吉林大学学报(医学版) ›› 2022, Vol. 48 ›› Issue (2): 391-398.doi: 10.13481/j.1671-587X.20220216

• 基础研究 • 上一篇    下一篇

毛蕊异黄酮对肝硬化大鼠肠黏膜屏障功能的影响及其机制

刘琦(),徐新,王正根   

  1. 南华大学附属第二医院消化内科,湖南 衡阳 421001
  • 收稿日期:2021-07-12 出版日期:2022-03-28 发布日期:2022-05-10
  • 通讯作者: 刘琦 E-mail:sbxz6329@163.com
  • 作者简介:刘 琦(1983-),男,湖南省衡阳市人,主治医师,医学硕士,主要从事消化道疾病基础和临床方面的研究。主治医师(E-mail:sbxz6329@163.com
  • 基金资助:
    湖南省卫健委科研项目(20201932)

Effect of calycosin on intestinal mucosal barrier function in cirrhosis rats and its mechanism

Qi LIU(),Xin XU,Zhenggen WANG   

  1. Department of Gastroenterology,Second Affiliated Hospital,South China University,Hengyang 421001,China
  • Received:2021-07-12 Online:2022-03-28 Published:2022-05-10
  • Contact: Qi LIU E-mail:sbxz6329@163.com

摘要: 目的

探讨毛蕊异黄酮对肝硬化大鼠肠黏膜炎症和屏障功能的影响,并阐明其作用机制。

方法

采用四氯化碳(CCl4)联合乙醇复合法诱导大鼠肝硬化模型,将造模成功的36只大鼠随机分为模型组、低剂量毛蕊异黄酮组(5 mg·kg-1)和高剂量毛蕊异黄酮组(20 mg·kg-1),每组12只,另选12只SD大鼠作为对照组。低和高剂量毛蕊异黄酮组大鼠给予相应药物灌胃,对照组和模型组大鼠给予等量生理盐水灌胃,每天1次,连续4周。采用生物化学法检测各组大鼠血清中丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶(AST)活性,HE染色观察各组大鼠肝脏和回肠组织病理形态表现,ELISA法检测各组大鼠血清中D-乳酸(D-LA)、二胺氧化酶(DAO)和内毒素(ET)水平及回肠组织中白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)水平,Western blotting法检测各组大鼠回肠组织中Toll样受体4(TLR4)、核因子κB p65(NF-κB p65)和NF-κB抑制因子α(IκBα)蛋白表达水平。

结果

与对照组比较,模型组大鼠肝脏和回肠组织损伤严重,血清中ALT和AST活性,D-LA、DAO和ET水平均明显升高(P<0.05),回肠组织中IL-1β、TNF-α和IL-6水平,TLR4和NF-κB p65蛋白表达水平均明显升高(P<0.05),IκBα蛋白表达水平明显降低(P<0.05);与模型组比较,高剂量毛蕊异黄酮组大鼠肝脏和回肠组织损伤明显改善,血清中ALT和AST活性,D-LA、DAO和ET水平均明显降低(P<0.05),回肠组织中IL-1β、TNF-α和IL-6水平及TLR4和NF-κB p65蛋白表达水平均明显降低(P<0.05),IκBα蛋白表达水平明显升高(P<0.05);与模型组比较,低剂量毛蕊异黄酮组大鼠以上各指标差异无统计学意义(P>0.05)。与低剂量毛蕊异黄酮组比较,高剂量毛蕊异黄酮组大鼠肝脏和回肠组织损伤明显改善,血清中ALT和AST活性及D-LA、DAO和ET水平均明显降低(P<0.05),回肠组织中IL-1β、TNF-α和IL-6水平及TLR4和NF-κB p65蛋白表达水平均明显降低(P<0.05),IκBα蛋白表达水平明显升高(P<0.05)。

结论

毛蕊异黄酮能够改善肝硬化大鼠肠黏膜屏障损伤,其机制可能与抑制TLR4/NF-κB信号通路有关。

关键词: 毛蕊异黄酮, 肝硬化, 肠黏膜屏障, TLR4/NF-κB信号通路

Abstract: Objective

To investigate the effect of calycosin on the intestinal mucosal inflammation and barrier function in the cirrhosis rats, and to elucidate its mechanism.

Methods

The rat models of cirrhosis were induced by carbon tetrachoride (CCl4) combined with ethanol complex method.A total of 36 cirrhosis rats were randomly divided into model group, low dose of calycosin group (5 mg·kg-1) and high dose of calycosin group (20 mg·kg-1), with 12 rats in each group,and another 12 SD rats were used as control group.The rats in low and high dose calycosin groups were given corresponding drugs by gavage,and the rats in control group and model group were given equal amount of saline by gavage,once a day, for consecutive 4 weeks. The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum of the rats in various groups were determined by biochemistry method, HE staining was used to observe the pathomorphology of liver and ileum tissues of the rats in various groups.The levels of D-lactic acid (D-LA), diamine oxidase (DAO) and endotoxin (ET) in serum and the levels of interleukin-1β (IL-1β), tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6) in ileum tissue were detected by ELISA. The expression levels of Toll-like receptor 4 (TLR4), nuclear factor kappa B p65 (NF-κB p65) and NF-κB inhibitor α (IκBα) proteins in ileum tissue of the rats in various groups were detected by Western blotting method.

Results

Compared with control group, severe damage to the liver and ileum tissue in model group was revealed, the activities ALT, AST and the levels of D-LA, DAO and ET in serum were significantly increased (P<0.05), the levels of IL-1β, TNF-α and IL-6 and the expression levels of TLR4 and NF-κB p65 proteins in ileum tissue were significantly increased (P<0.05), while the expression level of IκBα protein was significantly decreased (P<0.05). Compared with model group, the damage of liver and ileum tissue of the rats in high dose of calycosin group was significantly improved, the activities of ALT, AST and the levels of D-LA, DAO and ET in serum were significantly decreased (P<0.05), the levels of IL-1β, TNF-α, IL-6 and the expression levels of TLR4 and NF-κB p65 proteins in ileum tissue were significantly decreased (P<0.05), and the expression level of IκBα protein was significantly increased (P<0.05);compared with model group, there were no statistical differences in the above indexes in low dose of calycosin group(P>0.05). Compared with low dose of calycosin group, the damage of liver and ileum tissue of the rats in high dose of calycosin group was significantly improved, the activities of ALT, AST and the levels of D-LA, DAO and ET in serum were significantly decreased (P<0.05), the levels of IL-1β, TNF-α, IL-6 and the expression levels of TLR4 and NF-κB p65 proteins in ileum tissue were significantly decreased (P<0.05), and the expression level of IκBα protein was significantly increased (P<0.05).

Conclusion

Calycosin can improve the damage of intestinal mucosal barrier in the cirrhosis rats, and its mechanism may be related to inhibition of TLR4/NF-κB signaling pathway.

Key words: Calycosin, Cirrhosis, Intestinal mucosal barrier, Toll-like receptor 4/Nuclear factor kappa B signaling pathway

中图分类号: 

  • R574.53