七味白术散对HRV dsRNA诱导乳鼠机体表达IFN-1a/β的调节作用及抗病毒效应

Qiweibaizhu Powder on HRV the dsRNA induced suckling mice expression of. IFN-1 alpha / beta regulation and antiviral effects of.（Antiviral Effect and Impact of Qiwei Baizhu Powder on TRL3/RIG1 Signal Transduction Pathway Components of HRV Infected Suckling Mice）..Rotavirus infectious diarrhea is a serious threat to infants and young children. Chinese herbal prescription Qiwei Baizhu Powder (QWBZP) has protective effects on lymphocytes and intestine mucosal organizations between the HRV infected suckling mice's small intestine mucosa. However, there has been no report about how this prescription identifies and eliminates HRV by regulating the innate immune system both at home and abroad. This research experiments upon HRV infected suckling mice applying comparison method and molecular biology technology to observe the effect of QWBZP on TRL3/RIG1 recognition of HRV and the regulatory role of QWBZP in the downstream components of TRL3/RIG1 signal pathway. Besides, research explores the effects of QWBZP on the expression of inflammatory factor, nuclear transcription factor genes as well as IRF3, IRF7, ISRE, IPS1 and MAVS proteins. In addition, it analyzes the effect of QWBZP on the expression of IFN-1(α/β) which is the final effector molecule in HRV infected suckling mice's mononuclear cells and detects the mechanism of QWBZP on HRV elimination. All of these will reveal the inherent antiviral mechanism of QWBZP-TRL3/RIG1-downstream components-IFN-1(α/β) and find the antiviral targets of QWBZP, which will provide further theoretical evidence for treating infantile viral diarrhea with traditional Chinese medicine.

轮状病毒（HRV）感染腹泻是严重威胁婴幼儿生命的疾病。中药七味白术散对HRV感染乳鼠小肠黏膜间淋巴细胞及肠粘膜组织有很好的保护作用。但该方如何通过调节固有免疫系统来识别并清除HRV？目前国内外尚无报道。为此采用C57/BL与TRL3-/-乳鼠感染HRV，用比对法和细胞分子生物学技术：观察该方对感染机体中TRL3/RIG1识别HRV的特征，对TRL3/RIG1信号通路下游元件的调控影响；探讨药物在TRL3/RIG1信号通路中对细胞炎性因子、核转录因子基因与IRF3、IRF7、ISRE、IPS1、MAVS蛋白的表达影响；分析药物对HRV感染乳鼠单个核细胞内RL3/RIG1终末效应分子IFN-1(α/β)表达，及清除HRV的作用机制。揭示七味白术散- TRL3/RIG1-下游信号元件- IFN-1(α/β)抗病毒的内在机理，与药物抗病毒作用靶点，为中医药治疗婴幼儿病毒性腹泻提供依据有着重要意义

研究背景.内容HRV感染性腹泻是婴幼儿重要死因之一，目前临床无特殊治疗药物。在国家基金委和湖南省基金委的资助下，对中药七味白术散治疗HRV的机理在实验乳鼠和实验细胞中进行了探讨，发现该方能抗HRV，更重要的是该方对感染机体有很好的免疫调节作用。HRV是一种dsRNA病毒侵入机体后为什么会引起婴幼儿腹泻？是否与机体的天然有免疫有关（即TRL3对dsHRV识别和下游分子的反应有关）？为此再次研究：中药七味白术散对HRV dsRNA诱导乳鼠机体表达IFN1α/β的调节作用及抗病毒效应。.结果 1.发现HRV导致腹泻的主要原因是病毒dsRNA内吞体进入细胞能逃逸宿主免疫TRL3的识别，使机体大量细胞炎性因子的产生，引起病理损伤。采用激动剂、抑制剂和七味白术散进行比对研究：发现七味白术散对细胞Toll样受体有较好的调节作用，能降低TRL3，下调细胞信号通路中炎性因子表达，促进抑炎因子及其终末效应分子IFN-1(α/β)、IFN-γ表达，减轻感染机体的炎症反应，上调机体抗病毒蛋白表达从而清除HRV。这一研究为精方治疗病毒性腹泻提供了新的理论依据。2..iEL是一类分布在不同的上皮组织内效应性和记忆性淋巴细胞。分布于肠黏膜上皮细胞间iEL通过TCR特异性识别MHC-抗原肽，活化、增殖，分泌抗病毒效应分子抗感染。研究发现： HRV感染乳鼠小肠黏膜上皮内淋巴细胞内淋巴细胞IRF3、IRF7表达受阻，使下游分子IFN及其抗病毒蛋白不能启动转录，七味白术散治疗后能增加IRF3、IRF7表达，从而启动干扰素基因的转录，促使抗病毒蛋白合成，达到抗HRV保护肠粘膜的作用。3.七味白术散有促进肠道正常菌群的恢复作用 提取七味白术散中各药物有效组分进行小鼠血浆药代动力学分析经过高效液相质谱分析，研制出能够抗病毒，迅速恢复肠道正常菌群的七味白术散口服滴剂，初步解决了婴幼儿服药困难问题.（正在申报专利）。