一贯煎治疗二甲基亚硝胺致小鼠肝纤维化的机制研究

Efficacy studies of Yiguanjian treatment in mice fibrosis induced by dimethylnitrosamine

关键词:  一贯煎  HGF  肝纤维化  DMN肝纤维化小鼠  SDF-1  CXCR4  Wnt-1  β-Catenin  NF-κB p65  ERK1+2

Key words:  [Key words]　Yiguanjian  HGF  liver fibrosis  DMN hepatic fibrosis in mice  SDF-1  CXCR4  Wnt-1  β-Catenin  NF-κB p65  ERK1+2

摘要:

[摘要]　目的　探讨一贯煎改善小鼠肝纤维化的作用机制。　方法　将205只小鼠分为模型组185只和正常对照组20只，模型组腹腔注射二甲基亚硝胺（DMN）4周制备肝纤维化模型，正常对照组腹腔注射同等量生理盐水。于造模成功后将模型组随机分为3组：一贯煎组、阳性对照组（肝细胞生长因子组即HGF组）、空白组，一贯煎组予一贯煎灌胃给药，HGF组予肝细胞生长因子皮下注射给药，空白组予生理盐水皮下注射治疗，共治疗4周。分别于造模2周、4周、治疗2周、4周时观察小鼠一般状况、血清生化肝功、肝组织HE染色及免疫组化染色并进行半定量分析，评估一贯煎对肝纤维化的治疗作用及对相关因子的影响。　结果　（1）一贯煎治疗后小鼠腹水等症状逐渐改善，死亡率降低；（2）模型组4周时ALT、AST、TBIL、DBIL、ALP均较正常对照组明显升高，ALB、CHE较正常对照组明显降低；一贯煎组ALT和AST均在第2周时明显降低； ALB在第2周明显增高（P=0.001）； CHE在第4周明显增高（P=0.000），且第4周较第2周明显增加（P=0.001）； ALP在第4周时明显降低（P=0.037）；TBIL、DBIL、GGT在治疗前后差异无明显变化。(3) 肉眼观察一贯煎组治疗后部分肝脏轻度肿大，黄褐色，表面尚光滑，HE染色肝组织结构较完整，肝细胞变性、坏死情况较模型组轻。一贯煎组4周时α-SMA表达弱阳性，较空白组强阳性表达减弱。(4)SDF-1/CXCR4及ERK1+2均为一贯煎组4周阳性表达较2周明显增高； Wnt-1/β-catenin及NF-κB p65均为一贯煎组4周阳性表达较2周明显降低。　结论　一贯煎能改善小鼠肝纤维化和肝功能，SDF-1/CXCR4、Wnt-1/β-catenin、NF-κB p65、ERK1+2参与小鼠肝纤维化过程，一贯煎通过调控上述机制发挥作用。

Abstract:

[Abstract]　Objective　To investigate effective way of Yiguanjian to improve the process of liver fibrosis in mice. 　Methods　205 mice were divided into 185 rats named model group and 20 rats named normal control group, liver fibrosis model was prepared with dimethylnitrosamine, the model group were randomly divided into 3 groups after modeling: Yiguanjian group, positive control group (hepatocyte growth factor group, HGF group)and blank control group. The group treated with Yiguanjian Decoction were treated with intragastric administration.HGF group was treated with hepatocyte growth factor by subcutaneous injection, the control group were treated with normal saline by subcutaneous injection, the total treatment was four weeks. The general condition of mice, serum biochemistry, HE staining， tissue immunohistochemistry were appllied to detect effect of YGJ, differentiation and related factors affected in modeling for 2 weeks, 4 weeks and treatment for 2 weeks,4 weeks. 　Results　（1）With Yiguanjian treatment, the a small amount of ascites symptoms and so on were improved, the mortality rate decreased gradually.(2) Model group 4 weeks of liver function in the ALT, AST, TBIL, DBIL, ALP were markedly elevated compared with normal control group (P<0.05), A1B, CHE was lower than the control group (P<0.05).After Yiguanjian treatment， ALT and AST decreased significantly in the fourth week than that in the second week;ALB increased significantly during the second week（P=0.001）; CHE in the fourth week did increase significantly higher（P=0.000）than that in the model， and the fourth week there was a significant increase compared with the second week (P=0.001); ALP did decrease significantly in the fourth week（P=0.037）;TBIL, DBIL, GGT before and after the treatment had no significant meaning. (3) It could be detected in YGJ group like slightly enlarged part of the liver， brown and smooth surface,which were more complete microscopic structure of the liver tissue and less liver cell degeneration and necrosis compared with the model group.α -SMA expression of Yiguanjian group 4 weeks was weak positive, compared with blank treatment group with strongly positive expression. (4)The positive staining of SDF-1/CXCR4 and ERK1+2 in Yiguanjian group 4 week were more than that in Yiguanjian group 2 weeks. The positive staining of Wnt-1/β-catenin and NF- κB p65 in Yiguanjian group 2 week was less than that in Yiguanjian group 4 week. 　Conclusion　YGJ could improve the efficacy of liver fibrosis. A variety of signal factor such as SDF-1/CXCR4, Wnt-1/β-catenin, NF-κB p65, ERK1+2 were refered in liver fibrosis reversal process. Yiguanjian play a role through the above mechanisms.