当药和水飞蓟提取混合物对毒性物质诱导的脂肪肝大鼠肝损伤的防治作用  

Effects of the mixture of Swertia pseudochinensis Hara and Silybum marianum Gaertn extracts on CCl_4-induced liver injury in rats with non-alcoholic fatty liver disease

作　　者：毛志敏[1,2] 宋海燕[1,2] 杨丽丽[1,2] 柳涛[1,2] 励冬斐[1,2] 郑培永[1,2] 刘平 季光[1,2,3] 

机构地区：[1]上海中医药大学脾胃病研究所 [2]上海中医药大学龙华医院消化内科,上海200032 [3]上海高校中医内科学E-研究院,上海201203

出　　处：《中西医结合学报》2012年第2期193-199,共7页Journal of Chinese Integrative Medicine

基　　金：上海市教育委员会重点学科建设项目(No.J50305);上海市科学技术委员会自然基金资助项目(No.11ZR1436900);上海市教育委员会预算内科研项目(No.2010JW35);上海高校创新团队建设项目(第一期)

摘　　要：目的:研究当药和水飞蓟提取混合物对大鼠在非酒精性脂肪性肝病(non-alcoholicfattyliver disease,NAFLD)基础上由四氯化碳(carbontetrachloride,CCl4)诱导的肝损伤的防治作用及机制。方法:雄性Wistar大鼠随机分为正常组、CCl4组、高脂饮食组、高脂饮食加CCl4组、当药和水飞蓟混合物组(提取物组)、甘草酸二铵组。正常组和CCl4组喂食普通饲料,其余大鼠均给予高脂饲料,提取物组和甘草酸二铵组大鼠同时分别采用当药和水飞蓟混合物或甘草酸二铵干预。8周后,除正常组和高脂饮食组外,其余各组大鼠均腹腔注射小剂量CCl4,注射48 h后收集血清和肝脏组织。苏木精和伊红(hematoxylin and eosin,HE)染色观察肝脏组织病理学改变,检测血清丙氨酸氨基转移酶(alamine aminotrasferas,ALT)、门冬氨酸氨基转移酶(aspartate aminotransferase,AST)活性,肝组织匀浆中三酰甘油(triacylglycerol,TAG)、丙二醛(malondialdehyde,MDA)、谷胱甘肽(glutathione,GSH)含量,并用逆转录聚合酶链反应、蛋白质印迹技术检测解偶联蛋白2(uncoupling protein-2,UCP2)基因和蛋白表达水平。结果:肝组织HE染色显示,小剂量CCl4未导致正常饮食大鼠肝脏出现病理学异常改变,高脂饮食组出现肝小叶大面积肝细胞脂肪变性,高脂饮食加CCl4组脂肪变性加重,肝细胞气球样变和炎性细胞浸润明显,提取物组、甘草酸二铵组肝细胞脂肪变性、气球样变性和炎性浸润明显减轻。与正常组比较,CCl4组大鼠血清ALT、AST活性,肝组织TAG、MDA、GSH水平和UCP2表达无明显改变;各指标除GSH外在高脂饮食组较正常组及CCl4组略有升高;高脂饮食加CCl4组血清ALT、AST水平较高脂饮食组显著升高,肝组织TAG、MDA含量和UCP2 mRNA及蛋白表达显著升高(P<0.05),肝组织GSH含量明显下降;而提取物组和甘草酸二铵组各指标均显著改善(P<0.05)。结论:NAFLD大鼠肝组织对小剂量CCl4损伤敏感性显著增加,肝组�OBJECTIVE： To study the mechanism of liver injury induced by carbon tetrachloride （COl4） in rats with non- alcoholic fatty liver disease （NAFLD）, and the therapeutic effects of the extract mixture of Dangyao （Swertia pseudochinensis Hara） and Shuifeiji （Si/ybum marianum Gaertn） on NAFLD rats with liver injury. METHODS： Male Wistar rats were randomized into normal control group, CCl4 group, highfat diet group, highfat diet plus CCI_4injection group （model group）, diammonium glycyrrhizinate group and extract mixture group. Except the normal control and CCI_4 groups, rats were fed with highfat diet （88% normal chow, 10% lard and 2 % cholesterol） to induce NAFLD. Diammonium glycyrrhizinate and extracts were given by gavage. After eight weeks, a nonlethal dose of CCI4 was injected intraperitoneally to all rats except the normal and high-fat diet groups. And 48 h later, all rats were sacrificed, and serum and liver tissues were collected for further study. Paraffin-processed liver tissue was stained with hematoxylineosin （HE） to observe the pathological changes. Serum aamine aminotrasferas （ALT） and aspartate aminotransferase （AST） were measured. The levels of triacylglycerol （TAG）, malondialdehyde （MDA） and glutathione （GSH） in liver tissues were also examined. Expression of uncoupling protein 2 （UCP2） was determined by reverse transcription polymerase chain reaction and Western blotting. RESULTS： Liver sections stained with HE showed that the histopathological changes in the normal control group and the CCI4 group were mild; massive hepatosteatosis diffusing in Iobules was shown in the high-fat diet groups~ steatosis, hepatocellular ballooning degeneration and inflammatory infiltration were severe around the central vein in sections of the model group. Compared with the model group, hepatosteatosis and ballooning were significantly attenuated in the treatment groups. Levels of serum ALT and AST, contents of TAG and MDA and the UCP2 expression in liver tissues

关 键 词：脂肪肝 非酒精性 肝损伤 四氯化碳 氧化性应激 植物提取物 大鼠 

分 类 号：R259[医药卫生—中西医结合]