主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

北京中医药大学学报 ›› 2021, Vol. 44 ›› Issue (1): 45-53.doi: 10.3969/j.issn.1006-2157.2021.01.008

• 中药药理 • 上一篇    下一篇

参苓白术散对脾虚湿困型溃疡性结肠炎大鼠结肠组织TLR2、MyD88、COX-2蛋白和mRNA表达的影响*

李姿慧1,2, 蔡荣林3#, 孙娟1,2, 文印君4, 王瑞4   

  1. 1 安徽中医药大学中医学院 安徽 230012;
    2 新安医学教育部重点实验室;
    3 安徽中医药大学针灸推拿学院;
    4 安徽中医药大学研究生院
  • 收稿日期:2020-04-27 出版日期:2021-01-30 发布日期:2021-01-18
  • 通讯作者: #蔡荣林,男,博士,教授,博士生导师,主要研究方向:中医药防治脾胃病研究,E-mail:ronglincai@163.com
  • 作者简介:李姿慧,女,博士,副教授
  • 基金资助:
    *国家自然科学基金项目(No.81302901),安徽省自然科学基金项目(No.2008085MH298),安徽省高校自然科学研究重点项目(No.KJ2018A0271),安徽中医药大学自然科学研究项目(No.2020zrzd02,No.2016ts008)

Effect of Shenlin Baizhu San on protein and mRNA expression of TLR2, MyD88, COX-2 in ulcerative colitis rats with spleen deficiency and dampness stagnation pattern*

Li Zihui1,2, Cai Ronglin3, Sun Juan1,2, Wen Yinjun4, Wang Rui4   

  1. 1 School of Traditional Chinese Medicine,Anhui University of Chinese Medicine,Anhui 230012,China;
    2 Key Laboratory of Xin An Medicine,Ministry of Education,Anhui University of Chinese Medicine,Anhui 230038,China;
    3 School of Acupuncture and Moxibustion,,Anhui University of Chinese Medicine,Anhui 230012,China;
    4 Graduate School,Anhui University of Chinese Medicine,Anhui 230012,China
  • Received:2020-04-27 Online:2021-01-30 Published:2021-01-18
  • Contact: Prof.Cai Ronglin,Ph.D.,School of Acupuncture and Moxibustion,Anhui University of Chinese Medicine.No.350 Longzihu Road,Xinzhan District,Hefei 230012.E-mail:ronglincai@163.com.
  • Supported by:
    National Natural Science Foundation of China (No. 81302901),Project of Natural Science Foundation of Anhui Province (No. 2008085MH298),Key Projects of Natural Science Research in Universities of Anhui Province (No. KJ2018A0271)

摘要: 目的 观察参苓白术散对脾虚湿困型溃疡性结肠炎(UC)大鼠结肠组织Toll样受体2(TLR2)、髓样分化因子88(MyD88)及环氧化酶-2(COX-2)表达的影响,探讨参苓白术散改善UC的作用机制。方法 采用TNBS/乙醇灌肠结合环境与饮食干预法复制脾虚湿困型UC大鼠模型,按照随机数字表将大鼠分为正常组、模型组、参苓白术散组(15.6 g/kg)、参苓白术散(15.6 g/kg)+ TLR2激动剂(50 μg/只)组、参苓白术散(15.6 g/kg)+ TLR2拮抗剂组(0.121 2μg/g),每组12只,连续给药14 d。ELISA法检测血清肿瘤坏死因子-α(TNF-α)、白细胞介素6(IL-6)及白细胞介素1β(IL-1β)水平,免疫组织化学法和Western Blot法测定结肠组织中TLR2、MyD88、COX-2蛋白表达,Real-time PCR法检测各组大鼠结肠组织TLR2、MyD88、COX-2 mRNA表达情况。结果 模型复制后,脾虚湿困型UC大鼠血清TNF-α、IL-6及IL-1β水平均升高(P<0.05),结肠组织TLR2、MyD88、COX-2蛋白和mRNA表达增强(P<0.01);与模型组比较,参苓白术散组和参苓白术散+TLR2拮抗剂组大鼠血清TNF-α、IL-6及IL-1β水平均下降,结肠组织TLR2、MyD88、COX-2蛋白和mRNA表达下调(P<0.01)。与参苓白术散组比较,参苓白术散+TLR2拮抗剂组大鼠血清TNF-α、IL-6及IL-1β水平,结肠组织TLR2、MyD88、COX-2 蛋白和mRNA表达差异无统计学意义(P>0.05);参苓白术散+TLR2 激动剂组大鼠血清TNF-α、IL-6及IL-1β水平,结肠组织TLR2、MyD88、COX-2 蛋白和mRNA表达均高于参苓白术散组(P<0.01)。结论 参苓白术散可明显下调脾虚湿困型UC大鼠结肠组织TLR2、MyD88、COX-2 蛋白和mRNA表达,降低炎性因子的释放;参苓白术散通过负性调控TLR2/MyD88信号通路,从而减轻肠道炎症反应,是其治疗UC的重要机制之一。

