Toll样受体2和Toll样受体4及其信号通路在原发性痛风性关节炎发病机制中作用的研究

The role of TLR2, TLR4 and its signal pathway in gouty arthritis

目的探讨Toll样受体（TLR）2、TLR4及其信号通路在痛风炎症反应中的作用。方法应用实时荧光定量聚合酶链反应（RT-qPCR）法检测痛风性关节炎急性组32例、痛风性关节炎非急性组20例及健康对照组（健康体检者）32名外周血单个核细胞（PBMCs）TLR2mRNA、TLR4mRNA水平，Westem—blot检测上述3组各8例PBMCs核蛋白核因子-κBp65，酶联免疫吸附试验（ELISA）检测3组血浆白细胞介素（IL）-1β含量；并将上述各指标水平与痛风患者及健康体检者血尿酸水平进行相关性分析。多组间比较采用单因素方差分析，两两比较采用q检验。结果TLR4mRNA、核因子-κBD65、血浆IL-1β及血尿酸水平在痛风性关节炎急性组[（5．0±1．2）、（7．11±0．18）、（283±83）pg／ml、（585±123）μmol／L]和非急性组[（2．3±0．4）、（0．63±0．06）、（134±29）pg／ml，（493±107）pomol／L]均显著高于健康对照组[（1．1±0．6）、（0．52±0．12）、（97±17）pg／ml，（326±65）μmol／L]（P均〈0．01），痛风性关节炎急性组高于非急性组（P均〈0．01）；TLR2mRNA在3组的表达差异无统计学意义（P〉0．05）。痛风患者TLR4mRNA和IL-1β水平与血浆尿酸水平呈正相关（忙0．876，0．779；P均〈O．05），而TLR2mRNA和IL-1β水平与血浆尿酸水平无相关性（P均〉0．05）。健康体检者TLR4、TLR2mRNA与血尿酸、IL-1β水平均无相关性（P均〉0．05）。结论原发性痛风性关节炎可通过胞膜型模式识别受体激活固有性免疫应答，TLR4-核因子-κB—IL-1β信号通路参与了痛风免疫及炎症反应调节，痛风患者体内尿酸盐晶体与TLR4及其信号通路激活有关。

Objective The roles of TLRs and their signal pathway in gouty arthritis （GA） were explored. Methods TLR2 and TLR4 mRNA was measured using real-time quantitative polymerase chain reaction （RT-PCR） in PBMCs, IL-1β level was detected using ELISA in plasma, and NF-κB p65 protein level in PBMCs was measured using Western blot. Level of TLR2 mRNA, TLR4 mRNA, IL-1β, NF-κB p65 protein was compared among acute GA, non-acute GA and healthy controls. Correlation between TLR2 mRNA, TLR4 mRNA and serum uric acid, IL-1β level in GA patients was analyzed. One-way ANOVA was used to analyze data between multiple groups and q-test was used for two-two comparison. Spearman＇ s analysis was applied for correlation analysis. Results The expression of TLR4 mRNA, NF-κB p65 protein, IL-1β and serum uric acid level in patients with acute GA [（5.0±1.2）, （7.11±0.18）, （283±83） pg/ml, （ 585± 123 ） μmol/L ] was significantly increased compared to non-acute GA [ （ 2.3±0.4 ）, （0.63±0.06）, （ 134± 29） pg/ml, （493±107）txmol/L] and healthy controls （1.1±0.6）, （0.52±0.12）, （97±17）pg/ml, （326±65） μmol/L]（P〈0.01, respectively）. Significant difference was also observed between non-acute GA patients and healthy controls （P〈0.05, respectively）. The level of TLR4 mRNA was positively correlated with uric acid and IL-1β level in GA patients （rs=0.876, 0.779; P〈0.05, respectively）. Conclusion Innate immunity are activated by membrane-type pattern recognition receptors in primary GA. TLR4-NF-κB p65-IL-1β signal transduction may participate in the inflammatory mechanisms of gout. Urate crystals in patients with gout may be involved in the activation of TLR4 and its signal pathway.