【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。更多还原

【Abstract】 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.更多还原