壮药鸡骨草制剂治疗多房棘球蚴感染诱导的肝纤维化效果  

Treatment effect of Zhuang medicine Herba abri formula on hepatic fibrosis induced by Echinococcus multilocularis infection

作　　者：李嘉静 黄文君[1,2] 曹得萍 LI Jiajing;HUANG Wenjun;CAO Deping(Department of Human Parasitology,Basic Medical College;Key Lab of Pathogenic Biology,Guilin Medical University,Guilin 541199,Guangxi,China)

机构地区：[1]桂林医学院基础医学院人体寄生虫学教研室,广西桂林541199 [2]桂林医学院病原生物学重点实验室,广西桂林541199

出　　处：《中国寄生虫学与寄生虫病杂志》2024年第5期573-581,共9页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：广西自然科学基金(2020GXNSFAA297216)。

摘　　要：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、200μg/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100μg/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 100μg/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�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 200μg/ml HaF solution for 72,96 and 120 h,then the relative expression of α-smooth muscle actin (α-SMA) and collagenⅠin 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 of100μg/ml HaF for 24,48 and 72 h,subsequently,the relative expression of α-smooth muscle actin (α-SMA) and collagenⅠin 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 collagenⅠin 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 con

关 键 词：多房棘球蚴感染 壮药鸡骨草 肝纤维化 

分 类 号：R532.32[医药卫生—内科学]