扶正化瘀方及其治法拆方对肝损伤小鼠肝细胞凋亡的干预作用  

Effects of Chinese herbal medicine Fuzheng Huayu recipe and its components against hepatocyte apoptosis in mice with hepatic injury

作　　者：周滔[1,2,3] 闫秀川[1,2] 陈倩[1,2] 陶艳艳[1,2] 胡义扬[1,2] 刘平[1,2] 刘成海[1,2,4,5] 

机构地区：[1]上海中医药大学曙光医院 [2]上海中医药大学肝病研究所,上海201203 [3]首都医科大学附属北京中医医院消化中心,北京100010 [4]上海高校中医内科学E-研究院,上海201203 [5]上海市中医临床重点实验室,上海201203

出　　处：《中西医结合学报》2011年第1期57-63,共7页Journal of Chinese Integrative Medicine

基　　金：国家重点基础研究发展计划(973计划)资助项目(No.2006CB504801);国家自然科学基金资助项目( No.30772869);上海教育委员会创新团队资助项目;上海市优秀学科带头人计划资助项目(No.08XD14041);上海市教育委员会E-研究院建设项目(No. E-03008)

摘　　要：目的:观察扶正化瘀方治法拆方对小鼠肝细胞凋亡的影响,并探讨扶正化瘀方影响肝细胞凋亡的治法特点与作用机制。方法:BABL/c雄性小鼠50只,随机分为正常组、模型组、扶正化瘀方组、扶正拆方组和化瘀拆方组,每组各10只。采用脂多糖(10μg/kg)联合半乳糖胺(900mg/kg)腹腔注射诱导急性肝损伤。各用药组小鼠于造模前3d开始分别给予扶正化瘀方、扶正拆方和化瘀拆方灌胃,每日2次,连续3d,第4日灌胃后1h开始造模;正常组与模型组小鼠给予等量生理盐水灌胃。采用比色法检测血清丙氨酸氨基转移酶(alanine aminotransferase,ALT)、天冬氨酸氨基转移酶(aspartate aminotransferase,AST)、超氧化物歧化酶(super-oxide dimutase,SOD)活性和丙二醛(malondialdehyde,MDA)含量;苏木精-伊红染色观察肝组织炎症病理改变;原位末端标记法观察肝组织肝细胞凋亡情况;荧光定量实时聚合酶链式反应法检测肝组织肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)mRNA表达;蛋白印迹法检测肿瘤坏死因子Ⅰ型受体(tumor necrosis factor receptor typeⅠ,TNFR1)蛋白表达。结果:与模型组比较,扶正化瘀方及其拆方药物扶正拆方、化瘀拆方均可显著降低模型小鼠血清ALT和AST活性,改善肝组织炎症,减轻肝细胞凋亡,改善肝组织过氧化指标SOD活性和MDA含量,其中全方作用最好,扶正拆方优于化瘀拆方。与正常组比较,模型组肝组织TNF-αmRNA表达显著升高,扶正化瘀方及扶正拆方、化瘀拆方可显著降低其表达,其中全方作用最好,两拆方组相比差异无统计学意义。与正常组比较,模型组肝组织TNFR1蛋白表达显著升高,全方可显著降低其表达,而两拆方无明显作用。结论:扶正化瘀方和其治法拆方中扶正方可抑制小鼠肝细胞凋亡,扶正化瘀方抗肝细胞凋亡作用机制可能在于减轻肝脏脂质过氧化损伤和下调TNF-α及TNFR1的表达。Objective：To investigate the effects and characteristics of Fuzheng Huayu recipe,a compound Chinese herbal medicine,and its decomposed therapies against hepatocyte apoptosis in mice with hepatic injury.Methods：A total of 50 male BALB/c mice were randomly divided into 5 groups：control group,untreated group,Fuzheng Huayu recipe group,Fuzheng recipe group and Huayu recipe group. Hepatocyte apoptosis in mice was induced by intraperitoneal injection of lipopolysaccharide （10 μg/kg） and galactosamine （900 mg/kg）. The mice in drug-treated groups were administered with Fuzheng Huayu recipe,Fuzheng recipe and Huayu recipe by garvage respectively 3 days before injection of lipopolysaccharide and galactosamine. The mice were sacrificed 6 hours after the administration of lipopolysaccharide and galactosamine. The activities of alanine aminotransferase （ALT）,aspartate aminotransferase （AST） and superoxide dimutase （SOD） and content of malondialdehyde （MDA） were examined by colorimetric method. Inflammation and necrosis in liver tissue were observed with hematoxylin-eosin staining. Hepatocyte apoptosis was determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining. Protein expression of tumor necrosis factor receptor type Ⅰ （TNFR1） was analyzed with Western blotting. Expression of tumor necrosis factor-α （TNF-α） mRNA was analyzed with real-time fluorescent quantitative polymerase chain reaction.Results：Compared with the untreated group,Fuzheng Huayu recipe,Fuzheng recipe and Huayu recipe attenuated hepatocyte apoptosis,decreased the serum ALT and AST activities and MDA content,and improved the SOD activity in liver tissues. Of the three groups,the effect of Fuzheng Huayu recipe group was the best,Fuzheng recipe group was better than Huayu recipe group. Compared with the untreated group,all drugs exerted good effects in decreasing TNF-α mRNA expression,and Fuzheng Huyu recipe was better than the others. The expression of TNFR1 protein in the untreat

关 键 词：肝细胞 凋亡 中草药 拆方研究 脂质过氧化作用 受体 肿瘤坏死因子 Ⅰ型 小鼠 

分 类 号：R285.5[医药卫生—中药学]