自噬相关基因Beclin-1在间歇期痛风性关节炎患者外周血单个核细胞中的表达变化及意义  

Expression change and clinical significance of autophagy-related gene Beclin-1 in peripheral blood mononuclear cells of patients with intercritical gouty arthritis

作　　者：杨其彬[1] 何泳龙 青玉凤[1] 李玲琴[1] 谢文光 周京国 YANG Qibin;HE Yonglong;QING Yufeng;LI Lingqing;XIE Wenguang;ZHOU Jingguo(The Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,China)

机构地区：[1]川北医学院附属医院,四川南充637000 [2]成都医学院第一附属医院

出　　处：《山东医药》2018年第37期26-29,共4页Shandong Medical Journal

基　　金：四川省科技厅科研项目(2017JY0037);四川省教育厅科研项目(15ZB0202);四川省医学会科研项目(S16027);四川省卫生和计划生育委员会科研课题(17PJ059)

摘　　要：目的探讨自噬相关基因Beclin-1在间歇期痛风性关节炎(IGA)患者外周血单个核细胞(PBMC)中的表达变化及意义。方法采集40例间歇期IGA男性患者(IGA组)、80例体检健康男性(正常对照组)外周静脉血,分离PBMC并进行培养。用实时荧光定量PCR、Western blotting技术分别检测PBMC中Beclin-1 mRNA、蛋白表达。随机抽取5例IGA患者外周静脉血20 m L,分为未刺激组及尿酸盐晶体(MSU)刺激1、3、6 h组并提取其PBMC,用Western blotting技术检测PBMC中Beclin-1蛋白表达。随机抽取5例IGA患者、5例体检健康者外周静脉血20 m L,将血液标本均随机分为空白对照组、雷帕霉素组、自噬抑制剂组。空白对照组不做干预,其他各组均加入相应试剂进行干预并提取PBMC,用Western blotting技术检测PBMC中Beclin-1蛋白表达;用酶联免疫吸附试验测定干预后IGA患者血浆IL-1β水平。结果与正常对照组比较,IGA组PBMC中Beclin-1 mRNA、蛋白相对表达量低(P均<0. 05)。MSU刺激1、3 h组PBMC中Beclin-1蛋白相对表达量较未刺激组高(P均<0. 05),而MSU刺激6 h组Beclin-1蛋白相对表达量较未刺激组低(P <0. 05)。IGA患者PBMC中Beclin-1蛋白相对表达量比较:雷帕霉素组较空白对照组低(P <0. 01),且自噬抑制剂组最高(P均<0. 05),恢复至空白对照组水平;体检健康者PBMC中Beclin-1蛋白相对表达量比较:空白对照组、MSU组、雷帕霉素组比较差异无统计学意义(P均> 0. 05),而自噬抑制剂组较其他组低(P均<0. 05)。IGA患者血浆IL-1β水平比较,MSU、雷帕霉素组较空白对照组高(P均<0. 05),雷帕霉素组较MSU组低(P <0. 05),自噬抑制剂组较空白对照组、MSU组高(P均<0. 05),自噬抑制剂组之间比较差异无统计学意义(P均> 0. 05)。结论自噬相关基因Beclin-1在间歇期IGA患者外周血PBMC中呈低表达,其可能在IGA发展过程中参与自噬途径并最终被自噬溶酶体降解。Objective To explore expression change and clinical significance of autophagy-related gene Beclin-1 in the peripheral blood mononuclear cells(PBMCs)of patients with intercritical gouty arthritis(IGA).Methods The peripheral venous blood was collected from 40 IGA male patients(IGA group)and 80 healthy men(healthy control group)and then the PBMCs were isolated and cultured.Beclin-1 mRNA and protein expression in the PBMCs were detected by real-time fluorescence quantitative PCR and Western blotting.Twenty mL peripheral venous blood from 5 IGA patients was randomly selected,and was randomly divided into MSU stimulation group(1-,3-and 6-hour groups)and non-stimulation group,and the PBMCs were extracted.Beclin-1 protein expression in the PBMCs was detected by Western blotting.Twenty mL peripheral venous blood was obtained from 5 IGA patients and 5 healthy individuals randomly and was equivalently divided into the blank control group,rapamycin group,and autophagy inhibitor group.No intervention was performed in the blank control group,and corresponding reagents were added to the other groups for intervention and PBMCs were further extracted.Beclin-1 protein expression in the PBMCs was detected by Western blotting.The plasma IL-1βlevels in the IGA patients were also determined by enzyme-linked immunosorbent assay.Results Compared with the healthy control group,Beclin-1 mRNA and protein expression was relatively lower in the IGA group(P<0.05).Beclin-1 protein expression was higher in the PBMCs of MSU stimulation 1-hour and 3-hour groups than in the non-stimulation group(P<0.05),while Beclin-1 protein expression was lower in the MSU stimulation 6-hour group than in the non-stimulation group(P<0.05).Comparison of Beclin-1 protein expression in the PBMCs of IGA patients:compared with the blank control group,the Beclin-1 protein expression in the rapamycin group was lower(P<0.01),and was the highest in the autophagy inhibitor group(P<0.05),and its level was restored to that of the blank control group.However,there was no si

关 键 词：痛风性关节炎 间歇期 细胞自噬 Beclin-1基因 

分 类 号：R593.22[医药卫生—内科学]