王燕林,梁文妹,尹丹
WANG Yanling,LIANG Wenmei,YIN Dan

• 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:探讨大鼠摄入不同剂量的某品牌54度食用白酒后,不同时点大鼠肝细胞金属硫蛋白(MT)表达的变化。方法:115只正常雄性SD大鼠分为正常对照组(n=25)和低剂量组[灌胃白酒0.8 m L/(kg·d)]、中剂量组[灌胃白酒1.6 m L/(kg·d)]及高剂量组[灌胃白酒2.4 mL/(kg·d)]实验组,各组大鼠均给予正常饲料喂养,各白酒组还给予相应剂量的白酒灌胃;所有大鼠分别于灌胃第4、8、12、16及20周取肝脏组织,采用免疫组化SABC法、免疫蛋白印迹(Western Blot)检测肝细胞及肝组织匀浆中MT蛋白表达。结果:正常组及各实验组肝细胞MT免疫反应的阳性产物主要集中在胞浆,呈棕褐色细颗粒状;而肝小叶之间的结缔组织,血管、Kuffer细胞、胆管组织未见MT着色,所有阴性对照未见MT阳性产物;与正常对照组比较,第4、8、12、16及20周各实验组大鼠肝细胞内MT免疫反应阳性产物定位及免疫反应染色强度未见明显变化;MT、β-actin蛋白分别在15 k D、42 k D处显示出特异性条带;第4、8、12、16及20周各实验组MT蛋白表达水平与正常对照组相比,差异无统计学意义(P>0.05)。结论:在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MT表达水平无明显影响。
Objective: To investigate effects of a brand of edible liquor from Guizhou on metallothionein(MT) expression in rat hepatocytes at different time points after intake with different doses.Methods: One hundred and fifteen normal adult male SD rats were randomly divided into a normal control group(group C,n = 25) and several experimental groups including low dose group[group L,0. 8 m L/(kg·d) ],middle dose group[group M,1. 6 m L/(kg·d) ],and high dose group[groupH,2. 4 m L/(kg·d) ]. All of the rats were normally reared,and beside,rats in experimental groups were given liquor by gavage twice a day at corresponding doses. Rat liver tissue was taken at the end of the 4 th,8 th,12 th,16 th and 20 th weeks of the experiment,and the expression of MT level was detected by immunohistochemical SABC method and Western blotting. Results: MT immunohistochemical positive products existed within cytoplasm of hepatocyte; Western blotting results showed that the expression level of MT protein in the experimental groups at all of the time points did not differ from those of group C statistically significant(P > 0. 05). Conclusion: This liquor does not influence MT expression level in rat hepatocytes obviously at the does range and during the set time period,when administrated by gavage.

关键词(KeyWords)： 食用白酒;金属硫蛋白;肝细胞;免疫组织化学;蛋白免疫印迹
liquor;metallothionein;hepatocyte;immunohistochemistry;Western Blotting

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,尹丹
WANG Yanling,LIANG Wenmei,YIN Dan

DOI: 10.19367/j.cnki.1000-2707.2017.05.008

参考文献(References)：

• [1]Capaldo A,Gay F,Scudiero R,et al.Histological changes,apoptosis and metallothionein levels in Triturus carnifex(Amphibia,Urodela)exposed to environmental cadmium concentrations[J].Aquat Toxicol,2016(173):63-73.
• [2]Szrok S.Metallothioneins 1 and 2,but not 3,are regulated by nutritional status in rat white adipose tissue[J].Genes&Nutrition,2016(1):1-12.
• [3]Grasselli E,Compalati AD,Voci A,et al.Altered oxidative stress/antioxidant status in blood of alcoholic subjects is associated with alcoholic liver disease[J].Drug&Alcohol Dependence,2014(1):112-119.
• [4]O'Connor KS,Parnell G,Patrick E,et al.Hepatic metallothionein expression in chronic hepatitis C virus infection is IFNL3 genotype-dependent[J].Genes Immun,2014(2):88-94.
• [5]Cederbaum AI,Lu Y,Wu D.Role of oxidative stress in alcohol-induced liver injury[J].Arch Toxicol,2009(6):519-548.
• [6]刘爱东,谢岚,蒋慧,等.不同品质的食用白酒对大鼠肝脏形态及其肝功能的影响[J].酿酒科技,2014(1):125-128.
• [7]江源.2016年6月全国白酒产量116万千升[J].酿酒科技,2016(8):27.
• [8]Qu W,Waalkes MP.Metallothionein blocks oxidative DNA damage induced by acute inorganic arsenic exposure[J].Toxicol Appl Pharmacol,2015(3):267-274.
• [9]田鹏.水体中镉暴露对草鱼的氧化胁迫研究[D].重庆·西南大学,2013.
• [10]Nagamine T,Nakajima K.Significance of metallothionein expression in liver disease[J].Curr Pharm Biotechnol,2013(4):420-426.
• [11]叶兰,熊陶,黄能慧,等.高脂饲料和酒精诱导大鼠肝纤维化的实验研究[J].贵阳医学院学报,2003(2):152-154.
• [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)：