目的 观察体内和体外壮药鸡骨草制剂（HaF）治疗多房棘球蚴感染诱导的肝纤维化效果。 方法 取鸡骨草胶囊制备25、50、100、200 µg/ml HaF溶液，分别体外处理人肝星状细胞LX2（HSC-LX2）72、96、120 h后，蛋白质免疫印迹（Western blotting）检测在HSC-LX2中α-平滑肌肌动蛋白（α-SMA）和Ⅰ型胶原蛋白（collagen Ⅰ）的相对表达量。取0.1 mg/ml多房棘球蚴虫体蛋白刺激HSC-LX2 24 h，加入100 µg/ml HaF溶液处理24、48、72 h后，Western blotting检测HSC-LX2中α-SMA和collagen Ⅰ相对表达量。将36只雌性昆明小鼠随机分为对照组（4只）、感染组（4只）、0.5 g/（kg•d）HaF治疗组（5只）、1.0 g/（kg•d）HaF治疗组（5只）、2.0 g/（kg•d）HaF治疗组（6只）、阿苯达唑（ABZ）治疗组（6只）、ABZ + HaF联合治疗组（6只）等7个组，感染组和各治疗组小鼠分别腹腔注射2 000个原头节，对照组注射等体积生理盐水。感染后60 d，治疗组小鼠灌胃相应药物，1次/d。治疗60 d后，取小鼠肝、脾组织称重，计算小鼠肝脏指数和脾脏指数；碱水解法测定肝组织羟脯氨酸（HYP）含量；比色法测定血清天冬氨酸转氨酶（AST）水平；Western blotting检测肝组织α-SMA和collagen Ⅰ相对表达量；苏木精-伊红（HE）染色、Masson染色观察小鼠肝、脾组织病理变化。两组间数据比较采用独立样本t检验。 结果 100 µg/ml HaF溶液干预96 h后，HSC-LX2中α-SMA和collagen Ⅰ相对表达量分别为0.401 ± 0.218、0.352 ± 0.058，均低于对照组（1.435 ± 0.297、1.340 ± 0.416）（t = 2.755、11.120，均P < 0.05）；其他浓度和作用时间与对照组差异无统计学意义（均P > 0.05）。经多房棘球蚴虫体蛋白刺激的HSC-LX2在HaF处理48 h后的α-SMA和collagen Ⅰ相对表达量分别为0.326 ± 0.106和0.315 ± 0.076，均低于对照组（0.895 ± 0.417、1.009 ± 0.378）（t = 6.359、9.059，均P < 0.05）。体内实验结果显示，感染组小鼠的肝脏指数、脾脏指数、血清AST水平和肝组织HYP含量分别为（4.366 ± 0.284）%、（5.129 ± 1.114）mg/g、（22.194 ± 1.509）U/L和（21.743 ± 2.503）× 10^-2 μg/mg，均高于对照组的（3.389 ± 0.045）%、（2.031 ± 0.165）mg/g、（17.355 ± 0.574）U/L和（9.330 ± 1.519）× 10^-2 μg/mg（t = 3.393、2.752、2.616、4.549，均P < 0.05）。0.5 g/（kg•d）治疗组、1.0 g/（kg•d）治疗组、2.0 g/（kg•d）HaF治疗组、ABZ治疗组、ABZ + HaF联合治疗组小鼠血清AST水平分别为（17.375 ± 0.746）、（15.411 ± 1.338）、（17.057 ± 1.066）、（16.190 ± 1.559）、（14.637 ± 1.888）U/L，均低于感染组（t = 2.863、3.362、2.838、2.717、3.002，均P < 0.05），其中1.0 g/（kg•d）HaF治疗组降低最明显；5个治疗组小鼠肝脏指数、脾脏指数、肝组织HYP含量与感染组比较差异均无统计学意义（均P > 0.05）。Western blotting结果显示，1.0 g/（kg•d）HaF治疗组小鼠肝组织中α-SMA相对表达量为0.818 ± 0.195，ABZ + HaF联合治疗组小鼠肝组织collagen Ⅰ相对表达量为0.799 ± 0.167，均低于感染组（1.668 ± 0.131、1.341 ± 0.141）（t = 3.619、2.486，均P < 0.05）。HE染色和Masson染色表明，ABZ + HaF联合治疗组小鼠肝纤维化程度明显改善，肝组织中纤维组织面积为（6.184 ± 1.951）%，低于感染组的（19.916 ± 2.046）%（t = 4.703，P < 0.01）。 结论 壮药鸡骨草制剂在体内和体外均可明显减轻多房棘球蚴感染诱导的肝纤维化，为鸡骨草胶囊作为多房棘球蚴病的新型治疗制剂提供了实验依据。

Objective To observe the treatment effect of Zhuang medicine Herba abri formula (HaF) on hepatic fibrosis induced by Echinococcus multilocularis infection in vivo and in vitro. Methods The HaF solution was prepared from Zhuang medicine Herba abri capsule commercial available. Human hepatic stellate cell-LX2 (HSC-LX2) were cultured with 25, 50, 100 or 200 µg/ml HaF solution for 72, 96 and 120 h, then the relative expression of α-smooth muscle actin (α-SMA) and collagen Ⅰ in HSC-LX2 was detected with Western blotting, respectively. Separately, HSC-LX2 cells were stimulated with 0.1 mg/ml E. multilocularis crude antigen for 24 h, followed by addition of 100 μg/ml HaF for 24, 48 and 72 h, subsequently, the relative expression of α-smooth muscle actin (α-SMA) and collagen Ⅰ in HSC-LX2 were detected by Western blotting. Thirty-six female Kunming mice were randomly assigned into the following groups: control group (4 mice), infection group (4 mice), HaF treatment groups at doses of 0.5 g/(kg•d) (5 mice), 1.0 g/(kg•d) (5 mice), and 2.0 g/(kg•d) (6 mice), albendazole (ABZ) treatment group (6 mice), and ABZ + HaF combined treatment group (6 mice). The infection group and each treatment group received intraperitoneal injection of 2 000 protoscolices, while the control group was injected with an equal volume of normal saline. After a period of 60 days post-infection, the mice in treatment groups were treated daily by gavage with the corresponding medication for 60 days, and then the mice liver and spleen tissues