摘要
目的:探讨草苁蓉多糖(BRPS)对脂多糖(LPS)联合三磷酸腺苷(ATP)诱导的小鼠J774A.1巨噬细胞炎症小体活化及细胞焦亡的抑制作用。方法:LPS联合ATP刺激J774A.1巨噬细胞,建立巨噬细胞核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体模型。用细胞增殖毒性试剂盒检测J774A.1细胞存活率,微板法测定细胞培养液中乳酸脱氢酶(LDH)的释放,免疫荧光染色法检测细胞膜损伤情况,酶联免疫吸附法检测细胞培养液中白细胞介素-1β(IL-1β)水平,免疫印迹法测定J774A.1细胞NLRP3、Caspase-1、凋亡相关斑点样蛋白(ASC)、IL-1β、消皮素D(GSDMD)以及核转录因子-κB(NF-κB)、丝裂原活化蛋白激酶(MAPK)蛋白的表达情况。结果:BRPS能增高LPS/ATP诱导的J774A.1巨噬细胞存活率,降低细胞培养液中LDH释放,减少细胞膜损伤,下调细胞NLRP3蛋白表达,上调Caspase-1前体蛋白水平,下调IL-1β以及GSDMD N端活性片段水平。此外,BRPS下调J774A.1巨噬细胞NF-κB核转位及其磷酸化水平,以及细胞外信号调节蛋白激酶、c-Jun氨基末端激酶、p38 MAPK磷酸化水平。结论:BRPS可抑制LPS/ATP诱导的小鼠J774A.1巨噬细胞炎症小体活化和焦亡,其作用可能与调控NF-κB和MAPK信号通路有关。
Objective:This study aimed to investigate the inhibitory effect of Boschniakia rossica polysaccharides(BRPS)on the activation of inflammasome and pyroptosis of J774A.1 macrophages induced by lipopolysaccharide(LPS)and adenosine triphosphate(ATP).Methods:The nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)inflammasome model was established by stimulating J774A.1 macrophages with LPS and ATP.Cell viability was detected using the cell counting kit-8,the release of lactate dehydrogenase(LDH)was tested with the microplate method,the cellular level of NLRP3 and cell membrane damage were detected using the immunofluorescence staining method,and the secretion of interleukin-1β(IL-1β)was measured by the enzyme-linked immunosorbent assay.The protein expressions of NLRP3,caspase-1,apoptosis associated speck-like protein containing caspase recruitment domains(ASC),IL-1β,gasdermin D(GSDMD),as well as the nuclear factors-κB(NF-κB)and mitogen activated protein kinase(MAPK)was determined with the western blotting method.Results:BRPS could increase the cell viability,reduce the release of LDH in the culture medium,alleviate the membrane damage of macrophages,down-regulate the protein expression of NLRP3,up-regulate the level of caspase-1 precursor,and down-regulate the levels of IL-1β and GSDMD N-terminal active fragment in the model group.In addition,BRPS suppressed the nuclear translocation and phosphorylation of NF-κB,as well as the phosphorylation of extracellular signal-regulated protein kinase,c-Jun amino terminal kinase,and p38 MAPK.Conclusion:BRPS inhibit LPS/ATP-induced activation of macrophage inflammasome and pyroptosis in mouse J774A.1 cells through regulating the NF-κB and MAPK signaling pathways.
作者
张天
徐慧
全吉淑
金爱花
ZHANG Tian;XU Hui;QUAN Jishu;JIN Aihua(Yanbian University Hospital,Yanji,Jilin 133000,China;Medical College of Yanbian University,Yanji,Jilin 133000,China)
出处
《食品与机械》
CSCD
北大核心
2024年第5期137-145,共9页
Food and Machinery
基金
国家自然科学地区科学基金项目(编号:82060113)。
关键词
草苁蓉
多糖
巨噬细胞
炎症小体
信号通路
Boschnikia rossica
polysaccharides
macrophages
inflammasome
signaling pathways
