乙醇抑制蛋白酶体诱导酒精性肝病模型大鼠肝脏线粒体凋亡

发布时间：2018-01-12 15:17

本文关键词：乙醇抑制蛋白酶体诱导酒精性肝病模型大鼠肝脏线粒体凋亡　出处：《中华实用诊断与治疗杂志》2016年01期 　论文类型：期刊论文

【摘要】：目的探讨乙醇抑制蛋白酶体在酒精性肝病大鼠模型肝细胞线粒体凋亡中的作用。方法健康雄性Wistar大鼠40只,随机分为对照组、模型1组、模型2组、模型3组、模型4组各8只。模型组均采用乙醇灌胃构建酒精性肝病大鼠模型,对照组给予等量生理盐水。分别于造模6周(模型1组)、8周(模型2组)、10周(模型3组)、12周(对照组、模型4组)处死大鼠,取肝组织行组织病理学检查并应用电镜观察,采用流式细胞术检测肝细胞线粒体膜电位,采用RT-PCR法检测肝组织低分子多肽7(low molecular weight polypetide 7,LMP7)mRNA表达水平,采用Western blot检测肝组织LMP7蛋白表达水平。结果 (1)组织病理结果显示对照组肝小叶轮廓清晰,肝细胞围绕中央静脉呈正常索状排列,无变性、炎症及坏死;模型1组肝小叶中央区域少量脂肪变性;模型2组细胞内出现较多脂滴,肝细胞细胞核被推挤至细胞一侧;模型3组细胞内含大量脂滴,肝细胞肿大变圆;模型4组脂肪变进一步加重,可见不同程度肝细胞水样变性和炎细胞浸润及Mallory小体;(2)电镜结果显示模型组均成功构建酒精性肝病模型,线粒体在结构和形态上出现损伤,有凋亡表现;(3)模型组罗丹明123(rhodamine 123,Rho123)荧光强度[(239.46±12.72)%]明显低于对照组[(377.59±16.81)%](P0.05);(4)模型1、2、3、4组肝组织LMP7 mRNA相对表达量[0.51(0.31,0.87),0.49(0.15,0.79),0.08(0.03,0.14),0.02(0.01,0.03)]和LMP7蛋白表达水平(0.784±0.025、0.601±0.018、0.376±0.013、0.121±0.021)均明显低于对照组(1、0.997±0.031)(P0.05),模型1、2、3、4组LMP7mRNA相对表达量及LMP7蛋白表达水平依次降低,两两比较差异均有统计学意义(P0.05)。结论乙醇抑制蛋白酶体可能在酒精性肝病肝细胞线粒体凋亡中发挥重要作用。
[Abstract]:Objective to investigate the effect of ethanol inhibiting proteasome on apoptosis of hepatocyte mitochondria in rats with alcoholic liver disease. Methods Forty healthy male Wistar rats were randomly divided into control group and model group 1. Model 2 group, model 3 group, model 4 group 8 rats in each group. The model group were all treated with ethanol to establish the model of alcoholic liver disease, the control group was given the same amount of normal saline, and the model was made for 6 weeks (model group 1). The rats were killed at 8 weeks (model group 2) for 10 weeks (control group, model group 4) for 12 weeks (model group 3). Histopathological examination of liver tissue was performed and observed by electron microscope. Mitochondrial membrane potential of hepatocytes was measured by flow cytometry. RT-PCR method was used to detect the expression of low molecular polypeptide 7 molecular weight polypetide 7 mRNAs in liver tissue. Western blot was used to detect the expression of LMP7 protein in liver tissue. The liver cells were arranged in normal cord-like form around the central vein, without degeneration, inflammation and necrosis. Model 1 had a small amount of steatosis in the central region of hepatic lobule. In model 2, more lipid droplets appeared in the cells, and the nuclei of the hepatocytes were pushed to one side of the cells. Model group 3 cells contain a large number of lipid droplets, hepatocyte swelling round; In model group 4, fatty degeneration was further aggravated, with different degrees of hepatocyte hydrophilic degeneration, inflammatory cell infiltration and Mallory corpuscles. The results of electron microscope showed that the alcoholic liver disease model was successfully constructed in the model group. The mitochondria were damaged in structure and morphology and showed apoptosis. Rho123 Rho123) fluorescence intensity. [239.46 卤12.72%] was significantly lower than that in the control group. [The relative expression of LMP7 mRNA in the liver tissue of the model group 1, 2, 2, 3, 5 (1). [0.51U 0.31U 0.87m 0.49m 0.150.79A 0.08U 0.03U 0.140.2kW 0.02ng01. The main results are as follows: (1) 0. 51U 0. 31U 0. 87m. The expression level of LMP7 protein was 0.784 卤0.025, 0.601 卤0.018, 0.376 卤0.013. 0.121 卤0.021) was significantly lower than that in control group (0.997 卤0.031) (P 0.05N). The relative expression of LMP7mRNA and the expression level of LMP7 protein decreased in turn in 4 groups. Conclusion ethanol inhibition of proteasome may play an important role in apoptosis of hepatocyte mitochondria in alcoholic liver disease.
【作者单位】：东莞东华医院消化内科;山东大学齐鲁医院消化内科;
【基金】：山东省医药卫生科研重点项目(2007Hz056)
【分类号】：R575;R-332
【正文快照】： 流行病学调查结果[1]显示,酒精性肝病在我国的发病率和病死率呈上升趋势,已成为仅次于病毒性肝炎的第2大肝病。长期摄入乙醇造成氧化蛋白生成增多是肝细胞损伤的重要原因。凋亡与凋亡细胞的清除是保证肝功能正常的关键因素。蛋白酶体可降解细胞内大部分蛋白,通过降解诱导凋亡

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