Caspase-11/GSDMD非经典细胞焦亡途径在蓖麻毒素诱导RAW264.7细胞炎症反应中的作用  

Role of Caspase-11/GSDMD non-classical pyroptosis pathway in ricin toxininduced inflammation in RAW264.7 cells

作　　者：宋苏丽 董明鑫 孙成彪 王燕[2] 林茹 许娜 刘文森[1,2] SONG Suli;DONG Mingxin;SUN Chengbiao;WANG Yan;LIN Ru;XU Na;LIU Wensen(School of Life Science and Technology,Changchun University of Science and Technology,Changchun 130012,China;Changchun Institute of Veterinary Medicine,Chinese Academy of Agricultural Sciences,Changchun 130122,China;Changchun Lyuyuan District Maternal and Child Health and Family Planning Service Center,Changchun 130062,China;Jilin Medical College,Jilin 132000,China)

机构地区：[1]长春理工大学生命科学技术学院,长春130012 [2]中国农业科学院长春兽医研究所,长春130122 [3]长春市绿园区妇幼保健计划生育服务中心,长春130062 [4]吉林医药学院,吉林132000

出　　处：《中国免疫学杂志》2025年第4期898-902,907,共6页Chinese Journal of Immunology

基　　金：病原微生物生物安全国家重点实验室开放基金资助项目(SKLPBS2130)。

摘　　要：目的:探讨Caspase-11/GSDMD非经典细胞焦亡途径在蓖麻毒素诱导小鼠单核巨噬细胞RAW264.7炎症反应中的作用机制。方法:以LPS+Nigericin诱导的细胞焦亡模型为阳性对照,40、80 ng/ml蓖麻毒素诱导细胞8 h。乳酸脱氢酶(LDH)细胞释放法检测细胞LDH释放;流式细胞术(AnnexinⅤ/PI双染法)检测细胞焦亡;RT-qPCR检测Caspase-11、GSDMD、IL-1β和IL-18基因表达;Western blot检测Caspase-11和GSDMD蛋白水平;ELISA检测细胞上清中炎症因子IL-1β、IL-18分泌水平。结果:与正常对照组相比,蓖麻毒素处理组RAW264.7细胞发生肿胀、细胞膜破裂,LDH释放量显著升高(P<0.001),细胞焦亡率显著升高(P<0.001),焦亡相关基因Caspase-11、GSDMD、IL-1β和IL-18表达显著升高(P<0.05),Caspase-11、GSDMD-N蛋白表达显著升高(P<0.05),细胞上清中炎症因子IL-1β、IL-18分泌水平显著升高(P<0.05)。蓖麻毒素处理组与LPS+Nigericin细胞焦亡阳性对照组结果一致。结论:蓖麻毒素通过Caspase-11/GSDMD非经典细胞焦亡途径诱导RAW264.7细胞焦亡,在促进细胞炎症反应中具有重要作用。Objective:To explore mechanism of Caspase-11/GSDMD non-classical cell pyroptosis pathway in inflammatory response of RAW264.7 monocyte macrophages induced by ricin toxin.Methods:Cell pyroptosis model induced by LPS+Nigericin was used as a positive control,and cells were induced with 40 ng/ml and 80 ng/ml of ricin toxin for 8 hours.Cell release of lactate dehydrogenase(LDH)was measured by LDH cell release assay;cell viability was assessed by flow cytometry(AnnexinⅤ/PI double staining);gene expressions of Caspase-11,GSDMD,IL-1βand IL-18 were measured by RT-qPCR;protein levels of Caspase-11 and GSDMD were measured by Western blot;secretion levels of inflammatory cytokines IL-1βand IL-18 in cell supernatant were measured by ELISA.Results:Compared with normal control group,RAW264.7 cells treated with ricin toxin showed swelling,cell membrane rupture,significantly increased LDH release(P<0.001),significantly increased pyroptosis rate(P<0.001),significantly increased expression of pyroptosis-related genes Caspase-11,GSDMD,IL-1βand IL-18(P<0.05),significantly increased protein expressions of Caspase-11 and GSDMD(P<0.05),and significantly increased secretion levels of IL-1βand IL-18 in cell supernatant(P<0.05).Results of ricin toxin treatment group were consistent with LPS+Nigericin cell pyroptosis positive control group.Conclu-sion:Ricin toxin may induce cell pyroptosis in RAW264.7 cells through Caspase-11/GSDMD non-classical pathway,thereby promoting inflammatory response.

关 键 词：蓖麻毒素 细胞焦亡 Caspase-11 GSDMD 炎症反应 

分 类 号：R392.1[医药卫生—免疫学]