Toll样受体炎性通路中相关基因功能区SNPs与痛风的关联研究及其民族异质性

Companying With lifestyle and dietary structure change, the incidence of gout increases year by year, and present the trend of youngness, the harm of gout caused global attaches. It's pathogenesity mainly because urate crystal(MSU) deposition and caused the inflammatory response by the innate immune system. The latest research shows MSU are recognized by Toll-like receptors (TLRs) and NALP3 inflammatory body,Uric acid salt crystals (MSU) deposition lead to inflammation maybe the mainly reason for gout,Toll-like receptors are the body's pattern recognition key receptors of MSU , MSU can induce the TLR2 and TLR4 activity of Toll -like receptor body inflammatory signaling pathway which in the macrophage and neutrophil cell surface,than activating transcription factor NF-κB and promote NALP3 inflammasome activity , eventually releasing a large number of inflammatory cytokines IL-1β, acting on synovial cell inflammation caused by gout stage linking reaction. This study intends to select those candidate genes which maybe will affected the expression of gout inflammation and inflammatory reaction resistance in Toll-like receptor -inflammatory pathway .Detect candidate gene promoter region SNPs in Han and Uygur male acute gout patients and control subjects, screening ifthese positive SNP can influnence the promoter funtional activity and protein expression of these genes, in order to establish Uygur and Han ethnic gout inflammation -related genes SNP database ,to clarify the signaling pathway of pro-inflammatory cytokines and inhibit inflammatory cytokine gene mutation in gout, to study the pathogenesis of gout in different ethnic, do basic research for gout pathogenesis and drug development. finaly activate the NF-κB release inflammatory cause gout cascaded inflammation reaction. This project is proposed based on our previous studies, study TLR2 and TLR4 receptor gene promoter region SNPs site and its influence in different ethnic gout patient and control; Analysis the variation rules, explore the inflammatory molecular mechanism and ethnic diversity of gout. In future we can found the Feasible prevention and control methods ,help for drug research and development to open up new research ideas for gout.

目前认为痛风急性发作与尿酸盐晶体(MSU)沉积导致的炎症反应有关，而Toll样受体是机体识别MSU的关键模式受体，MSU可诱导巨噬细胞和中性粒细胞膜表面Toll样受体炎性信号通路中关键受体TLR2和TLR4活性，活化转录因子NF-κB和促进NALP3 炎性体活性,最终释放大量炎性因子IL-1β，作用于关节滑膜细胞引发痛风性炎症的级联反应。本研究拟选择Toll样受体炎性通路上与痛风炎症反应最为密切的候选基因，通过分子生物学技术检测维汉男性急性痛风患者和对照人群候选基因启动子区SNPs，筛选出可能影响靶基因表达的SNP位点，观察阳性SNP位点对候选基因启动子活性的影响和蛋白表达，以期建立维汉民族痛风炎症反应相关基因的SNP数据库，阐明该信号通路基因突变在痛风炎症级联反应中的作用机制，寻找其变化规律，探索不同民族痛风发病机制研究的思路与途径，为开展痛风发病机制研究和药物研制奠定基础。

痛风急性发作与尿酸盐晶体沉积导致的炎症反应有关，而Toll样受体是机体识别MSU的关键模式受体，MSU可诱导巨噬细胞和中性粒细胞膜表面Toll样受体炎性信号通路中关键受体TLR2和TLR4活性，活化转录因子NF–κB和促进NALP3 炎性体活性,最终释放大量炎性因子IL-1β，作用于关节滑膜细胞引发痛风性炎症的级联反应。本研究选择Toll样受体炎性通路上与痛风炎症反应最为密切的候选基因TLR2、TLR4、NALP3等基因，通过分子生物学技术检测维汉男性急性痛风患者和对照人群候选基因启动子区SNPs，对各位点的基因型进行深入分析，探讨不同基因型下对痛风的影响，并在SNP多态性的基础上，选择有意义的位点进行蛋白功能验证。本次研究共选择了Toll样受体上相关基因的27个位点，所得结果在SNP基因分型上TLR2与痛风的发生均无统计学意义（P＞0.05），TLR4基因与痛风的发生均无统计学意义（P＞0.05），NALP3基因rs3806268位点不同分型下患痛风的差异有统计学意义（P＜0.05），IL-IB基因rs1143634位点不同分型下患痛风的差异有统计学意义（P＜0.05），其余分型无统计学意义，而对等位基因进行一步分析发现，每个位点均有有统计学意义的位点，可选取基因分型和等位基因有意义的位点进行蛋白验证的探究。本研究通过长期密切的实施，结果发现Toll样受体正如我们猜测的，在痛风疾病的发生发展中扮演着重要角色，今后可进一步深入探讨Toll样炎性受体通路对代谢性疾病的影响。