双酚A暴露与肥胖患者脂肪组织NLRP3炎症小体激活的相关性

Correlation between bisphenol A level and activation of NLRP3 inflammasome in adipose tissue of obese patients

目的通过临床病例样本检测分析探讨双酚A(bisphenol A,BPA)暴露与肥胖患者脂肪组织NLRP3炎症小体激活的相关性。方法纳入代谢减重手术的肥胖患者102例,收集临床资料及人口统计学数据,并采集血、尿和手术切除的内脏脂肪组织标本。液相色谱-质谱联法(liquid chromatography mass spectrometry,LC-MS)检测尿BPA浓度(肌酐校正BPA,简称BPAcr),ELISA检测血清、脂肪组织炎性细胞因子白细胞介素(interleukin,IL)-1β和IL-18含量,免疫荧光法检测脂肪组织NLRP3表达,qRT-PCR和免疫印迹法检测脂肪组织NLRP3激活相关分子的mRNA和蛋白表达水平,分析尿BPAcr与血清和脂肪组织炎性因子水平,以及脂肪组织NLRP3激活的相关性。结果肥胖患者尿BPAcr与BMI、血清IL-1β、血清IL-18、脂肪组织炎性因子IL-1β和IL-18水平呈正相关(r_(s)值分别为0.784、0.852、0.737、0.509及0.471,均有P<0.001)。肥胖患者脂肪组织巨噬细胞(adipose tissue macrophages,ATMs)中NLRP3表达,NLRP3、ASC mRNA表达水平,以及IL-1β、caspase-1蛋白表达水平与尿BPAcr均相关(均有P<0.05)。结论BPA暴露可能通过激活脂肪组织ATMs中NLRP3炎症小体,上调促炎细胞因子的分泌,导致肥胖患者炎症状态。

Objective The correlation between bisphenol A(BPA)exposure and the activation of NLRP3 inflammasomes in adipose tissue of obese patients was explored through the detection and analysis of clinical case samples.Methods A total of 102 obese patients undergoing metabolic surgery were included,and clinical data,demographic data,blood,urine,and surgically removed visceral fat tissue specimens were collected.Liquid chromatography mass spectrometry(LC-MS)was used to detect urine BPA concentration(Creatinine corrected BPA,BPAcr for short),and ELISA was used to detect serum and adipose tissue inflammatory cytokines IL-1βand IL-18 content.Immunofluorescence was used to detect the expression of NLRP3 in adipose tissue,qRT-PCR and immunoblot were used to detect the mRNA and protein expression levels of NLRP3 activation related molecules in adipose tissue.The correlation between urinary BPAcr and inflammatory factors in serum and adipose tissue,as well as the activation of NLRP3 in adipose tissue were analyzed.Results Urine BPAcr in obese patients was significantly positively correlated with body mass index(BMI),serum IL-1β,serum IL-18 adipose tissue inflammatory factors IL-1βand IL-18 levels(r_(s)values were 0.784,0.852,0.737,0.509,0.471,all P<0.001).The expression of NLRP3,the expression levels of NLRR3,ASC mRNA,and the protein expression levels of IL-1βand caspase-1 in ATMs of obese patients were significantly correlated with urinary BPAcr(all P<0.05).Conclusion BPA exposure may activate the NLRR3 inflammasome in ATMs of adipose tissue and up-regulate the secretion of pro-inflammatory cytokines,leading to the inflammatory state of obese patients.