茶多酚、Vitamin C和Vitamin E对三聚氰胺及其与三聚氰酸联合攻毒小鼠免疫功能的调节作用

发布时间：2018-03-25 20:11

  本文选题：三聚氰胺　切入点：三聚氰酸　出处：《沈阳农业大学》2017年硕士论文

【摘要】：三聚氰胺(Melamine)是典型的三嗪类有机杂环化合物,它作为一种化工原料,被广泛应用于塑料、化肥等工业生产。三聚氰酸(Cyanuric acid)是三聚氰胺经连续脱氨基反应生成的水解产物。目前,国内外对于该类物质的研究主要集中在对机体器官、组织所造成的肾毒性、肝毒性、生殖毒性和遗传毒性等方面,关于它们对机体免疫系统的毒性作用研究还较为有限,能够调节该类物质毒性的天然解毒剂的研究是目前研究热点。本论文旨在研究三聚氰胺及其与三聚氰酸联合对雄性小鼠免疫功能的影响,同时探索了茶多酚、Vitamin C和Vitamin E作为天然解毒剂对该类物质所起的调节作用。试验一茶多酚、Vitamin C和Vitamin E对三聚氰胺及其与三聚氰酸联合攻毒小鼠B细胞的调节作用研究方法:将56只雄性昆明小鼠随机分为三聚氰胺组(30mg.kg-1)、三聚氰胺+茶多酚组(30+400mg.kg-1)、三聚氰胺+Vitamin C 和 Vitamin E 组(30+215/286mg.kg-1)、三聚氰胺+三聚氰酸联合组(15+15mg.kg-1)、联合+茶多酚组(15+15+400mg.kg-1)、联合+Vitamin C和Vitamin E组(15+15+215/286mg.kg-1)及空白对照组7个试验组。小鼠隔天灌胃28d,腹腔注射1%绵羊红细胞用于激发机体免疫反应。眼球采血和颈椎脱臼处死小鼠,采用全自动血液分析仪和流式细胞仪检测小鼠外周血和浆细胞表面标志CD138+的表达率。无菌摘取脾脏,RT-PCR、Western Blot法检测Blimp-1基因和蛋白的相对表达量。结果表明:①与对照组相比,联合组、联合+茶多酚及联合+VitaminC和VitaminE组的白细胞的数量均显著升高(P0.05),而联合+茶多酚组小鼠外周血中白细胞的数量显著低于联合组(P0.05)。②从小鼠淋巴细胞的数量上看,联合组和联合+VitaminC和Vitamin E组与对照组比数值显著增加(P0.05),而联合+茶多酚组未出现显著性差异(P0.05)。③三聚氰胺组和联合组CD138+的表达率减少(P0.01),而其它试验组未呈现出显著性变化(P0.05)。④从小鼠脾脏内Blimp-1蛋白的相对表达量来看,联合攻毒组与空白对照组小鼠相比极显著下降(P0.01),而其它试验组无显著性差异(P0.05);各试验组与空白对照组比,Blimp-1 mRNA相对表达量并未表现出显著性的变化(P0.05)。说明该类物质能通过降低Blimp-1蛋白的表达量,使B淋巴细胞的成熟和分化受阻,而茶多酚、Vitamin C和Vitamin E可以在一定程度上调节这种抑制作用。试验二茶多酸、Vitamin C和Vitamin E对三聚氰胺及其与三聚氰酸联合攻毒小鼠T细胞的调节作用研究方法:试验动物分组及处理同试验一,流式细胞仪检测小鼠外周血中T细胞表面标志 CD3+、CD4+、CD8+,及小鼠 CTL 细胞表面标志 CD8a+CD44hiCD62Llow。RT-PCR、Western Blot检测Notch-1基因和蛋白的相对表达情况。结果表明:①联合攻毒组小鼠CD4+/CD8+的比值与空白对照组相比显著降低(P0.05),而联合+茶多酚组、联合+Vitamin C和Vitamin E组小鼠CD4+/CD8+的比值变化不显著(P0.05),CD4+与CD8+的比值降低意味着机体的免疫功能可能在一定程度上受到了抑制,而使用了茶多酚、VitaminC和VitaminE后比值得到恢复。②各试验组与空白对照组相比较小鼠外周血中的CTL细胞表达率并未出现显著性差异(P0.05),但均低于对照组。③联合攻毒组Notch-1基因的相对表达量较空白对照组极显著升高(P0.01),而联合+茶多酚组与联合组相比Notch-1 mRNA的相对表达量显著降低(P0.05),说明该类物质通过使Notch-1基因相对表达量的增多来调控机体内T细胞的分化和成熟。但三聚氰胺或者其与三聚氰酸联合作用于机体时,VitaminC和VitaminE会使Notch-1蛋白相对表达量显著升高(P0.05),其它试验组并没有发生显著变化(P0.05),这与mRNA的检测结果不一致,其原因还有待于进一步研究。以上研究说明茶多酚、Vitamin C和Vitamin E对该类物质的免疫毒性具有一定的调节作用。
[Abstract]:Melamine (Melamine) is a typical three triazine heterocyclic organic compounds, it is used as a chemical raw material, widely used in plastics, fertilizers and other industrial production. Cyanuric acid (Cyanuric acid) is the melamine continuous de hydrolysate amino reaction. At present, domestic and foreign research on this kind of material is mainly concentrated on the body organs, tissue caused by kidney toxicity, liver toxicity, reproductive toxicity and genetic toxicity, about them on immune toxicity study is still limited, can adjust the toxic substance natural antidote research is the current research focus. This paper aims to study the melamine and cyanuric acid and its influence on the immune function of male mice, and explore the role of tea polyphenols, Vitamin C and Vitamin E as a natural antidote to this kind of material. The test of tea polyphenols, Vit Effect of Amin C and Vitamin E methods of melamine and cyanuric acid combined with cytotoxic attack B mice: 56 male Kunming mice were randomly divided into group (30mg.kg-1), melamine, melamine + tea polyphenol group (30+400mg.kg-1), +Vitamin C and Vitamin E group of melamine (30+215/286mg.kg-1), melamine + cyanuric acid group (15+15mg.kg-1 +), combined with tea polyphenols (15+15+400mg.kg-1), C and Vitamin combined with +Vitamin E group (15+15+215/286mg.kg-1) and blank control group 7 experimental groups. The mice were gavaged with 28d, abdominal cavity injection of 1% sheep red blood cells to stimulate immune response. The eye blood and the mice were killed by cervical dislocation by automatic blood analyzer and flow cytometry in peripheral blood and plasma cell