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 ›› 2020, Vol. 43 ›› Issue (5): 402-407.doi: 10.3969/j.issn.1006-2157.2020.05.009

• 中药药理 • 上一篇    下一篇

逍遥散对非酒精性脂肪性肝炎大鼠Toll样受体4基因甲基化水平的调节作用*

徐慧超, 高艳, 陈浩, 郝健亨, 刘晋芳, 刘杨, 苗宇船#   

  1. 山西中医药大学基础医学院 山西 030619
  • 收稿日期:2019-11-17 发布日期:2020-06-08
  • 通讯作者: #苗宇船,女,博士,教授,硕士生导师,主要研究方向:肝病的病因病机理论研究,E-mail:mych65@163.com
  • 作者简介:徐慧超,女,在读硕士生
  • 基金资助:
    *国家自然科学基金资助项目(No.81470190),山西省卫生健康委员会科研项目(No.2019089),山西省研究生教育创新项目(No.2019SY510),山西省“1331工程”立德树人“好老师”课程建设计划项目(No.18),山西省研究生教育改革研究项目(No.2019JG190),山西中医药大学“肝脏炎性疾病中西医结合基础研究创新团队”项目(No.2018TD-016)

Treatment of nonalcoholic steatohepatitis by regulating the methylation level of TLR4 gene with Xiaoyao San*

Xu Huichao, Gao Yan, Chen Hao, Hao Jianheng, Liu Jinfang, Liu Yang, Miao Yuchuan#   

  1. School of Basic Medicine, Shanxi University of Chinese Medicine, Shanxi 030619, China
  • Received:2019-11-17 Published:2020-06-08
  • Contact: Prof. Miao Yuchuan, Ph.D., Master’s Supervisor. School of Basic Medicine, Shanxi University of Chinese Medicine, Jinzhong 030619, China. E-mail: mych65@163.com
  • Supported by:
    National Natural Science Foundation of China (No.81470190); Scientific Research Project of Health Commission of Shanxi Province (No.2019089)Ethical review: Committee of Medical and Animal Ethical Review on Experimental Animals in Shanxi University of Chinese Medicine (No.2019LL016)

摘要: 目的 探讨逍遥散调节Toll样受体4(TLR4)基因甲基化水平治疗非酒精性脂肪性肝炎(non-alcoholic steatohepatitis,NASH)肝郁脾虚证大鼠的机制。方法 将30只雄性SD大鼠随机分为正常组、模型组及逍遥散组。正常组大鼠给予正常饮食28周,后4周注射生理盐水;模型组与逍遥散组大鼠饲喂高脂高糖饮食28周,并通过慢性应激束缚及饥饱失常刺激制备肝郁脾虚证模型,后4周分别注射生理盐水及逍遥散溶液。28周末收集3组大鼠尿液,取肝脏及海马组织,制作病理切片、采用实时荧光定量PCR(qPCR)、酶联免疫吸附实验(ELISA)、亚硫酸氢测序PCR(BSP)等方法检测相应指标。结果 与模型组相比,逍遥散组大鼠肝郁脾虚证症状明显缓解,证候积分降低;肝细胞内的脂肪空泡及炎性细胞浸润程度显著减少;肝组织TLR4 mRNA含量下降,TLR4基因甲基化水平升高(P<0.05)。结论 逍遥散可能通过上调非酒精性脂肪性肝炎肝郁脾虚证大鼠模型中TLR4基因甲基化水平,下调其TLR4 mRNA的表达,从而改善肝脏脂肪变性程度、炎症反应及肝郁脾虚症状,这可能是逍遥散治疗非酒精性脂肪性肝炎肝郁脾虚证的机制。

关键词: 逍遥散, 非酒精性脂肪性肝炎, 肝郁脾虚证, DNA甲基化, Toll样受体4

Abstract: Objective To explore the new mechanism of changes in TLR4 gene methylation levels in the treatment of liver stagnation and spleen deficiency pattern in non-alcoholic steatohepatitis (NASH) in rats with Xiaoyao San (Xiaoyao Powder). Methods 30 male SD rats were randomly divided into blank group, model group and Xiaoyao San group (n=10). Rats in the blank group were given normal diet for 28 weeks. followed by injection of normal saline for 4 weeks. In the model group and Xiaoyao San group, liver stagnation and spleen deficiency pattern model was established by feeding the rats with a high-fat and high-sugar diet with stimulations of chronic stress and irregular diets for 28 weeks In the last 4 weeks, the model group and Xiaoyao San group were injected with normal saline and Xiaoyao San solution respectively. At the end of 28 weeks, urine was collected from the three groups, and the liver and hippocampus tissues were taken to make pathological sections. The corresponding indicators were detected by real-time fluorescence quantitative PCR (qPCR), enzyme-linked immunosorbent assay (ELISA), and hydrogen sulfite sequencing PCR (BSP), etc. Results Compared with the model group, the symptoms of liver stagnation and spleen deficiency pattern were significantly alleviated with a decreased pattern score in the Xiaoyao San group. The numbers of fat vacuoles and infiltration degree of inflammatory cells in liver cells were significantly reduced. The TLR4 mRNA content in liver tissues was decreased, while the methylation level of TLR4 gene was increased (P<0.05). Conclusion Xiaoyao San may up-regulate the TLR4 gene methylation level and down-regulate the TLR4 mRNA expression in rats with liver stagnation and spleen deficiency pattern in non-alcoholic steatohepatitis, thus improving the degree of liver steatosis and alleviating inflammation and other symptoms. This seems to be the new mechanism for the treatment of such condition with Xiaoyao San.

Key words: Xiaoyao San (Xiaoyao Powder), nonalcoholic steatohepatitis (NASH), liver stagnation and spleen deficiency pattern, DNA methylation, TLR4

中图分类号: 

  • R285.5