本文關(guān)鍵詞：乙醇抑制蛋白酶體誘導(dǎo)酒精性肝病模型大鼠肝臟線粒體凋亡　出處：《中華實(shí)用診斷與治療雜志》2016年01期 　論文類型：期刊論文

  更多相關(guān)文章： 酒精性肝病 線粒體 凋亡 蛋白酶體 蛋白酶體糜蛋白酶樣活性亞基 大鼠

【摘要】：目的探討乙醇抑制蛋白酶體在酒精性肝病大鼠模型肝細(xì)胞線粒體凋亡中的作用。方法健康雄性Wistar大鼠40只,隨機(jī)分為對照組、模型1組、模型2組、模型3組、模型4組各8只。模型組均采用乙醇灌胃構(gòu)建酒精性肝病大鼠模型,對照組給予等量生理鹽水。分別于造模6周(模型1組)、8周(模型2組)、10周(模型3組)、12周(對照組、模型4組)處死大鼠,取肝組織行組織病理學(xué)檢查并應(yīng)用電鏡觀察,采用流式細(xì)胞術(shù)檢測肝細(xì)胞線粒體膜電位,采用RT-PCR法檢測肝組織低分子多肽7(low molecular weight polypetide 7,LMP7)mRNA表達(dá)水平,采用Western blot檢測肝組織LMP7蛋白表達(dá)水平。結(jié)果 (1)組織病理結(jié)果顯示對照組肝小葉輪廓清晰,肝細(xì)胞圍繞中央靜脈呈正常索狀排列,無變性、炎癥及壞死;模型1組肝小葉中央?yún)^(qū)域少量脂肪變性;模型2組細(xì)胞內(nèi)出現(xiàn)較多脂滴,肝細(xì)胞細(xì)胞核被推擠至細(xì)胞一側(cè);模型3組細(xì)胞內(nèi)含大量脂滴,肝細(xì)胞腫大變圓;模型4組脂肪變進(jìn)一步加重,可見不同程度肝細(xì)胞水樣變性和炎細(xì)胞浸潤及Mallory小體;(2)電鏡結(jié)果顯示模型組均成功構(gòu)建酒精性肝病模型,線粒體在結(jié)構(gòu)和形態(tài)上出現(xiàn)損傷,有凋亡表現(xiàn);(3)模型組羅丹明123(rhodamine 123,Rho123)熒光強(qiáng)度[(239.46±12.72)%]明顯低于對照組[(377.59±16.81)%](P0.05);(4)模型1、2、3、4組肝組織LMP7 mRNA相對表達(dá)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蛋白表達(dá)水平(0.784±0.025、0.601±0.018、0.376±0.013、0.121±0.021)均明顯低于對照組(1、0.997±0.031)(P0.05),模型1、2、3、4組LMP7mRNA相對表達(dá)量及LMP7蛋白表達(dá)水平依次降低,兩兩比較差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論乙醇抑制蛋白酶體可能在酒精性肝病肝細(xì)胞線粒體凋亡中發(fā)揮重要作用。
[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 鹵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 鹵0.025, 0.601 鹵0.018, 0.376 鹵0.013. 0.121 鹵0.021) was significantly lower than that in control group (0.997 鹵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.
【作者單位】： 東莞東華醫(yī)院消化內(nèi)科;山東大學(xué)齊魯醫(yī)院消化內(nèi)科;
【基金】：山東省醫(yī)藥衛(wèi)生科研重點(diǎn)項(xiàng)目(2007Hz056)
【分類號】：R575;R-332
【正文快照】： 流行病學(xué)調(diào)查結(jié)果[1]顯示,酒精性肝病在我國的發(fā)病率和病死率呈上升趨勢,已成為僅次于病毒性肝炎的第2大肝病。長期攝入乙醇造成氧化蛋白生成增多是肝細(xì)胞損傷的重要原因。凋亡與凋亡細(xì)胞的清除是保證肝功能正常的關(guān)鍵因素。蛋白酶體可降解細(xì)胞內(nèi)大部分蛋白,通過降解誘導(dǎo)凋亡

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