安络化纤丸对肝纤维化大鼠肝组织基质金属蛋白酶及其抑制物表达的影响  

Effect of Anluohuaxianwan on the expression of matrix metalloproteinases and their inhibitors in rat liver with fibrosis

作　　者：王林 卢玮 高玉华 曹曦 裴斐[2] 刘学恩 庄辉 Wang Lin;Lu Wei;Gao Yuhua;Cao Xi;Pei Fei;Liu Xueen;Zhuang Hui(Department of Microbiology and Center of Infectious Diseases,School of Basic Medical Sciences,Peking University Health Science Center,Beijing 100191,China;Department of Pathology,School of Basic Medical Sciences,Peking University Health Science Center,Beijing 100191,China)

机构地区：[1]北京大学医学部基础医学院病原生物学系和感染病中心,100191 [2]北京大学医学部基础医学院病理学系,100191

出　　处：《中华肝脏病杂志》2019年第4期267-273,共7页Chinese Journal of Hepatology

基　　金：北京大学医学部与森隆药业有限公司的合作项目.

摘　　要：目的用四氯化碳(CCl4)诱导大鼠肝纤维化模型,探索安络化纤丸对肝组织中基质金属蛋白酶(MMPs)及其组织抑制物(TIMPs)表达的影响。方法36只Wistar雄性大鼠随机分为对照组、模型组和治疗组,模型组和治疗组大鼠腹腔注射40%CCl4(2ml/kg),对照组大鼠腹腔注射等渗盐水,每周2次,共6周。建模的同时,治疗组大鼠灌胃安络化纤丸溶液(浓度为0.15g/ml,9.9ml/kg),其他两组大鼠灌胃等渗盐水,每天1次,共6周。第3和第6周末检测血清中丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)。第6周末时采集肝组织,用于病理组织学评价和MMP-2/13、TIMP-1/2的mRNA和蛋白表达水平检测。据资料不同分别用采用单因素方差分析、LSD法、非参数检验中的Kruskal-WallisH检验、Mann-WhitneyU检验进行统计学分析。结果与模型组相比,安络化纤丸显著减轻了治疗组大鼠肝损伤,表现为大鼠一般状态、肝脏和脾脏形态、肝脏和脾脏指数、ALT和AST水平的改善。肝组织病理学诊断表明,治疗组大鼠肝纤维化程度较模型组显著改善(2.75±0.75比3.55±0.69,P=0.015)。治疗组大鼠肝组织中MMP-13的mRNA和蛋白相对表达水平显著高于模型组(mRNA:10.50±7.64比4.40±2.97,P=0.029;蛋白:1.15±0.09比0.78±0.21,P=0.016),而MMP-2和TIMP-1/2的mRNA和蛋白相对表达水平显著低于模型组(mRNA:4.55±3.29比7.83±4.19,P=0.048;1.66±0.73比3.69±2.78,P=0.023;2.25±1.16比3.41±1.51,P=0.049;蛋白:0.44±0.11比0.65±0.05,P=0.03;0.69±0.06比1.07±0.21,P=0.016;0.46±0.09比0.81±0.13,P=0.003)。结论安络化纤丸发挥抗肝纤维化作用主要通过改善肝功能、抑制肝星状细胞的激活、增强MMP-13的表达、抑制MMP-2和TIMP-1/2的表达等实现。Objective To investigate the effect of anluohuaxianwan (ALHXW) using rat model of carbon tetrachloride (CCl4) induced liver fibrosis on the expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs).Methods Thirty-six male Wistar rats were randomly assigned into control,model and treatment groups.Rats in the model and treatment groups were injected intraperitoneally with 40% CCl4 (2 ml/kg),and the control group were given isotonic saline twice a week for six weeks.Meanwhile,the treatment group were gavaged with ALHXW solution daily (concentration 0.15 g/ml,9.9 ml/kg) for 6 weeks,while the control and model groups were given isotonic saline once a day for 6 weeks.Serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured at the end of third and sixth week.At the end of six weeks,liver tissues were harvested for histopathological evaluation and the detection of mRNA and protein expression levels of MMP-2/13 and TIMP-1/2.According to different data,LSD method,parametric (one-way ANOVA) and non-parametric tests (Kruskal-Wallis H-test and Mann-Whitney U test) were used for statistical analysis.Results Compared with the model group,ALHXW markedly alleviated liver injury in the treatment group,and thereby improved the general state of rats,liver and spleen morphological characteristics,and ALT and AST levels.Histopathological examination demonstrated that the extent of liver fibrosis was improved (2.75 ± 0.75 vs.3.55 ± 0.69,P = 0.015) in the treatment group as compared with the model group.The mRNA and protein expression levels of MMP-13 in the treatment group were significantly higher than that of the model group (mRNA: 10.50 ± 7.64 vs.4.40 ± 2.97,P = 0.029.Protein: 1.15 ± 0.09 vs.0.78 ± 0.21,P = 0.016),whereas the mRNA and protein expression levels of MMP-2,TIMP-1/2 in the treatment group were significantly lower than that of the model group (mRNA: 4.55 ± 3.29 vs.7.83 ± 4.19,P = 0.048;1.66 ± 0.73 vs.3.69 ± 2.78,P = 0.023;2.25 ± 1

关 键 词：肝纤维化 四氯化碳 安络化纤丸 基质金属蛋白酶 金属蛋白酶组织抑制物 

分 类 号：R285.5[医药卫生—中药学]