杭白菊提取液对小鼠肝纤维化的保护作用及机制研究

Protective effect and mechanism of Dendranthema morifolium on hepatic fibrosis in mice

目的探讨杭白菊对肝纤维化的保护作用及作用机制。方法 SPF级小鼠72只随机分成6组。灌胃用药:正常对照组和模型对照组为0.9%NaCl注射液,水飞蓟宾组为5.5mg/mL水飞蓟宾,杭白菊低、中、高浓度组分别为杭白菊提取液150、300、600mg/mL;腹腔注射用药:除正常对照组为0.9%NaCl注射液外,其余均为0.1%四氯化碳(CCl_4)摘除眼球法采血检测小鼠血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、透明质酸(HA)、白细胞介素-6(IL-6)、超氧化物歧化酶(SOD)、丙二醛(MDA),取肝检测肝指数,取肝右叶行病理检测。结果模型对照组的血清ALT、AST、HA、IL-6、MDA均明显高于正常对照组(P<0.01),而肝指数明显低于正常对照组(P<0.01),肝细胞结构模糊,肝索排列紊乱甚至消失,肝细胞变性坏死区域明显由结缔组织代之。水飞蓟宾组的血清ALT、AST、HA、IL-6、SOD、MDA与正常对照组比较差异无统计学意义(P>0.05),而明显低于模型对照组(P<0.01,P<0.05)。杭白菊低、中、高浓度组的血清ALT、AST、HA(除杭白菊高浓度组)和MDA明显高于正常对照组而明显低于模型对照组(P<0.01);杭白菊低、中、高浓度组血清IL-6与正常对照组和模型对照组比较差异均无统计学意义(P>0.05);杭白菊低浓度组的血清SOD明显高于模型对照组(P<0.01),杭白菊高浓度组的血清SOD明显高于模型对照组和正常对照组(P<0.01,P<0.05);杭白菊中、高浓度组的血清HA明显低于杭白菊低浓度组(P<0.01,P<0.05),杭白菊高浓度组的MDA明显低于杭白菊低、中浓度组(P<0.01,P<0.05)。模型对照组、水飞蓟宾组和杭白菊低、中、高浓度组的肝指数之间的两两比较差异均无统计学意义(P>0.05)但均明显低于正常对照组(P<0.05,P<0.01)。水飞蓟宾组和杭白菊低、中浓度组的肝细胞结构较模型对照组清晰,肝细胞变性坏死区域有较大的改善;正常对照组和杭白菊高浓度组的肝细胞形态正常,肝索排列规则,肝细胞无明显变性坏死。结论 CCl_4生成自由基和炎性反应造成肝纤维化;水飞蓟宾和杭白菊通过消除自由基和炎性反应干预肝纤维化;高浓度杭白菊干预肝纤维化效果优于水飞蓟宾,由此推测杭白菊干预肝纤维化可能还存在其他途径。

Objective To investigate the protective effect and mechanism of Dendranthema morifolium on hepatic fibrosis in mice.Methods Seventy-two SPF mice were randomly divided into 6 groups:the normal control group(0.9%NaCl injection),model control group(0.9%NaCl injection),silybin group(5.5 mg/mL silybin),low,medium and high concentration of Dendranthema moritolium(150,300,600 mg/mL Dendranthema moritolium extract)group,each group was intragastrically administered with the corresponding drugs or solvents;except for the normal control group which intraperitoneal injection of 0.9%NaCl,the other were intraperitoneal injection of 0.1%carbon tetrachloride(CCl 4);Both intragastrically administion and intraperitoneal injection amount was 10 mL/kg.The mice in each group were intragastrically administed once a day for a week,then intragastrically administion and intraperitoneal injection for 4 weeks alternately,and then intragastrically administed again once a day for a week.The ALT,AST,HA,IL-6,SOD and MDA were detected through blood sampling at 30th min after the last gavage.The liver index was measured and the pathological examination was performed on the right lobe of the liver.Results The serum ALT,AST,HA,IL-6 and MDA in the model control group were significantly higher than those in the normal control group(P<0.01),while the liver index was significantly lower than that in the normal control group(P<0.01),the structure of hepatocytes was blurred,the arrangement of hepatic cord was disordered or even disappeared,and the degeneration and necrosis area of hepatocytes was obviously replaced by connective tissue.There was no difference of the serum levels of ALT,AST,HA(except for high concentration of Dendranthema group),IL-6,MDA in silybin group than in the normal control group(P>0.05),but it was lower than that in the model control group(P<0.01,P<0.05),there was no significant difference in HA between high concentration group and normal control group(P>0.05).There was no statistical difference in serum IL-6 between low,middle and high concentration groups of Dendranthema morifolium group and normal control group and model control group(P>0.05).The serum level of SOD in the low concentration group was higher than that in the model control group(P<0.01).The serum SOD in the high concentration of Dendranthema morifolium group was significantly higher than that in the model control group and the normal control group(P<0.01,P<0.05).The serum HA in the medium and high concentration group of Dendranthema morifolium group was significantly lower than that in the low concentration group(P<0.01,P<0.05),and the MDA in the high concentration of Dendranthema morifolium group was significantly lower than that of low and middle concentration groups(P<0.01,P<0.05).There was no significant difference in liver index between model control group,silybin group and low,medium,high concentration of Dendranthema morifolium group(P>0.05),but it was significantly lower than that of normal control group(P<0.05,P<0.01).The hepatocyte structure in the silybin group and middle concentration group was more clear than that in the model control group,and the area of hepatocyte necrosis was improved greatly.In the normal control group and the high concentration group Dendranthema morifolium,the morphology of hepatocytes was normal,the arrangement of hepatic cord was regular,and there was no obvious degeneration and necrosis of hepatocytes.Conclusion CCl 4 produces free radicals and inflammation to cause liver fibrosis;silybin and Dendranthema morifolium interfere with liver fibrosis by eliminating free radicals and inflammatory responses,there may be other ways of preventing hepatic fibrosis by Dendranthema morifolium.