壮药牛大力醇提物对四氯化碳诱导肝纤维化大鼠的改善作用研究  

Improvement effects of ethanol extract from Zhuang medicine Millettia speciosa Champ.on carbon tetrachloride-induced hepatic fibrosis in rats

作　　者：侯玲 覃妮 陆世银 陈秋贵 磨奕玲 徐敏丽 卢鲜云 何烨 张洪平 HOU Ling;QIN Ni;LU Shiyin;CHEN Qiugui;MO Yiling;XU Minli;LU Xianyun;HE Ye;ZHANG Hongping(Graduate School,Guangxi University of Chinese Medicine,Nanning,Guangxi 530000,China;Department of Pharmacy,Liuzhou Municiple Liutie Central Hospital,Liuzhou,Guangxi 545000,China;Key Laboratory of Research and Development of Chinese Medicine(Zhuang&Yao Medicine)Preparations,Liuzhou Traditional Chinese Medicine Hospital(Liuzhou Hospital of Zhuang Medicine),Liuzhou,Guangxi 545026,China)

机构地区：[1]广西中医药大学研究生院,广西南宁530000 [2]柳州市柳铁中心医院,药剂科,广西柳州545000 [3]柳州市中医医院(柳州市壮医医院),中药(壮瑶药)制剂研发重点实验室,广西柳州545026

出　　处：《湖南中医药大学学报》2025年第1期15-22,共8页Journal of Hunan University of Chinese Medicine

基　　金：广西中医药大学自然科学基金面上项目(2020MS055);柳州市科技攻关与新产品试制(2020NBAA0802)。

摘　　要：目的研究牛大力醇提物对四氯化碳(CCl4)诱导的肝纤维化大鼠的治疗作用及机制。方法采用腹腔注射40%CCl4橄榄油溶液(0.1 mL/100 g)建立肝纤维化SD大鼠模型,2次/周,连续12周。将肝纤维化大鼠随机分为模型组,牛大力醇提物低、中、高剂量(3.5、7、14 g/kg)组及阳性对照秋水仙碱(0.12 mg/kg)组,另设正常组大鼠,各组均为8只。各给药组于造模第9周按设定剂量灌胃相应药物干预,1次/d,连续4周。正常组与模型组大鼠每天灌胃等体积的生理盐水。末次给药24 h后,测定大鼠肝脾指数;检测各组血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、丙二醛(MDA)、超氧化物歧化酶(SOD)、透明质酸(HA)、层粘连蛋白(LN)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原(Ⅳ-C)、转化生长因子β1(TGF-β1)、白细胞介素-1β(IL-1β)、白细胞介素-8(IL-8)及白细胞介素-6(IL-6)的水平;HE染色和Masson染色观察肝组织病理学变化;Western blot测定大鼠肝脏中Kelch样环氧氯丙烷相关蛋白-1(Keapl)、核转录因子红系2相关因子2(nuclear factor-erythroid 2-related factor 2,Nrf2)、血红素加氧酶-1(HO-1)和醌氧化还原酶1(NQO1)的蛋白表达水平。结果与正常组相比,模型组大鼠反应迟钝,肝脾指数、血清ALT、AST、MDA、LN、HA、PCⅢ、Ⅳ-C、TGF-β1、IL-1β、IL-8及IL-6水平显著升高(P<0.01),SOD表达水平显著下降(P<0.01);肝组织呈现明显纤维化;肝组织Keap1蛋白含量明显升高(P<0.01),Nrf2、HO-1和NQO1蛋白表达水平显著降低(P<0.01)。与模型组比较,牛大力各剂量组均能明显改善大鼠状态,肝脾指数及血清ALT、AST、MDA、LN、HA、PCⅢ、Ⅳ-C、TGF-β1、IL-1β、IL-8及IL-6水平显著下调(P<0.05,P<0.01),SOD的表达水平明显增加(P<0.01);有效改善大鼠肝组织病理学损伤和纤维增生;肝组织中Keap1表达被显著抑制(P<0.01),Nrf2、HO-1和NQO1蛋白含量明显升高(P<0.05,P<0.01)。与秋水仙碱组相比,牛大力醇提物高剂量组对�Objective To investigate the therapeutic effects and mechanism of Millettia speciosa Champ.ethanol extract(MCEE)on carbon tetrachloride(CCl4)-induced hepatic fibrosis(HF)in rats.Methods An SD rat model with HF was established by intraperitoneal injection of 40%CCl4 olive oil solution(0.1 mL/100 g)twice a week for 12 consecutive weeks.HF rats were randomly divided into model group,low-,medium-,and high-dose MCEE(3.5,7,14 g/kg)groups,positive control colchicine(0.12 mg/kg)group,and normal group,with eight rats in each group.Each medication group was given the corresponding drug intervention by gavage at the set dose since the 9th week of modeling,once a day for four consecutive weeks.The normal group and model group were given an equal volume of normal saline by gavage every day.The liver and spleen indexes of rats were measured 24 hours after the last administration;serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),malondialdehyde(MDA),superoxide dismutase(SOD),hyaluronic acid(HA),laminin(LN),typeⅢprocollagen(PCⅢ),collagenⅣ(Ⅳ-C),transforming growth factor-β1(TGF-β1),interleukin-1β(IL-1β),interleukin-8(IL-8),and interleukin-6(IL-6)were determined;HE staining and Masson staining were used to observe the pathological changes in liver tissue;Western blot was used to measure the protein levels of Kelch-like ECH-associated protein-1(Keap1),nuclear factor-erythroid 2-related factor 2(Nrf2),heme oxygenase-1(HO-1),and quinone oxidoreductase 1(NQO1)in rat liver.Results Compared with the normal group,rats in the model group showed delayed response,significantly increased liver and spleen indexes,and serum levels of ALT,AST,MDA,LN,HA,PCⅢ,Ⅳ-C,TGF-β1,IL-1β,IL-8,and IL-6(P<0.01),as well as significantly decreased SOD levels(P<0.01);they also exhibited obvious fibrosis in liver tissue,significantly increased Keap1 protein content in liver tissue(P<0.01),and significantly decreased protein levels of Nrf2,HO-1,and NQO1(P<0.01).Compared with the model group,the condition of rats in MCEE

关 键 词：牛大力 醇提物 肝纤维化 抗氧化 炎症因子 Keap1/Nrf2信号通路 

分 类 号：R285.5[医药卫生—中药学]