王燕林,梁文妹,李容,陈兴强
WANG Yanling,LIANG Wenmei,LI Rongrong,CHEN Xingqiang

• 1:贵州医科大学
• 2:贵州医科大学组织学与胚胎学教研室

摘要(Abstract)：

目的:探讨摄入某品牌54度食用白酒后,不同时长及剂量下,对大鼠肝细胞肿瘤坏鲜红因子(TNF-α)表达及凋亡的影响。方法:将115只正常雄性SD大鼠随机分为正常对照组(n=25)及实验组(n=90),实验组分为低剂量组[0.8 m L/(kg·d)]、中剂量组[1.6 m L/(kg·d)]、高剂量组[2.4 m L/(kg·d)];实验组大鼠每日上午11点及下午5点用该白酒各灌胃1次,正常组不予处理,分别于第4、8、12、16和20周末时取肝脏组织,采用免疫组化SABC法检测TNF-α表达,TUNEL法检测各组大鼠肝细胞凋亡、图像分析及形态计量法检测大鼠肝细胞TNF-α平均光密度值及凋亡指数(AI)。结果:正常对照组及各实验组肝细胞中,TNF-α免疫反应的阳性产物主要集中于胞浆,TUNEL检测肝细胞凋亡阳性产物存在于肝细胞核内,与正常对照组比较,第4周、8周、12周、16周及20周末时各实验组肝细胞内TNF-α免疫反应阳性产物位置及染色强度未见明显变化,AI也无明显改变,差异无统计学意义(P>0.05)。结论:本实验设定的剂量和时程内,用该白酒灌胃后,对大鼠肝细胞TNF-α表达水平及AI未见明显影响。
Objective: To investigate the effect of the expression of TNF-α and the apoptosis of rat hepatocytes after intake certain 54° edible liquor at different doses and schedules. Methods: 115 normal adult male SD rats were randomly divided into control group( n = 25) and an experimental group( n= 90). Each experimental group was further divided into low dose group[0. 8 m L/( kg·d) ],middle dose group[1. 6 m L/( kg·d) ],high dose group[2. 4 m L/( kg·d) ]. The experimental groups were fed by gavage twice daily( 11 ∶ 00 AM an 5 ∶ 00 PM) with the liquor,the control group received no treatment. At the end of the 4th,8th,12 th,16th and 20 th week,liver tissue was taken. The expression of TNF-α was detected by immunohistochemical SABC method; The apoptosis of hepatocyte was detected by terminal deoxynucleotidyl transferase mediated nick end labeling( TUNEL). The mean optical density of TNF-α and apoptosis index in rat hepatocytes were detected by image analysis and morphometry. Results: TNF-α immunohistochemical positive products existed in cytoplasm of hepatocyte;TUNEL test indicated hepatocyte apoptosis positive product exists in the nucleus; immunoreaction positive product location of hepatocyte TNF-α and the staining intensity in the experimental groups at the end of 4th,8th,12 th,16th,and 20 th week,has no obvious change,the same also applied to AI,differences were not statistically significantly( P > 0. 05). Conclusion: TNF-α expression level and metric measurement; alcoholic liver diseaseapoptotic index of hepatocyte in rat liver has no obvious change in this experiment at set doses and schedules after gavage with the liquor.

关键词(KeyWords)： 肿瘤坏死因子;肝细胞;凋亡;免疫组织化学;形态计量;酒精性肝病
tumor necrosis factor alpha;hepatocytes;apoptosis;immunohistochemistry;morphometric measurement;alcoholic liver disease

Abstract:

Keywords:

基金项目(Foundation): 贵州省教育厅“125”重大科技专项基金资助[黔教合重大专项字(2012)007]

作者(Author): 王燕林,梁文妹,李容,陈兴强
WANG Yanling,LIANG Wenmei,LI Rongrong,CHEN Xingqiang

DOI: 10.19367/j.cnki.1000-2707.2017.06.009

参考文献(References)：

• [1]Novo-Veleiro I,Alvela-Suarez L,Chamorro AJ,et al.Alcoholic liver disease and hepatitis C virus infection[J].World J Gastroenterol,2016(4):1411-1420.
• [2]Gemes K,Janszky I,Laugsand LE,et al.Alcohol consumption is associated with a lower incidence of acute myocardial infarction:results from a large prospective population-based study in Norway[J].J Intern Med,2016(4):365-375.
• [3]Xie W,Sun J,Zhang X,et al.Necrosis factor-alpha(TNF-alpha)response in human hepatoma Hep G2 cells treated with hepatotoxic agents[J].Pharmazie,2014(5):379-384.
• [4]Cederbaum AI,Lu Y,Wang X,et al.Synergistic toxic interactions between CYP2E1,LPS/TNFalpha,and JNK/p38 MAP kinase and their implications in alcohol-induced liver injury[J].Adv Exp Med Biol,2015(815):145-172.
• [5]Eum HA,Billiar TR.TNF/TNF receptor 1-mediated apoptosis in hepatocytes[J].Adv Exp Med Biol,2011(691):617-624.
• [6]Roerecke M,Nanau R,Rehm J,et al.Ethnicity matters:a systematic review and meta-analysis of the non-linear relationship between alcohol consumption and prevalence and incidence of hepatic steatosis[J].E Bio Medicine,2016(8):317-330.
• [7]Louvet A,Mathurin P.Alcoholic liver disease:mechanisms of injury and targeted treatment[J].Nat Rev Gastroenterol Hepatol,2015(4):231-242.
• [8]Potz BA,Lawandy IJ,Clements RT,et al.Alcohol modulates autophagy and apoptosis in pig liver tissue[J].JSurg Res,2016(1):154-162.
• [9]Filliol A,Piquet-Pellorce C,Raguenes-Nicol C,et al.RIPK1 protects hepatocytes from Kupffer cells-mediated TNF-induced apoptosis in mouse models of PAMP-induced hepatitis[J].J Hepatol,2017(1):1205.
• [10]Suyavaran A,Ramamurthy C,Mareeswaran R,et al.TNF-alpha suppression by glutathione preconditioning attenuates hepatic ischemia reperfusion injury in young and aged rats[J].Inflamm Res,2015(1):71-81.
• [11]尹丹,梁文妹,夏白娟,等.某食用白酒对大鼠血糖、胰岛素和胰高血糖素表达的影响[J].贵阳医学院学报,2015(12):1285-1289.
• [12]刘昊,梁文妹.贵州某白酒对大鼠睾丸间质细胞内类固醇激素急性调节蛋白表达及血清睾酮水平的影响[J].解剖学报,2016(3):386-390.

文章评论(Comment)：