桦褐孔菌多糖对弓形虫感染宿主细胞TLRs/MyD88信号通路干预机制研究

Toxoplasmosis is a kind of worldwide pandemic amphixenosis caused by Toxoplasm gondii which parasitize in nucleated cells of various kinds of animals and people. It seriously impairs mankind health and animal husbandry development. We have analyzed biologic pathway of differential gene by KEGG database and DAVID bioinformatics analysis after polysaccharide from Inonotus obliquus treated mice, and dicovered that polysaccharide from Inonotus obliquus could regulate Toll receptor signal pathway. Based on thise preliminary work, the in vitro effects of polysaccharide from Inonotus obliquus on key protein expressions and phosphorylation levels, nuclear transcription factor AP-1 and NF-κB activities, MyD88 and TLRs gene expressions in infected cell TLRs/MyD88 signal transduction pathway will be studied. The molecular mechanism and target of action on anti-Toxoplasm gondii of polysaccharide from Inonotus obliquus will be further verified and determined by adding TLRs/MyD88 signal molecule inhibitors. The effects of polysaccharide from Inonotus obliquus on mice infected by Toxoplasm gondii will also be studied by in vivo experiment. It will provide theoretical foundation and technological support for research and development of polysaccharide from Inonotus obliquus as a new anti-Toxoplasmosis drug, and has also certain significance for development and utilization of Inonotus obliquus in Changbai mountain area.

弓形虫病是由刚第弓形虫寄生于多种动物及人的有核细胞中引起的一种世界范围内广泛流行的人畜共患原虫病，严重危害着人类健康和畜牧业发展。本研究是在利用KEGG数据库和DAVID生物信息学分析工具对桦褐孔菌多糖治疗前后小鼠差异基因生物学通路进行分析，发现其可调控Toll样受体信号途径前期工作基础上，通过弓形虫感染宿主细胞体外研究桦褐孔菌多糖对感染的细胞TLRs/MyD88信号转导通路中关键蛋白表达和磷酸化水平、核转录因子AP-1和NF-κB活性、MyD88和TLRs等基因表达的影响；通过加入TLRs/MyD88信号通路分子抑制剂，进一步验证和确定桦褐孔菌多糖抗弓形虫分子机制和作用靶标；并通过弓形虫感染小鼠体内研究桦褐孔菌多糖对TLRs/MyD88信号转导通路的干预。这将为桦褐孔菌多糖作为抗弓形虫病新药的研发提供充分的理论依据和技术支撑，同时对长白山地区桦褐孔菌资源的开发和利用具有重要意义。

弓形虫病是由刚第弓形虫寄生于多种动物及人的有核细胞中引起的一种世界范围内广泛流行的人畜共患原虫病，严重危害着人类健康和畜牧业发展。本研究是在利用KEGG数据库和DAVID生物信息学分析工具对桦褐孔菌多糖治疗前后小鼠差异基因生物学通路进行分析，发现其可调控Toll样受体信号途径前期工作基础上，通过弓形虫感染宿主细胞体外研究了桦褐孔菌多糖对感染的细胞TLRs/MyD88信号转导通路关键蛋白表达和磷酸化水平、核转录因子AP-1和NF-κB活性、MyD88和TLRs等基因表达的影响，确定了桦褐孔菌多糖体外抗弓形虫分子机制和作用靶标；并通过弓形虫感染小鼠体内研究了桦褐孔菌多糖的保护作用及对信号转导通路的干预。研究表明：桦褐孔菌多糖体外通过调节弓形虫感染的小鼠RAW264.7巨噬细胞和小鼠脾淋巴细胞中TLR2/TLR4-NF-κB/MAPKs信号通路，影响细胞因子和趋化因子的分泌表达而抑制弓形虫感染所致的过度炎症反应，从而发挥其体外抗弓形虫作用；桦褐孔菌多糖体内通过调节TLRs/NF-κB和Nrf2/HO-1信号通路保护弓形虫感染所致的小鼠肝损伤，通过调节Th17/Treg平衡保护弓形虫感染所致孕鼠胎盘损伤从而改善孕鼠的妊娠结局，通过调节睾酮相关合成基因和凋亡蛋白保护弓形虫感染所致雄性小鼠生殖障碍从而提高生殖能力。这为桦褐孔菌多糖作为抗弓形虫病新药的研发提供了充分的理论依据和技术支撑，同时对长白山地区桦褐孔菌资源的开发和利用具有重要意义。