摘要
目的:探讨栀子苷对脂多糖(LPS)诱导的小鼠巨噬细胞RAW264.7 Caspase-1细胞焦亡经典信号通路的干预作用。方法:LPS诱导RAW264.7细胞构建炎症模型,以地塞米松为阳性对照,20、40、80μmol/L栀子苷干预细胞24 h。乳酸脱氢酶(LDH)细胞释放法检测细胞LDH释放;碘化丙啶(PI)荧光染色观察细胞存活情况;检测细胞上清中炎症因子IL-18、IL-1β、TNF-α和IL-6分泌水平;Western blot检测NLRP3、Caspase-1、GSDMD、IL-18和IL-1β表达。结果:与正常对照组相比,模型组RAW264.7细胞中LDH释放量明显升高(P<0.01),细胞形态学观察到有大量细胞被PI染色,细胞上清中IL-1β、IL-18、TNF-α和IL-6分泌水平显著升高(P<0.01);NLRP3、Caspase-1、GSDMD、IL-18和IL-1β蛋白表达显著升高(P<0.01)。与模型组相比,栀子苷各组和地塞米松组LDH释放减少(P<0.01),有效抑制PI染色,细胞上清中炎症因子IL-1β、IL-18、TNF-α和IL-6分泌水平降低(P<0.05);NLRP3、Caspase-1、GSDMD、IL-1β和IL-18表达降低(P<0.05)。结论:栀子苷可能通过抑制Caspase-1细胞焦亡信号通路激活减轻炎症反应。
Objective:To investigate effects of Geniposide on lipopolysaccharide(LPS)-induced mouse macrophages RAW264.7 in classical signaling pathway of Caspase-1 cell pyroptosis.Methods:LPS to induce RAW264.7 cells to constitute inflammation model.Dexamethasone was used as a positive control,and Geniposide at concentrations of 20,40,and 80μmol/L were used to intervene cells for 24 h.Lactate dehydrogenase(LDH)cell release method was used to detect release of cell LDH;propidium iodide(PI)fluorescent staining to observe cell survival;inflammatory factors IL-18,IL-1β,TNF-αand IL-6 secretion levels in cell supernatant were detected;expressions of NLRP3,Caspase-1,GSDMD,IL-18 and IL-1βwere detected by Western blot.Results:Compared with normal control group,release of LDH in RAW264.7 cells of model group was significantly increased(P<0.01);a large number of cells were stained by PI in cell morphology;cell supernatant secretion levels of IL-1β,IL-18,TNF-αand IL-6 were significantly increased(P<0.01);protein expressions of NLRP3,Caspase-1,GSDMD,IL-18 and IL-1βwere significantly increased(P<0.01).Compared with model group,LDH release was reduced in Geniposide groups and Dexamethasone group(P<0.01);PI staining were effectively inhibited,inflammatory factors IL-1β,IL-18,TNF-αand IL-6 secretion levels in cell supernatant were reduced(P<0.05);expressions of NLRP3,Caspase-1,GSDMD,IL-1βand IL-18 were decreased(P<0.05).Conclusion:Geniposide may reduce inflammatory response by inhibiting activation of Caspase-1 cell pyroptosis signaling pathway.
作者
游丽娇
杨小芳
耿欢
孟佳磊
马宇慧
雷鸣(指导)
王韫瑾
YOU Lijiao;YANG Xiaofang;GENG Huan;MENG Jialei;MA Yuhui;LEI Ming;WANG Yunjin(Intensive Care Unit,Seventh People's Hospital Affiliated to Shanghai University of TCM,Shanghai 200137,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2022年第13期1574-1578,共5页
Chinese Journal of Immunology
基金
国家自然科学基金面上项目(81973649)
上海市卫生健康委华东片区及市级中医专科专病联盟建设项目[ZY(2021-2023)-0302]
浦东新区卫生系统领先人才培养计划(pwr12019-02)项目
上海中医药大学预算内项目(2020HL189)
上海中医药大学附属第七人民医院人才培养计划(XX2022-12)资助。
