口服HSP60诱导免疫耐受抗动脉粥样硬化模型的建立及评价

发布时间：2018-01-21 15:25

本文关键词： 口服免疫耐受动脉粥样硬化 HsP6O ApoE~(-/-)小鼠模型　出处：《吉林大学》2011年硕士论文　论文类型：学位论文

【摘要】：动脉粥样硬化(atherosclerosis, AS)所致的心、脑血管疾病已严重危害人类生命健康。随着社会的发展和各方面竞争压力的增加,动脉粥样硬化对人类健康的影响与日俱增。因此,建立理想的口服免疫耐受模型在抗动脉粥样硬化疾病的研究中有重要意义。本研究旨在通过构建动脉粥样硬化模型,检测口服小剂量热休克蛋白60(HSP60)干预对动脉粥样硬化模型的建立的影响。进一步分析口服HSP60与动脉粥样硬化的关系,为进一步研究口服免疫耐受在抗动脉粥样硬化的作用及机制及治疗心血管疾病提供新的靶点及理论依据。目的：口服HSP60诱导免疫耐受对动脉粥样硬化的影响及相关因素的评价。方法：健康雄性8周龄ApoE-/-小鼠,16只,随机分为高脂对照组、HSP60组。小鼠分别给予单纯蒸馏水和重组小鼠HSP60+生理盐水+长链甘油三酯隔天灌胃,共4次,末次灌胃7d后腹腔注射等剂量的HSP60,高脂饲养10周后进行实验。实验过程中每7-10天测定小鼠体重,实验结束后收集主动脉弓段组织制作病理组织切片,HE染色以观察组织学的变化及比较斑块面积大小。结果：(1)高脂饮食4周后高脂组小鼠出现嗜睡,饮食、饮水增多,一般活动减少。HSP60组小鼠行为表现及各种活动均正常。(2)高脂饮食4周后,高脂组与HSP60组体重增长比较具有统计学差异(P＜0.01),HSP60组体重增长慢于高脂组。(3)主动脉段HE染色：高脂组主动脉内斑块形成明显,主动脉管腔形成狭窄。HSP60组管腔内膜仅略不光滑,未见明显狭窄形成。(4)计算动脉粥样硬化斑块面积：病理图像分析仪计算斑块面积各组ApoE-／-小鼠主动脉粥样斑块面积,HSP60口服组主动脉粥样斑块面积(无斑块形成)明显小于高脂组斑块面积[(2202.89±524.25)μm2](P＜0.01)。结论：(1)、本实验通过高脂喂养ApoE-/-小鼠建立合理的动脉粥样硬化模型。(2)、应用HSP60具有明显的抗动脉粥样硬化作用。(3)、本实验所建立的动物模型可以用于口服免疫耐受抗动脉粥样硬化的研究,其为进一步阐明口服免疫耐受通过上调CD4+ CD25+调节性T细胞进而预防动脉粥样硬化及冠心病的发病与进展方面奠定了实验基础。
[Abstract]:Atherosclerosis caused by atherosclerotic disease (ASA) has been seriously harmful to human life and health. With the development of society and the increase of competition pressure in all aspects. The influence of atherosclerosis on human health is increasing. Therefore, it is important to establish an ideal oral immune tolerance model in the study of anti-atherosclerotic diseases. The purpose of this study was to construct an atherosclerosis model. To detect the effect of oral heat shock protein 60 (HSP60) on the establishment of atherosclerosis model, and to further analyze the relationship between oral HSP60 and atherosclerosis. To further study the role and mechanism of oral immune tolerance in anti-atherosclerosis and to provide a new target and theoretical basis for the treatment of cardiovascular disease. Objective: to evaluate the effect of oral HSP60 induced immune tolerance on atherosclerosis. Methods: sixteen healthy male 8-week-old ApoE-r-mice were randomly divided into hyperlipidemic control group. HSP60 group. Mice were given intraperitoneal injection of HSP60 of the same dose respectively with distilled water and recombinant HSP60 saline with long chain triglyceride for 4 times every other day. The body weight of mice was measured every 7-10 days after high-fat feeding for 10 weeks. After the experiment, the aortic arch tissues were collected to make pathological sections. HE staining was used to observe histological changes and to compare the size of plaques. Results (1) after 4 weeks of high-fat diet, mice in high-fat group appeared drowsiness, diet and drinking water increased. The general activity decreased. HSP60 group showed normal behavior and all kinds of activities. 2) after 4 weeks of high fat diet. There was significant difference in weight gain between high fat group and HSP60 group (P < 0.01). The weight gain of HSP60 group was slower than that of hyperlipidemia group. No significant stenoses formation was observed.) Atherosclerotic plaque area was calculated by pathological image analyzer. ApoE-r-a-mouse aortic atherosclerotic plaque area was calculated by pathological image analyzer. Atheromatous plaque area (no plaque formation) in HSP60 group was significantly smaller than that in high fat group. [2202.89 卤524.25 渭 m ~ 2] P < 0.01. Conclusion in this experiment, we established a reasonable atherosclerosis model by high-fat feeding ApoE-r-mice. HSP60 has obvious anti-atherosclerotic effect. The animal model established in this study can be used to study the anti-atherosclerosis of oral immune tolerance. It provides an experimental basis for further elucidation of oral immune tolerance in preventing atherosclerosis and coronary heart disease by upregulating CD4 CD25 regulatory T cells.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R-332;R543.5

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