腺嘌呤核苷三磷酸-嘌呤受体P2X配体门控离子通道7-白细胞介素-1β通路参与痛风性关节炎发病的初步研究

The role of adenosine triphosphate-P2X receptor ligand gated ion channel 7-interleukin-1beta pathway in the pathogenesis of gouty arthritis

目的 探讨腺嘌呤核苷三磷酸-嘌呤受体P2X配体门控离子通道7-IL-β（ATP-P2X7R-IL-1β）通路在调控痛风性关节炎发病中的作用.方法 分离痛风与高尿酸血症患者外周血白细胞,分别用尿酸钠盐（MSU）和MSU＋ATP（5＇-三磷酸腺苷二钠盐水合物）刺激细胞培养,ELISA检测培养上清液中IL-1β浓度;流式细胞术检测外周血CD14＋P2X7R＋细胞.统计学处理采用t检验、非参数检验、Pearson相关分析.结果 ①单独MSU刺激时,痛风和高尿酸血症患者细胞培养液中IL-1β 的浓度[（50.9±1.5）pg/ml,（6.5±7.1）pg/ml]差异无统计学意义（Z=-1.088,P＞0.05）;用MSU＋ATP共同刺激白细胞时,痛风患者细胞培养液中 IL-1β 的浓度[（148±218） pg/ml]显著高于高尿酸血症患者[（24 ±39） pg/ml]（Z=-2.177,P＜0.05）.②高尿酸血症患者中,MSU刺激组[（6±7） pg/ml]和MSU＋ATP共同刺激组[（24±39） pg/ml]细胞培养液 中 IL-1β浓度差异无统计学意义（Z=-1.514,P＞0.05）;痛风患者中MSU＋ATP共同刺激组IL-1β浓度[（148± 218） pg/ml],显著高于MSU刺激组[（50.9±1.5） pg/ml],差异有统计学意义（Z=-2.704,P＜0.05）.③痛风患者外周血中CD14＋P2X7R＋细胞的百分比显著高于高尿酸血症患者[（1.6±0.5）％,（1.0±0.5）％],差异有统计学意义（t=-3.567,P＜0.05）.④痛风和高尿酸血症患者CD14＋P2X7R＋细胞的百分比与细胞培养液IL-1β浓度相关性均无统计学意义（r=-0.504,P＞0.05;r=-0.379,P＞0.05）.结论 ATP参与了痛风的发病,可能是引起急性痛风性关节炎发病的重要信号;其受体P2X7R的功能状态可能是决定痛风能否发病的重要因素.

Objective To investigate the role of adenosine triphosphate-P2X receptor ligand gated ion channel 7-interleukin-1beta （ATP-P2X7R-IL-1β） in gouty arthritis.Methods Peripheral white blood cells from patients with hyperuricemia or gout were cultured with monosodium urate crystals （MSU） or MSU＋ATP for 24 hours.IL-1β in supernatant was examined with enzyme linked immunosorbent assav （ELISA）.CD14＋ P2X7R＋ cells were detected using flow cytometry.T-test,non-parametric test,Pearson correlation analysis were used for statistical analysis.Results ① After stimulated by MSU alone,there was no difference in the concentration of IL-1β between the gout and hyperuricemia groups [（50.9±1.5） pg/ml,（6.5±7.1） pg/ml,Z=-1.088,P〉0.05].However,with the stimulation by MSU＋ATP,gout patients had higher concentration of IL-1β compared to the hyperuricemia patients [（148±218） pg/ml,（24±39） pg/ml,Z=-2.177,P〈0.05].② In hyperuricemia group,the concentrations of IL-1β was not different between MSU and MSU＋ATP stimulation [（6±7） pg/ml, （24±39） pg/ml,Z=-1.514,P〉0.05],while in gout group,the concentration was significantly higher after stimulated by MSU＋ATP than MSU [（50.9 ±1.5） pg/ml,（148 ±218） pg/ml,Z=-2.704,P〈0.05],③ The frequency of CD14＋P2X7R＋ cells in the peripheral blood of patients with gout was significantly higher than that of patients with hyperuricemia [（1 6 ±0.5）%,（1.0 ±0.5）%,t=-3.567,P〈0.05].④ No correlation was found between IL-1β concentration in supernatant and the frequency of CD14＋P2X7R＋ cells （r=-0.504, P=0.095：r=-0.379,P〉0.05）.Conclusion ATP is involved in the pathogenesis of gout,and is an important signal for the onset of acute gouty arthritis.Functional state of the receptor P2X7R may be an important factor to determine whether there will be gout attack.