关键词: 参苓白术散, 溃疡性结肠炎, 脾虚湿困, Toll样受体2, 髓样分化因子88, 大鼠

Abstract: Objective To observe the effect of Shenling Baizhu San (Ginseng-Poria-Atractylodes Powder,SLBZS) on the expression of Toll like receptor 2 (TLR2),myeloid differentiation factor 88 (MyD88) and cyclooxygenase-2 (COX-2) in colonic tissue of rats with ulcerative colitis (UC) of spleen deficiency and dampness stagnation pattern,and to explore the mechanism of SLBZS in relieving UC.Methods The model of UC with spleen deficiency and dampness stagnation pattern was reproduced by TNBS/ethanol enema combined with environment at and dietarg intervention.Rats were randomly divided into five groups:normal group,model group,SLBZS group,SLBZS+ TLR2 agonist (Pam3csk4) group,and SLBZS+ TLR2 antagonist (T2.5) group (n=12 in each group for continuous intervention of 14 days).The levels of TNF-α,IL-6 and IL-1β in the serum were detected by ELISA,the expression of TLR2,MyD88 and COX-2 protein in colon tissue was measured with immunohistochemical method and Western Blot Assay,and the expression of TLR2 mRNA,MyD88 mRNA and COX-2 mRNA in colon tissue of rats in each group was measured by using real time PCR.Results The levels of TNF-α,IL-6 and IL-1β in the serum of UC rats with spleen deficiency and dampness stagnation pattern were increased (P<0.05),and the expressions of TLR2,MyD88,COX-2 and TLR2 mRNA,MyD88 mRNA and COX-2 mRNA in the colon tissue were increased (P<0.01).After the treatment of SLBZS,the levels of TNF-α,IL-6 and IL-1β in the serum of UC rats were decreased,and the expression of TLR2,MyD88,COX-2 protein and mRNA were decreased,compared with the model group (P<0.01).Compared with SLBZS group,there was no difference in the levels of TNF-α,IL-6 and IL-1β in the serum,and the expression of TLR2,MyD88,COX-2 protein and mRNA in colon tissue,in SLBZS+TLR2 antagonist group (P>0.05).Compared with SLBZS group,the levels of TNF-α,IL-6 and IL-1β in the serum,and the expression of TLR2,MyD88,COX-2 protein and mRNA in colon tissue of SLBZS+TLR2 agonist group were higher (P<0.01).Conclusion SLBZS may significantly decrease the expression of TLR2,MyD88,COX-2 protein and mRNA in colon tissue of UC rats with spleen deficiency and dampness stagnation pattern,and reduce the release of inflammatory factors.It is one of the important mechanisms of SLBZS in the treatment of UC by negatively regulating TLR2/MyD88 signal pathway to reduce intestinal inflammation.

Key words: Shenling Baizhu Powder, ulcerative colitis, spleen deficiency and dampness stagnation pattern, toll-like receptor 2, myeloid differentiation factor 88, rats

中图分类号: 

  • R285.5