急性胰腺炎患者血清外泌体在NLRP3炎症小体介导的细胞焦亡中的作用机制研究  

Study on Mechanism of Serum Exosomes in NLRP3 Inflammasome-Mediated Pyroptosis in Patients with Acute Pancreatitis

作　　者：顾慧媛 沈佳庆[1] 高欣[1] 许春芳[1] 石玉琪 Gu Huiyuan;Shen Jiaqing;Gao Xin;Xu Chunfang;Shi Yuqi(Department of Digestive System,The First Affiliated Hospital of Soochow University,Suzhou 215000,China)

机构地区：[1]苏州大学附属第一医院消化内科,江苏苏州215000

出　　处：《南开大学学报（自然科学版）》2023年第6期80-84,共5页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：苏州市科技计划项目(SKY2021038)。

摘　　要：为了探究急性胰腺炎血清外泌体对于NLRP3炎症小体介导的细胞焦亡的作用机制.首先选取2021年1月至2023年1月苏州大学附属第一医院收治的85名急性胰腺炎患者为研究对象,根据急性生理与慢性健康状况II(APACHE-II)评分分为轻症组45例和重症组40例,两组患者外周血中NLRP3表达采用蛋白质印记检验,IL-1β和IL-18浓度采用ELISA检验,表面分子CD63和TSG101的水平采用流式法检验.结果表明:与轻症组相比,重症组患者APACHE-II评分显著升高,外周血NLRP3蛋白表达、IL-1β和IL-18浓度、CD63和TSG101水平均上升,且NLRP3蛋白表达与急性胰腺炎患者的APACHE-II评分呈正相关.在细胞层面,以AR42J细胞作为对照组,取急性胰腺炎重症组患者外周血分离后的外泌体与AR42J细胞共培育作为外泌体共培育组,比较两组细胞的NLRP3、caspase-1、caspase-4、caspase-5、Cleaved-Caspase11、GSDMD-FL和GSDMD-N表达、细胞上清中IL-1β和IL-18浓度.结果表明:与对照组相比,外泌体共培育组细胞的NLRP3、caspase-1、caspase-4、caspase-5、Cleaved-Caspase11、GSDMD-FL和GSDMD-N表达均上调,且细胞上清中IL-1β和IL-18水平上升.综上所述,急性胰腺炎患者血清外泌体分泌水平与疾病严重程度相关,通过介导NLRP3炎症小体激活可促进细胞焦亡,为探究临床上急性胰腺炎的致病机制奠定分子基础.In order to investigate the mechanism of cell pyroptosis mediated by serum exosomes in acute pancreatitis and NLRP3 inflammasome,85 patients with acute pancreatitis admitted to the First Affiliated Hospital of Soochow University from January 2021 to January 2023 were selected as the research subjects.According to the Acute Physiology and Chronic Health Evaluation II(APACHE-II)score,they were divided into a mild group of 45 cases and a severe group of 40 cases.The expression of NLRP3 in peripheral blood of patients in both groups was examined using a protein imprinting assay,and the concentrations of IL-1βand IL-18 were measured using ELISA.The levels of surface molecules CD63 and TSG101 were examined using flow cytometry.The results showed that compared to the mild group,the APACHE-II score of the severe group was significantly increased.The peripheral blood expression of NLRP3 protein,concentrations of IL-1βand IL-18,and levels of CD63 and TSG101 all increased.Moreover,the expression of NLRP3 protein was positively correlated with the APACHE-II score of patients with acute pancreatitis.At the cellular level,using AR42J cells as the control group,exosomes isolated from the peripheral blood of severe acute pancreatitis patients were co-cultured with AR42J cells as the exosome co-culture group.The expressions of NLRP3,caspase-1,caspase-4,caspase-5,Cleaved-Caspase11,GSDMD-FL,and GSDMD-N,as well as the concentrations of IL-1βand IL-18 in the cell supernatant,were compared between the two groups.The results showed that compared to the control group,the expression of NLRP3,caspase-1,caspase-4,caspase-5,Cleaved-Caspase11,GSDMD-FL,and GSDMD-N in the exosome co-culture group cells were upregulated,and the levels of IL-1βand IL-18 in the cell supernatant increased.In conclusion,the secretion level of serum exosomes in patients with acute pancreatitis is related to the severity of the disease.In conclusion,the level of serum exosome secretion in patients with acute pancreatitis is correlated with the severity of the dis

关 键 词：急性胰腺炎 血清外泌体 NLRP3炎症小体 细胞焦亡 

分 类 号：R576[医药卫生—消化系统] R392[医药卫生—内科学]