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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

谭超容 李小飘 冉俊艳 熊英 廖尚高 张金娟 何迅

引用本文:
Citation:

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

DOI: 10.12449/JCH240114
基金项目: 

贵州省高层次创新型人才“百层次人才”项目 (Guizhou Kehe Platform Talents [2020] 6011)

伦理学声明:本课题实验方案于2021年7月9日获得贵州医科大学实验动物伦理委员会审批,批号:2100313,符合实验室动物管理与使用准则。
利益冲突声明:本文不存在任何利益冲突。
作者贡献声明:谭超容参与课题的设计,负责课题实施,数据分析,撰写论文;李小飘、冉俊艳、熊英、廖尚高参与课题实施,数据收集,修改论文;张金娟、何迅负责课题设计,指导实验实施及文章撰写并定稿。
详细信息
    通信作者:

    张金娟, 826609585@qq.com (ORCID: 0000-00002-9078-531X)

    何迅, 2812878586@qq.com (ORCID: 0000-0002-1867-069X)

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

Research funding: 

Guizhou Provincial High-Level Innovative Talents “Hundred Level Talents” Project (Guizhou Kehe Platform Talents [2020] 6011)

More Information
  • 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(Collagen Ⅰ)、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Collagen Ⅰ、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Collagen Ⅰ、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诱导的雌性去势小鼠肝纤维化,其作用机制可能是通过上调ERβ抑制NF-κB信号通路,减轻炎症反应而实现的。

     

  • 图  1  BCA对CCl4诱导肝纤维化小鼠子宫、肝脏形态学及肝脏病理的影响

    Figure  1.  Effects of BCA on the morphological and pathological changes of uterus and liver in CCl4-induced liver fibrosis mice

    图  2  BCA对CCl4诱导肝纤维化小鼠蛋白表达的影响

    Figure  2.  Effects of BCA on protein expressions in CCl4-induced liver fibrosis mice

    表  1  BCA对CCl4诱导肝纤维化小鼠肝、子宫指数的影响

    Table  1.   Effects of BCA on liver and uterus weight ratios in CCl4-induced liver fibrosis mice

    组别 动物数(只) 肝指数 子宫指数
    假手术组 6 3.63%±0.35% 0.34%(0.27%~0.58%)
    模型组 6 5.29%±0.47%1) 0.09%(0.05%~0.34%)1)
    阳性对照组 6 4.97%±0.25% 0.49%(0.45%~0.65%)2)
    BCA低剂量组 6 4.51%±0.43%2) 0.11%(0.06%~0.33%)
    BCA中剂量组 6 4.54%±0.44%2) 0.08%(0.05%~0.32%)
    BCA高剂量组 6 4.64%±0.20%2) 0.10%(0.09%~0.11%)
    统计值 F=19.942 H=24.059
    P <0.000 1 <0.000 1
    注:与假手术组比较,1)P<0.05;与模型组比较,2)P<0.05。
    下载: 导出CSV

    表  2  BCA对CCl4诱导肝纤维化小鼠AST、ALT活性的影响

    Table  2.   Effects of BCA on serum activities of AST and ALT in CCl4-induced liver fibrosis mice

    组别 动物数(只) AST(U/L) ALT(U/L)
    假手术组 10 14.92±2.21 7.81±0.61
    模型组 10 43.65±17.711) 32.44±9.141)
    阳性对照组 10 20.56±5.322) 14.96±6.232)
    BCA低剂量组 10 21.17±3.872) 17.22±4.032)
    BCA中剂量组 10 20.72±6.482) 16.70±5.882)
    BCA高剂量组 10 19.66±4.682) 14.97±2.962)
    F 13.703 19.488
    P <0.000 1 <0.000 1
    注:与假手术组比较,1)P<0.05;与模型组比较,2)P<0.05。
    下载: 导出CSV

    表  3  BCA对CCl4诱导肝纤维化小鼠IL-6、TNF-α水平的影响

    Table  3.   Effects of BCA on serum contents of IL-6 and TNF-α in CCl4-induced liver fibrosis mice

    组别 动物数(只) IL-6(pg/mg) TNF-α(pg/mg)
    假手术组 10 107.43±21.23 37.01±12.16
    模型组 10 368.39±76.411) 94.49±26.511)
    阳性对照组 10 123.40±43.222) 43.05±11.702)
    BCA低剂量组 10 208.26±37.812) 70.77±9.782)
    BCA中剂量组 10 172.18±55.652) 53.43±14.382)
    BCA高剂量组 10 113.15±17.512) 40.63±17.442)
    F 31.623 12.645
    P <0.000 1 <0.000 1
    注:与假手术组比较,1)P<0.05;与模型组比较,2)P<0.05。
    下载: 导出CSV

    表  4  BCA对CCl4诱导肝纤维化小鼠肝组织中蛋白表达的影响

    Table  4.   Effects of BCA on protein expressions in CCl4-induced liver fibrosis mice

    组别 动物数(只) α-SMA TGF-β1 Collagen Ⅰ ERβ p-NF-κBp65/NF-κBp65
    假手术组 3 0.15±0.02 0.28±0.06 0.31±0.14 0.63±0.12 0.45±0.13
    模型组 3 0.39±0.051) 0.72±0.071) 0.60±0.021) 0.59±0.13 0.90±0.211)
    阳性对照组 3 0.26±0.102) 0.45±0.062) 0.42±0.032) 0.94±0.272) 0.41±0.182)
    BCA低剂量组 3 0.26±0.082) 0.56±0.092) 0.47±0.022) 0.83±0.15 0.64±0.12
    BCA中剂量组 3 0.19±0.062) 0.49±0.082) 0.43±0.022) 0.95±0.152) 0.45±0.092)
    BCA高剂量组 3 0.18±0.032) 0.47±0.062) 0.42±0.032) 0.98±0.152) 0.53±0.082)
    F 5.850 12.057 6.895 3.207 5.018
    P <0.006 <0.0001 <0.003 <0.046 <0.01
    注:与假手术组比较,1)P<0.05;与模型组比较,2)P<0.05。
    下载: 导出CSV
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  • 收稿日期:  2023-03-09
  • 录用日期:  2023-03-29
  • 出版日期:  2024-01-23
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