(MTT)檢測(cè)雪菊多糖對(duì)MIN6細(xì)胞增殖的影響;酶聯(lián)免疫法(ELISA)檢測(cè)胰島素分泌量;3.采用過碘酸鹽氧化法合成柚皮苷人工抗原;紫外光譜和薄層色譜法鑒定人工抗原偶聯(lián)是否成功。結(jié)果:1.與正常對(duì)照組比較,高糖高脂模型組MIN6細(xì)胞存活率明顯降低(P0.05),不同濃度馬里苷干預(yù)組MIN6細(xì)胞增殖均顯著增加(P0.05)。與正常對(duì)照組比較,高糖高脂模型組MIN6細(xì)胞胰島素分泌量明顯降低(P0.05);與高糖高脂模型組比較,馬里苷給藥組MIN6細(xì)胞胰島素分泌量明顯升高(P0.05)。Western印跡顯示,與正常對(duì)照組比較,BAX蛋白表達(dá)量模型組增多,給藥組隨濃度遞增依次減少;Bcl-2蛋白相對(duì)模型組減少,給藥組隨濃度增加依次增多(P0.05);2.與正常對(duì)照組比較,高糖高脂模型組細(xì)胞存活率明顯降低(P0.05),不同濃度雪菊多糖干預(yù)組MIN6細(xì)胞增殖均顯著增加(P0.05)。與正常對(duì)照組比較,高糖高脂模型組細(xì)胞胰島素分泌量明顯降低(P0.05);與高糖高脂模型組比較,給藥組細(xì)胞胰島素分泌量明顯升高(P0.05);3.紫外光譜圖譜顯示人工抗原偶聯(lián)成功;薄層色譜法顯示人工抗原偶聯(lián)成功。結(jié)論:1.馬里苷對(duì)高糖高脂造成的胰島細(xì)胞損傷有保護(hù)作用,其可能機(jī)制可能與Bcl蛋白家族有關(guān);2.雪菊多糖可以逆轉(zhuǎn)糖脂毒性對(duì)胰島細(xì)胞的損傷;3.成功合成了柚皮苷人工抗原,可用于下一步柚皮苷單克隆抗體制備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.
【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R285

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