细胞焦亡相关分子在原发性痛风性关节炎中的表达及其临床意义  

Expression of pyroptosis related molecules in primary gouty arthritis and its clinical value

作　　者：雷天意 余湘 谢泓源 郭建伟 王鹏 张曾 张全波[2] 青玉凤[1] Lei Tianyi;Yu Xiang;Xie Hongyuan;Guo Jianwei;Wang Peng;Zhang Zeng;Zhang Quanbo;Qing Yufeng(Department of Rheumatology,Center for Hyperuricemia and Gout,Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,China;Department of Geriatrics,Affiliated Hospital of North Sichuan Medical College,Nanchong 637000,China)

机构地区：[1]川北医学院附属医院风湿免疫科高尿酸血症和痛风研究中心,南充637000 [2]川北医学院附属医院老年科,南充637000

出　　处：《中华风湿病学杂志》2023年第11期756-765,共10页Chinese Journal of Rheumatology

基　　金：国家自然科学基金面上项目(81974250);四川省南充市科技项目(20SXQT0308,20SXCXTD0002);川北医学院附属医院科研项目(2022ZD003)。

摘　　要：目的探讨原发性痛风性关节炎患者PBMCs中细胞焦亡的分子机制,为治疗痛风提供新思路。方法采集30例急性期痛风(AG)患者(AG组)、30例间歇期痛风(IG)患者(IG组)和40名健康患者(HC组)的外周血样本及临床资料;实时荧光定量检测细胞焦亡相关分子,包括核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、半胱氨酸天冬氨酸蛋白酶-1/4/5(caspase-1/4/5)、消皮素A/B/C/D/E(GSDMA/B/C/D/E)的mRNA表达水平。蛋白质免疫印迹法检测了NLRP3、前体caspase-1(pro-caspase-1)、剪切的caspase-1(caspase-1+p10)、GSDMD、N段GSDMD(GSDMD-N)、前体IL-1β(pro-IL-1β)、成熟IL-1β(cleced IL-1β)。正态或近似正态分布的计量资料采用独立样本t检验或单因素方差分析(ANOVA),非正态分布的计量资料采用Mann-WhitneyU检验或Kruskal-WallisH检验,计数资料使用χ^(2)检验进行比较。正态分布的连续性变量间采用Pearson相关分析,非正态分布的连续性变量间采用Spearman相关分析。Logistic回归分析用于评估危险因素。结果①一般资料比较:AG和IG组之间的药物持有率(MPR)、BMI差异无统计学意义(χ^(2)=0.64,P=0.426;t=0.04,P=0.972),病程差异具有统计学意义([25.0(9.8,63.0)月,54.0(33.0,102.0)月,Z=2.01,P=0.044)]。实验室指标比较:AG和IG组之间的ESR差异具有统计学意义(t=5.24,P<0.001),肾小球滤过率(eGFR)、中性粒细胞(GR)、淋巴细胞(LY)、红细胞(RBC)、红细胞压积(HCT)、尿酸、肌酐、ALT、AST 3组差异之间具有统计学意义。②3组间caspase-1、GSDMC、GSDMD、GSDME、NLRP3 mRNA表达水平差异有统计学意义。其中,在AG、IG组中caspase-1[(1.55±0.62、1.58±0.62)、GSDMD(4.7±1.4、3.5±1.5)]、NLRP3[(2.63(2.03,4.10)、2.39(1.57,3.49)]mRNA表达水平高于HC组[1.24±0.59,P=0.037,P=0.023;1.16±0.71,P<0.001,P<0.001;1.16(0.52,2.34),P<0.001,P<0.001],在IG组中GSDMD(3.5±1.5)mRNA表达水平相比较AG组(4.7±1.4)降低(P<0.001),而在AG组GSDMC、GSDME[(0.57(0.33,0.78)、(0.32±0.Objective To explore the molecular mechanism of cell death of the peripheral blood mononuclear cells(PBMCs)of patients with primary gouty arthritis,and provide new idea for the treatment of gout.Methods Peripheral blood samples and clinical data were collected from 30 patients with acute gout(AG),30 patients with intermittent gout(IG)and 40 healthy controls(HC).Real-time fluorescence quantitative detection of cell apoptosis related molecules,including the mRNA expression level of nucleotide binding oligomerization domain like domain like receptor protein 3(NLRP3),cysteine aspartic proteinase-1/4/5(caspase-1/4/5),Gasdermin A/B/C/E.And NLRP3,precursor caspase-1(pro-caspase-1),clipped caspase-1(caspase-1+p10),Gasdermin D(GSDMD),N segment GSDMD(GSDMD-N),precursor IL-1β(pro IL-1β),mature IL-1β(clevated IL-1β)were detected by western blot.The measurement data of normal or approximate normal distribution were analyzed by independent sample t test or one-way variance analysis(ANOVA),the measurement data of non-normal distribution were analyzed by Mann-Whitney U test or Kruskal-Wallis H test,and the counting data was compared by Chi-square test.Pearson's correlation analysis was used for the continuous variables with normal distribution,and Spearman's correlation analysis was used for the continuous variables with non-normal distribution.The logistic regression analysis was used to assess risk factors.Results①There were no significant differences in MPR and BMI between AG and IG(χ^(2)=0.64,P=0.426;t=0.04,P=0.972),and there was significant difference in disease course[25.0(9.8,63.0),54.0(33.0,102.0)mouth,Z=2.01,P=0.044].Comparison of labora-tory parameters:there were statistical significant differences in ESR between AG and IG(t=5.24,P<0.001),eGFR,GR,LY,RBC,HCT,UA,Creatinin,ALT and AST.②In the three groups,the expression lev-els of caspase-1,GSDMC,GSDMD,GSDME,NLRP3 mRNA were statistically significantly different.In AG and IG groups,mRNA expression levels of caspase-1(1.55±0.62),(1.58±0.62),GSDMD(4.7±1.4),(3.5±

关 键 词：关节炎 痛风性 细胞焦亡 炎性体 半胱氨酸天冬氨酸蛋白酶 消皮素 

分 类 号：R589.7[医药卫生—内分泌]