were collected and weighed to calculate hepatic index and splenic indix. The content of hydroxyproline (HYP) in liver tissue was determined by the alkaline hydrolysis method, and the level of serum aspartate aminotransferase (AST) was measured by the colorimetric method. Western blotting was performed to detect the relative expression of α-SMA and collagen Ⅰ in the liver, while hematoxylin-eosin staining (HE) and Masson staining were used to observe pathological changes in liver and spleen tissues. The comparison between two groups of data was conducted using an independent-samples t-test. Results The relative expression of α-SMA protein and collagen Ⅰ was 0.401 ± 0.218 and 0.352 ± 0.058, respectively, in HSC-LX2 cells treated with 100 µg/ml HaF solution at 96 h, which were significantly lower than those in the control group (1.435 ± 0.297, 1.340 ± 0.416) (t = 2.755, 11.120; both P < 0.05). The control group did not exhibit any statistically significant differences compared to the other concentrations and durations of treatment. Compared to the control group (0.895 ± 0.417, 1.009 ± 0.378), the relative expression levels of α-SMA and collagen Ⅰ in HSC-LX2 treated with HaF for 48 h after E. multilocularis crude antigen stimulation significantly decreased to (0.326 ± 0.106) and (0.315 ± 0.076), respectively (t = 6.359, 9.059; both P < 0.05). In vivo experiments showed that the hepatic indices, splenic indices, serum AST levels and liver HYP contents of infection group were (4.366 ± 0.284) %, (5.129 ± 1.114) mg/g, (22.194 ± 1.509) U/L, and (21.743 ± 2.503) × 10^-2 μg/mg respectively, which were higher than (3.389 ± 0.045) %, (2.031 ± 0.165) mg/g, (17.355 ± 0.574) U/L and (9.330 ± 1.519) × 10^-2 μg/mg in the control group (t = 3.393, 2.752, 2.616, 4.549; all P < 0.05). The serum AST levels of mice in the HaF treatment group at doses of 0.5, 1.0, and 2.0 g/(kg·d), the ABZ treatment group, and the combined ABZ + HaF treatment group were (17.375 ± 0.746), (15.411 ± 1.338), (17.057 ± 1.066), (16.190 ± 1.559), (14.637 ± 1.888) U/L, respectively, lower than those in infection group (t = 2.863, 3.362, 2.838, 2.717, 3.002; all P < 0.05). Among the treatment groups, the HaF treatment group at a dosage of 1.0 g/(kg•d) showed the most significant reduction in serum AST levels. There were no significant differences in hepatic/splenic indices and HYP content in liver tissue between the five treatment groups and the infection group (all P > 0.05). Western blotting results showed that the relative expression level of α-SMA protein in the liver tissue of mice in the 1.0 g/(kg•d) HaF treatment group was 0.818 ± 0.195, and the relative expression level of collagen Ⅰ in the liver tissue of mice in the combined treatment group of ABZ and HaF was 0.799 ± 0.167, both lower than those in the infected group (1.668 ± 0.131, 1.341 ± 0.141) (t = 3.619, 2.486; P < 0.05). The results of HE staining and Masson staining demonstrated that the fibrotic tissue area in the ABZ + HaF combined treatment group was (6.184 ± 1.951)%, which was significantly reduced compared to (19.916 ± 2.046)% in the infection group (t = 4.703, P < 0.01), indicating a notable improvement in liver fibrosis. Conclusion Zhuang medicine HaF demonstrates the ability to attenuate hepatic fibrosis induced by E. multilocularis infection both in vitro and in vivo. It also provides an experimental basis for HaF as a novel therapeutic agent for treatment of alveolar echinococcosis.