JNK抑制剂SP600125抑制白细胞介素1β诱导的βTC-6细胞焦亡  

JNK inhibitor SP600125 inhibits interleukin-1βinduced pyroptosis inβTC-6 cells

作　　者：麻文淼 刘腾丽 梁瑞[2] 刘娜[2] 王乐[2] 王树森 郑荣秀[1] Ma Wenmiao;Liu Tengli;Liang Rui;Liu Na;Wang Le;Wang Shusen;Zheng Rongxiu(Department of Pediatrics,General Hospital of Tianjin Medical University,Tianjin 300052,China;NHC Key Laboratory for Critical Care Medicine,Tianjin First Central Hospital,Nankai University,Tianjin 300192,China)

机构地区：[1]天津医科大学总医院儿科,天津300052 [2]南开大学附属第一中心医院,卫生健康委员会急救医学重点实验室,天津300192

出　　处：《中华内分泌代谢杂志》2023年第5期430-434,共5页Chinese Journal of Endocrinology and Metabolism

基　　金：国家重点研发计划课题(2020YFA0803704);国家自然科学基金(82070805、82100840、82100841);天津市"项目+团队"重点培养专项(XB202011);天津市科技计划项目(20JCQNJC01720、ZC20222);天津市医学重点学科建设项目(TJYXZDXK-068C)。

摘　　要：目的研究白细胞介素(IL)-1β是否参与细胞焦亡造成的小鼠胰岛β细胞系βTC-6细胞损伤,以及JNK抑制剂SP600125在抑制IL-1β诱导的βTC-6细胞焦亡中的作用。方法βTC-6细胞系及小鼠胰岛,给予IL-1β孵育48 h,同时给予JNK抑制剂SP600125或IL-1受体拮抗剂(IL-1Ra)干预,然后通过免疫荧光染色GSDMD和DAPI检测GSDMD表达并观察细胞焦亡形态,实时荧光定量PCR检测细胞焦亡标志基因Gsdmd,IL-1β,IL-18 mRNA表达水平;采用Annexin-V/7-AAD染色结合流式细胞术检测细胞凋亡。结果与对照组相比IL-1β处理组中βTC-6细胞焦亡小体显著增加,焦亡相关基因Gsdmd、IL-1β、IL-18 mRNA表达明显升高(P<0.05),且细胞凋亡增加;经IL-1Ra处理后可有效抑制IL-1β诱导的βTC-6细胞焦亡,提示IL-1β有效的诱导βTC-6细胞焦亡的发生。在IL-1β处理时给予SP600125干预,则抑制细胞焦亡标志物Gsdmd和IL-18 mRNA表达,并且降低细胞凋亡率,说明SP600125抑制IL-1β诱导的βTC-6细胞焦亡。结论细胞焦亡是IL-1β造成的βTC-6细胞损伤的机制之一,而JNK抑制剂SP600125可阻断IL-1β诱导的焦亡途径,具有潜在的抑制βTC-6细胞焦亡的作用。Objective To investigate whether interleukin(IL)-1βis involved in pyroptosis which leads to mouse isletβcell lineβTC-6 cell damage,and to explore the role of JNK inhibitor SP600125 in inhibiting IL-1βinducedβTC-6 cell pyroptosis.MethodsβTC-6 cell line and mouse islets were incubated with IL-1βfor 48 h or intervened with both JNK inhibitor SP600125 and IL-1R antagonist IL-1Ra,then GSDMD expression andβcell pyroptosis morphology were detected by immunofluorescence staining of GSDMD and DAPI.The expression levels of Gsdmd,IL-1βand IL-18 mRNAs were detected by real time fluorescence PCR,and apoptosis was examined by Annexin-V/7-AAD staining combined with flow cytometry.ResultsβTC-6 cell pyroptotic body was significantly increased in the IL-1βtreated group compared with the control group,and the expressions of pyroptosis related genes Gsdmd,IL-1β,and IL-18 mRNA were significantly higher(P<0.05),and apoptosis was increased,suggesting that IL-1βeffectively induced theβTC-6 cell pyroptosis,IL-1Ra prevented IL-1βinducedβTC-6 cell pyroptosis.In the presence of JNK inhibitor SP600125,IL-1βtreatment failed to induce the expressions of Gsdmd and IL-18 mRNA,markers of pyroptosis,and reduced the rate of apoptosis,indicating that SP600125 suppressed IL-1βinducedβTC-6 cell pyroptosis.Conclusion Pyroptosis is one of the mechanisms ofβTC-6 cell impairment caused by IL-1β,and SP600125,a JNK inhibitor,can block the IL-1βinduced pyroptosis pathway and has a potential role in inhibitingβTC-6 cell pyroptosis.

关 键 词：细胞焦亡 糖尿病 GSDMD 白细胞介素-1Β β细胞系 

分 类 号：R587.1[医药卫生—内分泌]