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 ›› 2018, Vol. 41 ›› Issue (9): 759-770.doi: 10.3969/j.issn.1006-2157.2018.09.010

• 科技之窗 • 上一篇    下一篇

基于转录组测序技术研究黄芪水煎液对虚寒证大鼠免疫机能的影响*

于婉晨, 王世军#, 季旭明, 赵海军, 宗鑫, 陈艳娇   

  1. 山东中医药大学 山东 250355
  • 收稿日期:2018-04-10 出版日期:2018-09-30 发布日期:2018-09-30
  • 通讯作者: 王世军,男,博士,教授,博士生导师,主要研究方向:脑及肿瘤微循环的研究及中药药性研究,E-mail:pathology@163.com
  • 作者简介:于婉晨,女,在读博士生
  • 基金资助:
    国家重点基础研究发展规划(973计划) 项目(No. 2013CB531803), 国家自然科学基金项目(No. 81673852、81703838、81603419), 山东省自然科学基金项目(No. ZR2016HM66), 山东省重点产业关键技术项目(No. 2016CYJS08A01-3、2016CYJS08A01-4), 山东省重点研发计划项目(No. 2016ZDJS07A12)

Influence of water decoction of Huangqi on immune function in rats with deficiency cold pattern: a study based on transcriptome sequencing technique*

Yu Wanchen, Wang Shijun#, Ji Xuming, Zhao Haijun, Zong Xin, Chen Yanjiao   

  1. Shandong University of Traditional Chinese Medicine, Shandong 250355, China
  • Received:2018-04-10 Online:2018-09-30 Published:2018-09-30
  • Supported by:
    National Key Basic Research Program of China (973 Program, No. 2013CB531803), National Natural Science Foundation of China (No. 81673852, No. 81703838, No. 81603419), Natural Science Foundation of Shandong Province (No. ZR2016HM66), Key Technology Project of Major Industry in Shandong Province (No. 2016CYJS08A01-3, 2016CYJS08A01-4), Key R&D Plan of Shandong Province (No. 2016ZDJS07A12)

摘要: 目的 探讨虚寒证的发生及黄芪水煎液对虚寒证产生治疗作用的分子机制。方法 选用传统中药复方(生石膏、龙胆草、黄柏、知母)按照2∶1.2∶1∶1.5的比例,灌胃给药16 g/(kg·d),持续14 d,建立虚寒证大鼠模型。模型建立后,采用SPSS软件随机分为虚寒模型组及黄芪水煎液组,黄芪水煎液组给予5.40 g/(kg·d)灌胃给药,连续7 d,应用转录组测序技术检测空白组、虚寒模型组、黄芪水煎液组大鼠肝脏基因表达,筛选差异表达基因,根据GO功能标准,对免疫相关基因进行注释,荧光定量PCR对测序结果进行验证。结果 与空白组相比,虚寒模型组筛选出差异基因323条;与虚寒模型组相比,黄芪水煎液组筛选出差异基因333条,主要涉及刺激应答与防御应答功能。GO功能分析显示,虚寒模型大鼠出现以刺激、防御应答为代表的多种免疫应答相关基因表达的下调,影响机体的免疫应答功能,削弱机体免疫;给予黄芪水煎液治疗后,通过上调刺激、防御应答相关基因表达,进一步改善机体对外界的免疫应答情况,使机体免疫状态得以增强、恢复。结论 虚寒证发生及黄芪水煎液对虚寒证治疗作用的分子机制可能与机体免疫机能及其相关基因表达密切相关。

关键词: 黄芪水煎液, 虚寒证, 免疫机能, 转录组测序, 大鼠

Abstract: Objective To investigate the molecular mechanism of occurrence of deficiency cold pattern and therapeutic effect of water decoction of Huangqi (Milkvetch Root, Radix Astragali seu Hedysari) on deficiency cold pattern. Methods The traditional Chinese medical compound [Sheng Shigao (raw Gypsum, Gypsum Fibrosum), Longdancao (Gentian Root, Radix Gentianae), Huangbai (Amur Cork-Tree, Cortex Phellodendri), Zhimu (Common Anemarrhena Rhizome, Rhizoma Anemarrhenae), in a ratio of 2∶1.2∶1∶1.5] was intragastrically given to rats [dosage=16 g/(kg·d), and dose=4 mL/(kg·d)] for 14 d for establishing rat model of deficiency cold pattern. The modeled rats were randomly divided into model group(deficiency model) and treatment group [treated with water decoction of Huangqi, dose=5.40 g/(kg·d), each n=12] by using SPSS software, and other rats (n=12) were chosen into blank group. The expressions of immune-related genes in liver were detected by using transcriptome sequencing technique. The differentially expressed genes (DEG) were screened, and immune-related genes were annotated according to GO functional standard. The Results of transcriptome sequencing were verified by using fluorescence quantitative polymerase chain reaction (PCR). Results Compared with blank group, there were 323 differential genes screened in model group. Compared with model group, there were 333 differential genes screened in treatment group, and they mainly involved the functions of stimulus response and defense response. The Results of GO functional analysis showed that model group had down-regulated expressions of multiple immune-related genes with stimulus response and defense response as representations, and immune response function was impacted and body immunity was impaired. After treatment with water decoction of Huangqi, the immune response of body to environment was improved and body immune status was recovered and enhanced through up-regulating the expressions of genes related to stimulus response, defense response and immune response. Conclusion The molecular mechanism of occurrence of deficiency cold pattern and therapeutic effect of water decoction of Huangqi may closely related to body immune function and expressions of relative genes.

Key words: water decoction of Huangqi (Milkvetch Root, Radix Astragali seu Hedysari), deficiency cold pattern, immune function, transcriptome sequencing, rats

中图分类号: 

  • R285.5