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目的 研究扶正化瘀胶囊(FZHc)对肝纤维化小鼠肝细胞中核因子相关因子2(Nrf2)表达的影响. 方法 将昆明小鼠70只随机分为对照组和模型组.对照组24只,包括正常组(A1)、矿物油组(A2)、FZHc组(A3),每组8只;模型组46只,腹腔注射CCL4,包括模型6周组(B1) 10只、模型10周组(B2) 12只、FZHc低剂量组(C1) 12只、FZHc高剂量组(C2) 12只.其中A3、C1、C2组在第7周起用FZHc药粉(由双蒸水配制成浓度为0.1 g/ml药液)灌胃,连续4周.模型组第6、10周末分批处死小鼠,收集肝脏标本HE染色观察组织炎症程度变化,Masson染色观察纤维化病变;免疫组织化学检测Nrf2、醌氧化还原酶l(Nqol)、α-平滑肌肌动蛋白(α-SMA)、纤维连接蛋白(FN)表达;Western blot检测细胞内Nrf2、Nqol总蛋白表达量及Nrf2核转移情况;RT-PCR检测Nrf2 mRNA表达量.采用SPSS13.0软件进行统计学分析,计量资料采用单因素方差分析,两两比较选用LSD检验,计数资料采用Ridit分析.结果 B1组9只小鼠中有7只(77.8％)为肝纤维化Ⅱ期,有2只小鼠为肝纤维化Ⅲ期.B2组8只小鼠中5只(62.5％)为肝纤维化Ⅲ期,3只为可见假小叶形成.FZHc干预可减轻肝脏炎症及纤维化程度,C1组10只小鼠有6只(60％)为肝纤维化Ⅱ期,4只小鼠肝纤维化Ⅲ期,C2组9只小鼠中有2只(22.2％)为肝纤维化Ⅰ期,5只为肝纤维化Ⅱ期,2只为肝纤维化Ⅲ期,多组间的Kruskal-Wallis检验,x2=53.321,P＜0.01.B2、C1、C2组α-SMA阳性表达平均吸光度值分别为0.09±0.01、0.06±0.01、0.04±0.00;FN阳性表达平均吸光度值分别为0.08±0.01、0.07±0.01、0.05±0.01,F=77.421和F=118.262,P值均＜0.05.不同剂量FZHc干预后α-SMA、FN表达较B2组明显减少,并呈剂量依赖性.B2、C1、C2组小鼠肝组织中Nrf2阳性表达平均吸光度值分别为0.07±0.01、0.10±0.01、0.17±0.01;Nqol平均光吸光度分别为0.06±0.01、0.09±0.01、0.14±0.01,F=182.537和F=75.615,P值均＜0.05;Nrf2总蛋白表达的相对灰度值分别为0.79±0.05、0.94±0.02、1.12±0.08,F=45.664,P＜ 0.05.不同剂量FZHc干预可显著增加Nrf2、Nqol表达,并呈剂量依赖性.C1、C2、B2组Nrf2核蛋白表达的相对灰度值分别为0.97±0.09、1.24±0.10和0.67±0.04,F=94.787,P＜0.05,差异有统计学意义.FZHc促进Nrf2 mRNA相对表达量,C1组为4.00±0.12、C2组为4.65±0.08、B2组为2.15±0.11,F=3230.105,P＜0.05,差异有统计学意义,并具有剂量依赖性. 结论 FZHc可能通过促进Nrf2 mRNA、蛋白表达并促进Nrf2胞核转移,促进其下游靶基因Nqol的表达,抑制HSCs活化及FN合成,从而减轻肝脏纤维化.“,”Objective To investigate the effect of Fuzhenghuayu compound (FZHc) on expression of nuclear factor E2-related factor 2 (Nrf2) in hepatocytes under conditions of hepatic fibrosis using a mouse model.Methods Mice were randomly assigned to a control group and a hepatic fibrosis model group.The control group was further divided into three subgroups for use as normal controls (A1),mineral oil-treated controls (A2),and FZHc-treated controls (A3); the hepatic fibrosis model group was administered carbon tetrachloride (CC14 dissolved in mineral oil and injected intraperitoneally) and further divided into four subgroups for use as 6-weeks models (B1),10-weeks models (B2),low-dose (L)-FZHc models (C1),and high-dose (H)-FZHc models (C2).The FZHc (capsule powder diluted with double-distilled water to 0.1 g/mL) was administered via gastric perfusion to groups A3,C1,and C2 starting at week 7 of the experiment.At the end of week 6 and 10,hepatic specimens were collected and evaluated for degree of hepatic fibrosis and inflammation using routine haematoxylin-eosin staining and Masson staining.Immunohistochemical analysis was performed to measure the hepatocyte expression of Nrf2,NAD(P)H quinine oxidoreductase 1 (Nqol),α-smooth muscle actin (α-SMA) and fibronectin (FN).Real-time fluorescence quantitative PCR was used to measure Nrf2 mRNA expression.western blotting was used to detect Nrf2 and Nqol total protein expression and Nrf2 nuclear translocation.F test,LSD test and ridit test were used for statistical analyses.Results Compared with the B2 group (ridit value:0.09),the model groups treated with FZHc showed significantly lower degrees of hepatic inflammation and fibrosis for both the low (C1 group,ridit value:0.32) and high doses (C2 group,ridit value:0.40) (F =82.927,P ＜ 0.05).In addition,compared with the B2 group,the model groups treated with FZHc showed significantly decreased expression of α-SMA and FN proteins,with a dose-dependent trend (by immunohistochemistry:C 1 group at the end of 10 weeks,F =77.421,118.262,P ＜ 0.05; C2 group,P =0.002,0.013) and significantly increased expression of Nrf2 and Nqol proteins (by immunohistochemistty:C1 and C2 groups at the end of 10 weeks,F =182.537,75.615,P ＜ 0.05 and by westem blotting:F =45.664,127.673,P ＜ 0.05),which also showed a dose-dependent trend (C2 group,P =0.000,0.014; 0.005,0.014).Western blotting also indicated that the amount of nuclear transported Nrf2 was higher in the C1 and C2 groups at the end of 10 weeks (vs.B2 group,F =94.787,P ＜ 0.05),and the amount of nuclear transported Nrf2 was significantly higher in the C2 group (vs.C1 group,P =0.044).Nrf2 mRNA expression was significantly higher in the C1 group than in the B2 group (F =3230.105,P ＜ 0.05),and the C2 group had more substantially increased expression (P =0.001); there was no statistical difference found between groups B1 and B2 (P =0.094).Conclusion Fuzhenghuayu compound increased the expression of Nrf2 mRNA and protein under conditions of hepatic fibrosis in mice and stimulated Nrf2 nuclear transport,as well as increased expression of the Nrf2 target gene Nqol that is known to suppress activation of hepatic stellate cells and decrease the deposition of FN.Therefore,Fuzhenghuayu compound may ameliorate hepatocyte injury in hepatic fibrosis in mice by exerting an antihepatic fibrosis effect.