高亲和力IgE受体FcεR1与非酒精性脂肪肝病关系的实验研究

发布时间：2018-06-29 06:01

本文选题：非酒精性脂肪肝病 + 肝功能　；参考：《山东大学》2017年硕士论文

【摘要】：[背景/目的]非酒精性脂肪肝病(nonalcoholic fatty liver disease,NAFLD)是常见以脂质堆积、酶学指标持续异常为特点的临床病理表现,是慢性肝脏疾病中的一个常见原因,是世界上一个重大的问题。调查结果显示,肥胖促进NAFLD病情加重。免疫球蛋白E(IgE)是介导I型变态反应的一种免疫球蛋白,高亲和力IgE受体(FcεRl)是IgE的受体之一。Yun-Jung Lee等人做了一项研究,检测随机抽取的50名肥胖妇女的血浆IgE,体质量和体重指数(BMI),粗略估计脂肪的含量,对此数据进行分析,发现血浆IgE水平随其他三项指标的升高而降低。并且做了相关动物实验,发现高脂饮食(HFD)诱导的FcεR1-/-肥胖小鼠进食量增加和能量代谢降低,导致体质量增加。本研究旨在通过HFD诱导小鼠NAFLD模型,探讨FcdR1在NAFLD发生发展中的作用。[方法]1.NAFLD模型的建立及指标检测:随机选取野生型C57BL/6J和IgE受体敲除FcεR1R/-、鼠各10只,分为野生型组(WT)和基因敲除组(FcεR1R-/-),均给予HFD喂养,在每周一定的时间称取其体质量。喂养12周后,小鼠禁食12h,称取体质量后处死取材。取全血,离心后取血浆,用生化检测仪检测血浆中谷丙转氨酶(ALT)、谷草转氨酶(AST)、甘油三酯(TG)、总胆固醇(TC)等指标。马上快速剪断肝血管并取出肝脏,称取重量后分别制成组织石蜡切片和冰冻切片,分别用于HE、masson和油红O染色,剩余部分置于-80℃C搁置,用于检测肝组织中TG、Ⅰ型胶原(collagenⅠ)和Ⅲ型胶原(collagenⅢ)等指标。2.胰岛素耐受实验(ITT):HFD喂养小鼠第10周时,禁食6h,腹部打入胰岛素(0.66IU/kg),打的同时开始计时,分别于0、30、60、90、120、150min用血糖针扎尾取血,用血糖仪检测血糖值,根据系列数据作ITT曲线,观察小鼠对胰岛素的耐受情况。3.葡萄糖耐受实验(GTT):HFD喂养小鼠第11周时,禁食12h,腹部打入葡萄糖(2g/kg),打的同时开始计时,分别于0、30、60、90、120、150min用血糖针扎尾取血,检测血糖值,根据系列数据作GTT曲线,观察小鼠对葡萄糖的控制情况。[结果]1.体质量和肝脏体重比的比较结果:与WT小鼠相比,FcεR1-/-小鼠体质量增长速度和肝脏体重比均明显增加。2.肝功能和肝组织纤维化的比较结果:与WT小鼠相比,FcεR1-/-、鼠血浆ALT和AST水平升高,肝组织collagenⅢ含量增加,collagenⅠ含量无明显变化,肝组织HE染色提示小鼠肝细胞肿胀及空泡样变较重,肝组织masson染色提示肝脏中纤维生成和堆积量增多,且阳性面积比增加。3.脂质代谢的比较结果:与WT小鼠相比,FcεR1-/-小鼠血浆TG和TC水平显著升高,小鼠肝组织TG堆积量增加,油红O染色提示肝脏脂质堆积程度较重,且阳性面积比增加。4.ITT和GTT的比较结果:WT和FcεR1-/-两组小鼠的ITT和GTT实验结果无明显差异,但是均对胰岛素敏感性下降,对葡萄糖控制较差。[结论]敲除FcεR1后,HFD诱导的NAFLD小鼠体质量升高,脂质代谢杂乱,NAFLD程度加重,而胰岛素抵抗(IR)无明显改变,提示FcεR1通过调节脂质代谢,不同程度的干预了小鼠NAFLD的进展,而此过程与IR无关。
[Abstract]:[background / purpose] nonalcoholic fatty liver disease (NAFLD) is a common clinical pathological manifestation characterized by lipid accumulation and persistent abnormalities of enzymology. It is a common cause of chronic liver disease. It is a major problem in the world. The results of investigation show that obesity promotes the aggravation of NAFLD, immune ball. Protein E (IgE) is an immunoglobulin which mediates I type allergy. The high affinity IgE receptor (Fc e Rl) is one of the IgE receptor.Yun-Jung Lee et al. A study was conducted to detect the plasma IgE, body mass and body mass index (BMI) of 50 randomly selected obese women. The content of fat was roughly estimated. The data were analyzed and plasma IgE was found. The level decreased with the increase of the other three indicators. And in the related animal experiments, we found that high fat diet (HFD) induced Fc epsilon R1-/- obese mice had increased food intake and reduced energy metabolism, resulting in increased body mass. The purpose of this study was to explore the role of FcdR1 in the development of NAFLD in mice by HFD induced NAFLD model. [method]1.NAFLD model]. Model establishment and index detection: random selection of wild type C57BL/6J and IgE receptor knockout Fc epsilon R1R/-, 10 rats each, divided into wild type group (WT) and gene knockout group (Fc epsilon R1R-/-), were given HFD feeding, and were given the body mass at the time fixed every Monday. After