植物雌激素鹰嘴豆芽素A(BCA)对肝纤维化去势小鼠模型的改善作用及机制  

Role and mechanism of action of phytoestrogen biochanin A in improving liver fibrosis in ovariectomized mice

作　　者：谭超容 李小飘 冉俊艳 熊英 廖尚高 张金娟 何迅[1] TAN Chaorong;LI Xiaopiao;RAN Junyan;XIONG Ying;LIAO Shanggao;ZHANG Jinjuan;HE Xun(School of Pharmacy,Guizhou Medical University,Guiyang 550025,China;School of Basic Medicine,Guizhou Medical University,Guiyang 550025,China;Transformation Engineering Research Center of Chronic Disease Diagnosis and Treatment,Guizhou Medical University,Guiyang 550025,China)

机构地区：[1]贵州医科大学药学院,贵阳550025 [2]贵州医科大学基础医学院,贵阳550025 [3]贵州医科大学慢性病诊疗转化工程研究中心,贵阳550025

出　　处：《临床肝胆病杂志》2024年第1期76-82,共7页Journal of Clinical Hepatology

基　　金：贵州省高层次创新型人才“百层次人才”项目(黔科合平台人才[2020]6011)。

摘　　要：目的探究植物雌激素鹰嘴豆芽素A(BCA)对CCl4诱导的雌性双侧卵巢切除(去势)小鼠肝纤维化的影响及其机制。方法取50只去势雌性昆明小鼠,腹腔注射CCl4建立肝纤维化模型,将建模小鼠按体质量随机分成模型组、阳性对照组、BCA低、中、高剂量组各10只,同时选取10只同批雌性小鼠切除双侧卵巢旁少量脂肪组织作为假手术组。假手术组和模型组灌胃等体积的0.5%羧甲基纤维素钠溶液,阳性对照组用雌二醇2 mg/kg灌胃,BCA低、中、高剂量组分别按25、50、100 mg/kg BCA灌胃,1次/d,连续7周。给药结束后麻醉处死小鼠取材,测定肝指数和子宫指数,HE及Masson染色观察肝组织病理改变,生化法检测AST、ALT活性,ELISA法检测肝组织中IL-6、TNF-α水平,Western Blot法测定肝组织中Ⅰ型胶原蛋白(CollagenⅠ)、转化生长因子-β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)、雌激素受体β(ERβ)、p-NF-κBp65/NF-κBp65蛋白的相对表达量。计量资料两组间比较采用成组t检验,多组间比较采用单因素方差分析;进一步两两比较采用LSD-t检验。非正态分布的计量资料多组间比较及进一步两两比较均采用Kruskal-Wallis H检验。结果与假手术组相比,模型组肝指数升高,子宫指数降低,血清AST和ALT活性、肝组织中IL-6、TNF-α水平及CollagenⅠ、TGF-β1、α-SMA、p-NF-κBp65/NF-κBp65蛋白表达均升高(P值均<0.05),肝组织ERβ表达变化不明显(P>0.05),模型组肝组织明显纤维化病变,出现肝细胞水肿、脂肪样变、坏死并伴有炎细胞浸润,胶原纤维增生沉积、交错分布。与模型组比较,BCA各剂量组肝指数降低,血清ALT和AST活性、肝组织IL-6、TNF-α水平及CollagenⅠ、TGF-β1、α-SMA、p-NF-κBp65/NF-κBp65的蛋白表达均降低(P值均<0.05),子宫指数无明显变化(P>0.05),肝组织ERβ蛋白表达升高(P<0.05),肝组织的纤维化病变有不同程度改善。结论BCA可有效改善CCl4诱导的雌性去势小Objective To investigate the effect of phytoestrogen biochanin A(BCA)on liver fibrosis induced by CCl4 in female mice with bilateral oophorectomy(ovariectomized)and its mechanism.Methods A total of 50 ovariectomized Kunming mice were selected and given intraperitoneal injection of CCl4 to establish a model of liver fibrosis,and then according to body weight,they were randomly divided into model group,positive control group,and low-,middle-,and high-dose BCA groups,with 10 mice in each group.In addition,10 female mice in the same litter were given resection of a small amount of adipose tissue near both ovaries to establish the sham-operation group.The mice in the positive control group were given estradiol 2 mg/kg by gavage,and those in the low-,middle-,and high-dose BCA groups were given BCA by gavage at a dose of 25,50,and 100 mg/kg,respectively,once a day for 7 consecutive weeks;the mice in the sham-operation group and the model group were given an equal volume of 0.5%sodium carboxymethyl cellulose solution by gavage.The mice were anesthetized and sacrificed after administration to collect samples.Liver index and uterus index were measured;HE staining and Masson staining were used to observe liver histopathological changes;the biochemical analysis was used to measure the activity of aspartate aminotransferase(AST)and alanine aminotransferase(ALT);ELISA was used to measure the levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in liver tissue,and Western blot was used to measure the relative protein expression levels of collagenⅠ,transforming growth factor-beta 1(TGF-β1),alpha-smooth muscle actin(α-SMA),estrogen receptor beta(ERβ),and p-NF-κBp65/NF-κBp65 in liver tissue.The t-test was used for comparison of continuous data between two groups;a one-way analysis of various was used for comparison between multiple groups,and the least significant difference t-test was used for further comparison between two groups.The Kruskal-Wallis H test was used for comparison of non-normally distributed con

关 键 词：肝纤维化 植物雌激素类 卵巢切除术 小鼠 

分 类 号：R575.2[医药卫生—消化系统] R-332[医药卫生—内科学]