痛风性关节炎急性发作期血尿下降的机制研究  

Mechanism of hematuria decline in acute gout arthritis

作　　者：高大玉 张均雩 李倩[1] 顾冰洁[1] 王小琴[1] 袁海 沈敏宁[1] 陈兴国[1] GAO Da-yu;ZHANG Jun-yu;LI Qian;GU Bing-jie;WANG Xiao-qin;YUAN Hai;SHEN Min-ning;CHEN Xing-guo(Department of Rheumatology and Immunology,Nanjing First Hospital,Nanjing Medical University,Nanjing 210006,Jiangsu,China)

机构地区：[1]南京医科大学附属南京医院(南京市第一医院)风湿免疫科,南京210006

出　　处：《医学研究生学报》2022年第1期69-74,共6页Journal of Medical Postgraduates

基　　金：南京市医学科技发展资金(YKK19075)。

摘　　要：目的目前外泌体与痛风性关节炎的相关性研究鲜有报道。文中旨在探索痛风性关节炎急性发作期血尿酸下降的机制。方法选取并收集2021年1月至2021年6月南京市第一医院风湿免疫科就诊的痛风患者和健康体检人员的临床资料和血清标本,并按照临床特则分为急性发作组、慢性缓解组、健康对照组。提取血清外泌体,利用Western blot、透射电镜、粒经分析对其进行鉴定,比较3组之间的差异。构建尿酸盐阴离子转运体1(URAT1)的表达载体,转染HEK293细胞,使得HEK293细胞稳定表达目的基因URAT1,并检测鉴定。将提取痛风患者的外泌体与过表达URAT1的HEK293细胞共培养,再加入高浓度尿酸共培养,检测尿酸浓度变化。结果①不同时期的血清外泌体浓度(×10^(6)Particles/mL):在痛风性关节炎急性发作期患者中最高(5.02±0.19),慢性缓解期稍高(4.17±0.64),正常人中最低(2.7±1.03),差异有统计学意义(P<0.01)。②稳定表达URAT1的HEK293细胞与外泌体共培养后,急性发作期组URAT1的表达较未加外泌体组明显下降(P<0.01),慢性缓解期略有下降(P<0.05)。③外泌体处理后的HEK293 URAT1^(+)细胞与高浓度尿酸共培养后,未加外泌体组细胞对尿酸的摄取率为(6.03±0.62)×10^(-4),加入慢性缓解组为(5.49±0.79)×10^(-4),加入急性发作组为(2.15±0.43)×10^(-4)。加入急性发作期外泌体的细胞对尿酸的摄取率明显下降(P<0.01)。结论痛风性关节炎患者血清外泌体浓度与疾病活动有关。外泌体可能通过作用于肾小管上皮细胞上的URAT1,抑制其对尿酸的重吸收,进而使肾排泄尿酸增加,最终使急性发作期患者血尿酸浓度下降。Objective Few reports are seen about the correlationbetween exosome and gouty arthritis.This study aimed to explorehow serum uric acid(SUA)declines amid the acute attack of gouty arthritis.Methods Healthy individuals and patients with gout treated in the Department of Rheumatology and Immunology,Nanjing First Hospital from January 2021 to June 2021 participated in this study and their clinical data and serum specimen were collected.They were divided into three groups as per their clinical presentations:acute attack group,chronic remission group and healthy control group.Serum exosomes were extracted and identified by Western blot,transmission electron microscope and grain analysis,then the differences between the three groups were compared.An expression vector for the urate anion transporter 1(URAT1)was constructed and transfected into the HEK293 cells,leading to a stable expression of the target gene URAT1 in HEK293 cells.Uric acid uptake experiment:Exosomes extracted from gout patients were co-cultured with overexpression HEK293 URAT1 cells,adding high concentrations of uric acid,then detected the change of uric acid concentration.Results Serum exosome level(×10^(6) Particles/mL)was the highest in patients with acute gouty arthritis(5.02±0.19),slightly higher in patients with chronic remission(4.17±0.64),and lower in healthyindividuals(2.7±1.03).The difference was statistically significant(P<0.01).When exosomes were co-cultured with HEK293 cells that stably expressed URAT1,the expression of URAT1 in the acute attack group was significantly decreased compared with that in the non-exosomes group(P<0.01),and slightly decreased in the chronic remission group(P<0.05).After co-culture of exosome-treated HEK293 URAT1+cells with high concentrations of uric acid,the rate of uric acid uptake were(6.03±0.62)×10^(-4) in the group without exosomes,(5.49±0.79)×10^(-4) in the chronic remission group,(2.15±0.43)×10^(-4) in the acute attack group.The uptake rate of uric acid was significantly decreased in the cells wi

关 键 词：痛风性关节炎 血尿酸 外泌体 尿酸盐阴离子转运体1 尿酸重吸收 

分 类 号：R684.3[医药卫生—骨科学]