细胞色素P4502E1调控酒精与高脂协同所致脂肪肝炎的小鼠病理模型构建

发布时间：2018-07-03 07:59

本文选题：细胞色素P450_2E_1 + 脂肪性肝炎　；参考：《川北医学院》2017年硕士论文

【摘要】：目的:(1)建立细胞色素P450_2E_1(CYP2E1)基因敲除鼠的肥胖和酒精协同增效性脂肪性肝炎动物模型。(2)探讨CYP2E1在酒精与高脂协同增效性脂肪性肝炎中的作用。方法:首先繁殖饲养CYP2E1基因敲除小鼠,并通过鼠尾基因鉴定筛选纯合子。然后分别将7周龄雄性CYP2E1基因敲除鼠和野生型小鼠同时分为4个处理组:1)正常饲料(对照)组:胃造瘘术后,通过微量注射泵每日以2μl/min的流速,580kcal/kg/d的液体常规饲料,恒速恒量灌注小鼠;2)酒精组:胃造瘘术后,通过微量注射泵每日以2ul/min的流速,580kcal/kg/d的液体常规饲料及15g/kg/d的酒精,恒速恒量灌注小鼠;3)高脂高胆固醇组:胃造瘘术后,通过微量注射泵每日以2μl/min的流速,780kcal/kg/d的液体高脂饲料(含0.21%的胆固醇)恒速恒量灌注小鼠;4)酒精协同高脂高胆固醇组:胃造瘘术后,通过微量注射泵每日以2μl/min的流速,780kcal/kg/d的液体高脂饲料(含0.21%的胆固醇)及15g/kg/d的酒精,恒速恒量灌注小鼠。同时饲喂2周后,取肝脏冰冻切片样本进行油红O、常规HE染色分析。结果:提取肝脏组织进行病理学分析,发现肝脏细胞出现气球样变,偶见点状坏死;油红O染色可见大量脂肪滴。同时通过Real-Time q PCR检测肝脏组织,发现酒精和高脂都能促进CYP2E1表达量增加,促进炎症因子表达,表明肥胖酒精协同增效性脂肪肝造模成功。结论:(1)成功构建小鼠高脂与酒精协同作用下的肝炎模型。(2)发现CYP2E1在高脂、酒精、及高脂酒精共同作用下都有介导炎症的作用。
[Abstract]:Objective: (1) to establish an animal model of obese and alcoholic synergistic fatty hepatitis in mice with cytochrome P4502E-1 (CYP2E1) knockout. (2) to investigate the role of CYP2E1 in synergistic fatty hepatitis between alcohol and hyperlipidemia. Methods: CYP2E1 knockout mice were bred and homozygotes were screened by mouse tail gene identification. Then male CYP2E1 knockout mice and wild-type mice were divided into four treatment groups: control group (control group) and male CYP2E1 knockout mouse group (control group). After gastrostomy, the rats were given a daily liquid diet of 580kcal / kg / d at a flow rate of 2 渭 l/min per day. After gastrostomy, the rats were perfused with constant speed and constant dose of alcohol at the flow rate of 580kcal / kg / d of liquid diet and alcohol of 15g/kg/d daily by microinjection pump. 3) the group of mice with high fat and high cholesterol was perfused with constant speed and constant quantity. After gastrostomy, the rats were treated by gastrostomy, and the rats were treated with high fat and high cholesterol after gastrostomy. After gastrostomy, mice were fed with liquid high-fat diet (containing 0.21% cholesterol) at a flow rate of 780kcal / kg / d at a flow rate of 2 渭 l/min per day by a microinjection pump. The mice were perfused with alcohol and hyperlipidemia and hypercholesterolemia group. Mice were infused with microinjection pump at a flow rate of 780kcal / kg / d of liquid high-fat diet (containing 0.21% cholesterol) and 15g/kg/d alcohol daily. After feeding for 2 weeks, the frozen liver sections were collected and analyzed by routine HE staining. Results: pathological analysis of liver tissues showed that there were balloon degeneration and occasional necrosis in liver cells, and a large number of fat droplets were found in oil red O staining. At the same time, the liver tissue was detected by Real-Time Q PCR. It was found that both alcohol and hyperlipidemia could increase the expression of CYP2E1 and promote the expression of inflammatory factors, which indicated that the model of fatty liver induced by obesity and alcohol synergism was successful. Conclusion: (1) the mouse model of hepatitis induced by hyperlipidemia and alcohol was successfully constructed. (2) it was found that CYP2E1 could mediate inflammation under the combined action of high fat, alcohol and high fat alcohol.
【学位授予单位】：川北医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R575.5

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