从天然免疫信号TIR接头蛋白多态性表达负反馈角度解析清热解毒中药的多靶点抗炎效应

The TLR signaling of innate immune is the first line of defense of the host against pathogens, but excessive activation leads to systemic inflammatory response syndrome, therefore, to find molecular targets and effective drugs to balance innate immune is very important. TIR adapter proteins are the molecules of TLR signal transduction, it connects TLRs and kinases to decided to the direction of innate immune signal and affect the outcome. Previous studies found that these TIR adaptor proteins could express in polymorphism, resulting in producing negative variants, which balance the host defense and inflammation. No doubt, the negative feedback of TIR adapter protein variants will become a new target for the prevention and control of the innate immune imbalance. Traditional Chinese traditional medicines of heat-clearing and detoxicating have immunomodulatory and anti-inflammatory activities, although our previous studies of the NSFC found that these drug can interfere with TLR signaling, but its material foundation and multi-target effects lack of a reasonable explain base on modern medicine. To this end, we use the methods of molecular biology, bioinformatics and immunology to determine the endogenous mechanisms of balanced innate immune from the adapter protein polymorphism expression, further, use pharmacological methods to explore the substance basis and molecular mechanisms of Traditional medicines of heat-clearing and detoxicating base on the target of negative feedback of TIR adapter protein variants. In conclusion, our study will provide important experimental evidences to uncover the innate immune regulatory mechanism and relative drugs research, but also to provide new ideas for the modernization of traditional Chinese medicine.

天然免疫TLR信号是机体防御病原体第一道防线，但过度活化导致全身炎症反应综合症，因此，寻找其平衡的分子靶标和有效药物就显得尤其重要。TIR接头蛋白是TLR信号转导分子，它连接受体和激酶，决定天然免疫的信号流向，影响其结局。研究发现，这些TLR信号"二传手"可发生多态性表达，产生负性异构体，平衡机体防御和炎症。无疑，以TIR接头蛋白异构体为基础的负反馈将成为防控天然免疫失衡的新靶标。传统清热解毒中药具有免疫调节和抗炎活性，尽管先前NSFC研究发现其能干预TLR信号，但对它的物质基础和多靶点效应缺乏合理的现代医学阐释。为此我们用分子生物学、生物信息学和免疫学的方法，从接头蛋白多态性表达负反馈信号中确证平衡天然免疫的内源性机制，进一步以此为靶点用药理学方法解析清热解毒中药的物质基础和分子机制。总之，本课题为揭开天然免疫调控机制和相关药物研究提供重要的实验依据，同时也为传统中药现代化提供新的思路。

本课题是基于天然免疫TLR信号失调引起失控性炎症的病理生理机制并结合清热解毒中药临床应用，从TIR接头蛋白多态性表达调控角度探讨清热解毒中药的物质基础和多靶点抗炎作用的分子实质。通过课题组成员四年的相互协作和深入细致的工作，在天然免疫介导失控性炎症的发生机制和防治策略及清热解毒中药的物质基础和作用靶点研究方面取得一些重要结果：1）成功建立8种天然免疫TLR信号失调相关的急/慢性有菌或无菌炎症动物和细胞模型，并证实TLR信号通路激活和TIR接头蛋白多态性表达参与炎症的发生发展过程；2）通过评价5种经典清热解毒中药在不同急慢性失控性炎症模型中的作用，发现这5种清热解毒中药的抗炎作用与不同炎症模型类型（无菌或有菌）和急慢性特点密切相关； 2）证实其中2种清热解毒中药方剂和其包含的12种关键单体成分是通过调控TLR信号通路激活发挥抗炎效应；3）阐明HAT/HDAC-SRp/hnRNP-TIR adaptor信号机制调控TLR正负信号通路平衡，参与天然免疫TLR失调介导的重要脏器急慢性炎症损伤; 4) 从清热解毒中药方剂中筛选和证实了4种通过作用于SRp40或hnRNPA1影响TIR接头蛋白MyD88/TRIF/TRAM多态性表达，并有效抑制炎症反应，保护重要脏器损伤的单体化合物；5）通过纳米技术和化学修饰制备和改造清热解毒中药所包括的2种关键单体化合物，并证实其显著增强其对TLR相关炎症模型的抗炎活性。总之，通过本课题的研究，一方面从整体、细胞和分子水平阐明了炎症过程中TIR 接头蛋白多态性表达调控的关键反式剪接因子和信号调控的分子机制，这一发现必将揭开了以此为作用靶点干预天然免疫失控性炎症新的药物开发序幕；另一方面，通过此靶点筛选和评价了不同经典清热解毒中药的抗炎效应，并证实其多靶点抗炎的物质基础和分子靶标，为中药及其单体临床应用提供理论依据，也为传统中药现代化提供新的思路。