surface marker CD138+ expression rate. Sterile removal of spleen, RT-PCR, detection of Blimp-1 Western by Blot gene and protein The relative expression level. The results showed that: compared with the control group, the combined group, the number of white blood cells + combined with tea polyphenols and the combination of +VitaminC and VitaminE group were significantly increased (P0.05), and combined with tea polyphenols + white cells in peripheral blood of mice was significantly lower than the number of combined group (P0.05) from the. The number of lymphocytes of mice at the combined group and the combination of +VitaminC and Vitamin in E group and control group increased significantly (P0.05), and combined with numerical + TP group did not show significant difference (P0.05). The expression of melamine group and combined group CD138+ (P0.01), and reduce the rate of other groups did not show up significant changes (P0.05). The Blimp-1 from the spleen of mice in the expression of the protein, combined with poison attack group and blank control group mice decreased significantly (P0.01), while there was no significant difference between the other experimental group (P0.05); the experimental group compared with blank control group, Blimp-1 M The relative expression of RNA did not show significant changes (P0.05). The material can reduce the expression of Blimp-1, B lymphocyte maturation and differentiation was blocked, and tea polyphenols, Vitamin C and Vitamin E can inhibit the regulation in a certain extent. Test two theapolyacid, regulation to study the effect of Vitamin C and Vitamin E methods of melamine and cyanuric acid combined with cytotoxic attack T mice: experimental animal groups and treatments were same, flow cytometry in mice T cell surface markers in peripheral blood CD3+, CD4+, CD8+, and mouse CTL cell surface marker CD8a+CD44hiCD62Llow.RT-PCR, the relative expression of Western Blot detection of Notch-1 gene and protein. The results showed that the combined ratio and blank mice challenged group CD4+/CD8+ compared with the control group decreased significantly (P0.05), and combined with tea polyphenols + group, combined with +Vitamin C and V The change of the ratio of itamin in E group CD4+/CD8+ was not significant (P0.05), the ratio of CD4+ and CD8+ mean lower immune function may to some extent inhibited, and the use of tea polyphenols, recovery ratio of VitaminC and VitaminE. The expression of the test group and blank control group compared with mouse peripheral CTL in the blood cell rate did not show significant difference (P0.05), but were lower than the control group. The combined inoculation group the expression of Notch-1 was significantly higher than the control group (P0.01), and the joint + TP group and combination group compared the relative expression of Notch-1 mRNA decreased significantly (P0.05) this kind of material that, by making the Notch-1 relative expression increased to differentiation and maturation of T cells in the body. But the regulation of melamine and cyanuric acid and its effect on the body, VitaminC and VitaminE can make Notch-1 protein. The expression increased significantly (P0.05), the other experimental group did not change significantly (P0.05), this inconsistency detection and mRNA results, the reason remains to be further studied. The above research shows that tea polyphenols, Vitamin C and Vitamin E has a certain effect on regulating the immune toxicity of the substances.

【学位授予单位】：沈阳农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S859.8
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本文编号：1664691