MCC950对脂多糖诱导的胰腺腺泡细胞炎症的保护作用及机制  

Protective effect and mechanism of MCC950 on lipopolysaccharide induced pancreatic acinar cell inflammation

作　　者：阿布力克木·吾拉音 买买提·依斯热依力 王永康 叶勒丹·马汉 克力木·阿不都热依木[1,3] 吴朝阳[1] Abulikemu Wulayin;Maimaiti Yisireyili;Wang Yongkang;Yeledan Mahan;Kelimu Abudureyimu;Wu Zhaoyang(Department of Emergency Trauma Surgery,People’s Hospital of Xinjiang Uygur Autonomous Region,Urumqi Xingjiang 830001,China;Graduate School of Xinjiang Medical University,Urumqi Xingjiang 830017,China;Department of Minimally Invasive,Hernia and Abdominal Wall Surgery,People’s Hospital of Xinjiang Uygur Autonomous Region,Urumqi Xingjiang 830001,China)

机构地区：[1]新疆维吾尔自治区人民医院急诊创伤外科,新疆乌鲁木齐830001 [2]新疆医科大学研究生学院,新疆乌鲁木齐830017 [3]新疆维吾尔自治区人民医院微创、疝和腹壁外科,新疆乌鲁木齐830001

出　　处：《遵义医科大学学报》2024年第12期1157-1163,共7页Journal of Zunyi Medical University

基　　金：新疆维吾尔自治区自然科学基金资助项目(NO:2022D01C107);新疆维吾尔自治区优秀博士后普通基金资助项目(NO:346798)。

摘　　要：目的 探讨NOD样受体蛋白-3(NLRP-3)炎性小体抑制剂MCC950阻断脂多糖诱导胰腺腺泡细胞(PACs)炎症和焦亡的作用及相关机制。方法 小鼠胰腺腺泡细胞(MPACs)细胞株传代培养分为空白对照(Control组)、脂多糖(LPS组)、LPS+MCC950干预(MCC950组)以及抗氧化剂N-乙酰半胱氨酸(NAC组)4组,培养48 h。实时逆转录聚合酶链反应(RT-PCR)和蛋白质印迹法(WB)检测NLRP-3信号通路NLRP-3、ASC、caspase-1、IL-1β、IL-18以及细胞焦亡通路caspase-11、GSDMD的mRNA和蛋白表达。采用酶联免疫吸附(ELISA)测定方法检测活性氧指标丙二醛(MDA)的含量,Hoechst33342染色检测细胞凋亡水平。结果 MCC950以及NAC抗氧化剂干预有效抑制LPS诱导MPACs的凋亡。与Control组相比,PACs细胞加入LPS后显著增加丙二醛(MDA)的水平,而MCC950以及抗氧化剂NAC的干预有效地抑制LPS诱导MPACs细胞丙二醛(MDA)的高表达(P<0.01)。MCC950以及抗氧化剂NAC的干预有效抑制了LPS诱导MPACs细胞的NLRP-3、ASC、caspase-1、IL-1β、IL-18、caspase-11、GSDMD mRNA和蛋白质的高表达(P<0.01)。结论 MCC950对LPS诱导的急性胰腺炎具有保护作用,这可能与抑制细胞焦亡、NLRP-3信号通路的mRNA和蛋白表达有关。Objective To explore the effect and related mechanism of MCC950,an inflammasome inhibitor of NLRP-3,on blocking lipopolysaccharide induced inflammation and pyroptosis in pancreatic acinar cells(PACs).Methods Mouse pancreatic acinar cells(MPACs)were subcultured and divided into four groups:blank control group(Control group),lipopolysaccharide(LPS)group,LPS+MCC950 intervention group(MCC950 group),and antioxidant N-acetylcysteine(NAC group).The cells were cultured for 48 h.Real time reverse transcription polymerase chain reaction(RT-PCR)and western blot(WB)experiments were performed to detect and analyze the mRNA and protein expression levels of the NLRP-3 signaling pathway NLRP-3,ASC,caspase-1,IL-1β,and IL-18 and the cell apoptosis pathway caspase-11,and GSDMD.Enzyme linked immunosorbent assay(ELISA)was used to detect the level of malondialdehyde(MDA),a reactive oxygen species index.Cell apoptosis was detected by Hoechst33342 staining.Results MCC950 and NAC antioxidant intervention effectively inhibited LPS induced apoptosis of MPACs.Compared with the control group,the addition of LPS significantly increased the level of MDA in MPACs cells,while the intervention of MCC950 and antioxidant NAC effectively inhibited LPS induced high expression of MDA in MPACs(P<0.01).The intervention of MCC950 and antioxidant NAC effectively inhibited the high expression of mRNA and protein NLRP-3,ASC,caspase-1,IL-1β,IL-18,caspase-11,and GSDMD in LPS induced MPACs cells(P<0.01).Conclusion MCC950 has a protective effect on LPS induced acute pancreatitis,which may be related to the inhibition of expression of mRNA and protein related to the cell apoptosis,and NLRP-3 signaling pathway.

关 键 词：胰腺腺泡细胞 NOD样受体蛋白-3 脂多糖 MCC950 焦亡 

分 类 号：R576[医药卫生—消化系统]