feeding 12 weeks, the mice were fasted 12h after being weighed. Take the whole blood, take the blood after centrifugation and take blood. Plasma, the serum ALT, AST, TG, and total cholesterol (TC) were detected by the biochemical analyzer. The liver vessels were quickly cut off and removed from the liver. After the weight, the tissue paraffin section and frozen section were made respectively, and respectively used for HE, Masson and oil red O staining, and the remaining part was placed in -80 and C shelved. TG, type I collagen (collagen I) and type III collagen (collagen III) were tested for.2. insulin tolerance test (ITT). After tenth weeks of HFD feeding mice, 6h was fasted, insulin (0.66IU/kg) was injected into the abdomen, and the timing was started at the same time. .3. glucose tolerance test (GTT) of mice's tolerance to insulin (GTT) was observed in a series of data. At eleventh weeks of HFD feeding mice, HFD was fed to 12h, the abdomen was injected into glucose (2g/kg), and the time was recorded at the same time. The blood glucose was measured by the tail of blood glucose needle in 0,30,60,90120150min, and the blood glucose was detected, and the GTT curve was observed according to the series of data. The mice were observed and observed. The control of glucose was compared with the results of]1. body mass and liver weight ratio: compared with WT mice, the growth rate of Fc epsilon R1-/- mice and the liver weight ratio significantly increased the results of.2. liver function and liver tissue fibrosis: compared with the WT mice, Fc e R1-/-, the increase of ALT and AST levels in the rat plasma, and the collagen III containing liver tissue There was no obvious change in the content of collagen I. The liver tissue HE staining showed that the liver cells were swelling and the vacuoles were heavier. Masson staining in the liver showed the increase of fibrous formation and accumulation in the liver, and the positive area was compared with the increase of.3. lipid metabolism: compared with the WT mice, the level of TG and TC in the plasma of Fc e R1-/- mice was significantly higher. The accumulation of TG in the rat liver was increased. The lipid accumulation in the liver was heavier with the oil red O staining, and the positive area was compared with the increase of.4.ITT and GTT. There was no significant difference between the ITT and GTT experimental results of WT and Fc epsilon two groups, but the sensitivity to insulin decreased and the control of grape sugar was poor. [Conclusion] after knocking Fc epsilon R1, HFD induction The body mass of LD mice increased, lipid metabolism was disorderly, the degree of NAFLD increased, and the insulin resistance (IR) had no obvious change. It suggested that Fc e R1 intervened the progress of NAFLD in mice by regulating lipid metabolism, and this process was not related to IR.
【学位授予单位】：山东大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R575.5

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