Pannexin-1 hemi-channels mediate pyroptosis on peripheral blood monocytes in patients with essential hypertension via regulating NLRP3/Caspase-1 pathway  

作　　者：ZHU Jian YANG Yan XU Shu-nan GAO Na KANG Pin-fang WANG Hong-ju 

机构地区：[1]Department of Cardiology the First Affiliated Hospital of Bengbu Medical College,Bengbu 233004,China [2]Department of Medical Oncology the First Affiliated Hospital of Bengbu Medical College,Bengbu 233004,China

出　　处：《Journal of Hainan Medical University》2023年第10期17-23,共7页海南医学院学报（英文版）

基　　金：National Natural Science Foundation of China(No.81970313);Natural Science Foundation of Anhui Province(No.2008085MH238);512 Talent Cultivation Plan of Bengbu Medical College(No.by51201321);Key Program of Natural Science Foundation of Bengbu Medical College(No.2020byzd109)。

摘　　要：Objective:To explore the role of NLRP3/Caspase-1-mediated pyroptosis on peripheral blood monocytes in patients with essential hypertension(EH),and its regulation by pannexin-1(Panx-1)hemi-channels.Methods:The peripheral blood of EH patients and healthy subjects was collected,peripheral plasma of the two groups was subsequently separated,and IL-1βcontent in peripheral plasma was further measured by ELISA.In the meantime,the monocytes from peripheral blood were sorted by immunomagnetic beads,and mRNA and protein expression of Panx-1,NLRP3 inflammasome related molecules(NLRP3,ASC,Caspase-1),downstream effector IL-1β,and pyroptosis-related protein GSDMD,were analyzed by RT-qPCR and Western blot,respectively.Subsequently,human primary monocytes in two group were cultured in vitro.Immunofluorescence assay was performed to detect the expression and location of Panx-1 on monocytes.Finally,the cultured monocytes from EH patients were exposed to NLRP3 inflammasome activator lipopolysaccharide(LPS),and also to LPS pretreated with Panx-1 hemi-channel inhibitor probenecid or specific Panx-1 siRNA,followed by collection of the cell culture supernatant and monocytes.The cell viability of each group was detected by CCK-8 assay,IL-1βcontent in the culture supernatant was analyzed by ELISA,and the expression of target proteins on monocytes was determined by Western blot assay.Results:Compared with healthy subjects,EH patients showed higher IL-1βcontent in peripheral plasma,and increased expression in both mRNA and protein levels for Panx-1,NLRP3 inflammasome related molecules,IL-1βand GSDMD on peripheral monocytes.Furthermore,Panx-1 protein in EH patients was significantly higher than that in healthy subjects and localized on monocyte membrane as evidenced by immunofluorescence assay.In the cultured human primary monocytes from EH patients,the cell viability in each group showed no significant difference.LPS stimulation can induce NLRP3-dependent pyroptosis,by showing an increase in IL-1βcontent in cell culture supernatant a

关 键 词：Essential hypertension Pannexin-1 NLRP3 inflammsome IL-1β GSDMD 

分 类 号：R544.1[医药卫生—心血管疾病]