目的 基于 PI3K/AKT/BAD 信号通路探讨扶正化纤方对肝纤维化小鼠的作用及机制。方法 采用腹腔注射 20% 四氯化碳-橄榄油溶液 0.2 mL，每周 3 次，连续 4 周，复制肝纤维化小鼠模型。将 C57B/L 雄性小鼠随机分为正常组、模型组、安络化纤丸组 （0.78 g•kg-1） 以及扶正化纤方低、中、高剂量组 （3.1、6.2、12.4 g•kg-1） ，每组 6 只。灌胃给药 （10 mL•kg-1） ，每日 1 次，持续 4 周。使用小动物超声仪检测肝脏弹性值；HE、Masson、天狼星红染色法观察小鼠肝脏组织病理变化；ELISA 法检测血清透明质酸 （HA） 、层黏蛋白 （LN） 、Ⅲ型前胶原（P-ⅢC） 及Ⅳ型胶原 （Ⅳ-C） 水平；qRT-PCR 及 Western Blot 法检测肝脏组织中 AKT、BAD、Bax、Bcl-2 mRNA及蛋白表达；免疫荧光法检测肝脏组织中 α-SMA 表达水平。结果 与空白组比较，模型组小鼠的体质量、肝质量明显降低 （P＜0.05，P＜0.01） ；肝脏弹性值显著升高 （P＜0.01） ；胶原纤维面积占比显著升高 （P＜0.01） ；血清 HA、LN、P-ⅢC、Ⅳ-C 水平均显著升高 （P＜0.01） ；肝脏组织 α-SMA 阳性表达面积占比显著升高 （P＜0.01） ，AKT、Bcl-2 mRNA 及蛋白表达显著上调 （P＜0.01） ，BAD、Bax mRNA 及蛋白表达显著下调 （P＜0.01） 。与模型组比较，各给药组小鼠的体质量、肝质量均明显升高 （P＜0.05，P＜0.01） ；肝脏弹性值均显著降低 （P＜0.01） ；胶原纤维面积占比均显著降低 （P＜0.05，P＜0.01） ；血清 HA、LN、P-ⅢC、IV-C 水平均显著下降（P＜0.01） ；肝脏组织 α-SMA 阳性表达面积占比显著降低 （P＜0.01） ，Bcl-2 mRNA 及蛋白表达显著下调 （P＜0.01） ，AKT 蛋白表达显著下调 （P＜0.01） ，BAD、Bax mRNA 表达显著上调 （P＜0.01） 。扶正化纤方中、高剂量组小鼠肝脏组织中 AKT mRNA 表达显著下调 （P＜0.01） ，Bax 蛋白表达显著上调 （P＜0.01） ；扶正化纤方低、高剂量组小鼠肝脏组织中 BAD 蛋白表达显著上调 （P＜0.01） 。结论 扶正化纤方可能通过 PI3K/AKT/BAD 信号通路调控下游 Bcl-2、Bax 蛋白表达，促进活化肝星状细胞 （HSC） 凋亡，减少细胞外基质 （ECM） 及胶原纤维合成，从而改善四氯化碳诱导的肝纤维化小鼠肝内纤维间质沉积。

Objective To investigate the effect and mechanism of Fuzheng Huaxian Prescription on hepatic fibrosis mice based on PI3K/AKT/BAD signaling pathway. Methods A mouse model of hepatic fibrosis was established by intraperitoneal injection of 0.2 mL CCl4 solution （20% in olive oil） three times per week for four consecutive weeks. C57B/L male mice were randomly divided into normal group，model group，Anluo Huaxian Pills group（0.78 g•kg-1），low-，medium-，and high- dose Fuzheng Huaxian Prescription groups（3.1，6.2，12.4 g•kg-1，respectively），with six mice in each group. The corresponding drug（10 mL•kg-1）was administered by gavage once a day for four weeks. A small-animal ultrasound device was used to measure the value of liver elasticity. Hematoxylin and eosin（HE）staining，Masson’ s trichrome staining and Sirius red staining were used to observe pathological changes in liver tissues. Enzyme-linked immunosorbent assay（ELISA）was used to detect the serum levels of hyaluronic acid （HA），laminin（LN），procollagen III（P-IIIC），and collagen IV （IV-C）. Real-time quantitative polymerase chain reaction （qRT-PCR）and Western Blot were used to measure the mRNA and protein expression levels of protein kinase B （AKT），Bcl-2-associated agonist of cell death （BAD），the anti-apoptotic protein Bcl-2，and the pro-apoptotic protein Bax in liver tissues. Immunofluorescence was used to detect α-SMA expression in liver tissues. Results Compared with the normal group，the body mass and liver mass of rats in the model group were significantly reduced （P＜0.05，P＜0.01）. There was significant increase in the value of liver elasticity （P＜0.01），the proportion of collagen fiber area （P＜0.01），serum levels of HA，LN，P-IIIC，and IV-C（P＜0.01）. The proportion of α-SMA positive expression area in liver tissue significantly increased （P＜0.01）. The mRNA and protein expressions of AKT and Bcl-2 were significantly up-regulated （P＜0.01），while those of BAD and Bax were significantly down-regulated （P＜0.01）. Compared with the model group，the treatment groups exhibited significant increase in the body mass and liver mass（P＜0.05， P＜0.01）， significant reduction in the value of liver elasticity （P＜0.01）， and a decrease in the proportion of collagen fiber area 0.05 0.01 . Serum levels of HA LN P-IIIC and IV-C were significantly reduced （P＜0.01）. The proportion of α -SMA positive expression area in liver tissue significantly decreased（P＜0.01）. Bcl-2 mRNA and protein expression were obviously down-regulated（P＜0.01）. AKT protein expression was significantly down-regulated （P＜0.01）. The mRNA expressions of BAD and Bax were obviously up-regulated （P＜0.01）. AKT mRNA expression in medium- and high- dose Fuzheng Huaxian Prescription groups was significantly down-regulated（P＜0.01），but Bax protein expression was significantly up-regulated（P＜0.01）. BAD protein expression in low- and high- dose Fuzheng Huaxian Prescription groups was remarkably up-regulated （P＜0.01）. Conclusion Fuzheng Huaxian Prescription can significantly improve interstitial deposition of intrahepatic fibrous in hepatic fibrosis mice induced by CCl4，possibly by regulating downstream Bcl-2 and Bax protein expression through PI3K/AKT/BAD signaling pathway，promoting the apoptosis of activated hepatic stellate cells（HSC），as well as reducing extracellular matrix（ECM）and collagen fiber synthesis.

R285.5

沈阳市科学技术计划公共卫生研发专项 （21-173-9-37）；国家中医药管理局项目 （卢秉久全国名老中医药专家传承工作室建设项目，2022-304）