Department of Hepatopathy, Hunan Provincial People's Hospital (the first affiliated hospital of Hunan NormalUniversity), Changsha, Hunan, China.
Iran J Immunol. 2022 Mar;19(1):4. doi: 10.22034/IJI.2022.90579.2016.
Pyroptosis is a programmed cell death related to caspase-1, accompanied by the secretion of pro-inflammatory cytokines.
To explore the effects of LPS on the P2X7R/NLRP3 pathway in macrophages, and hepatocytes pyroptosis in mice.
LPS was used to establish an animal model of the acute liver injury. The macrophage RAW264.7 was induced by LPS to establish a cell model. The P2X7R inhibitor A438079 and agonist BZATP were added. RAW264.7 was co-cultured with AML-12 cells. Pyroptosis and the ratio of CD11b+CD86+/CD11b+CD206+ were analyzed by flow cytometry. ELISA, WB, and qRT-PCR were applied to analyze factors involved in the P2X7R/NLRP3 pathway.
LPS induced liver damage in mice, promoted cell pyroptosis and increased the levels of IL-18, IL-1β, ALT, AST, and TBIL. P2X7R, GSDMD, and GSDMD-N expressions also increased in the LPS group. LPS induced macrophage activation in vivo. NLRP3, ASC, P2X7R, and caspase-1 expressions in vitro promoted. ELISA confirmed that the IL-1β and IL-18 levels repressed in the BZATP (P2X7R agonist) group, while the trend was opposite in the A438079 (P2X7R inhibitor) group. LPS activated the P2X7R/NLRP3 pathway in macrophages. After RAW264.7 was co-cultured with AML-12 cells, the pyroptosis of AML-12 cells promoted but the proliferation decreased in the BZATP group. GSDMD and GSDMD-N expressions promoted in the BZATP group, while the trend was opposite in the A438079 group.
LPS activated macrophages via P2X7R activation of NLRP3 and induced hepatocyte pyroptosis, which provided novel potential targets for the liver injury treatment.
细胞焦亡是一种与半胱天冬酶-1（caspase-1）相关的程序性细胞死亡，伴有促炎细胞因子的分泌。
探讨脂多糖（LPS）对巨噬细胞 P2X7R/NLRP3 通路及小鼠肝细胞焦亡的影响。
采用 LPS 建立急性肝损伤动物模型，诱导 RAW264.7 巨噬细胞建立细胞模型，加入 P2X7R 抑制剂 A438079 和激动剂 BZATP，与 AML-12 细胞共培养。通过流式细胞术分析细胞焦亡及 CD11b+CD86+/CD11b+CD206+的比例，采用 ELISA、WB、qRT-PCR 分析 P2X7R/NLRP3 通路相关因子。
LPS 诱导小鼠肝损伤，促进细胞焦亡，增加 IL-18、IL-1β、ALT、AST、TBIL 水平，LPS 组 P2X7R、GSDMD、GSDMD-N 表达也增加。LPS 诱导体内巨噬细胞活化，体外促进 NLRP3、ASC、P2X7R、caspase-1 表达，ELISA 证实 BZATP（P2X7R 激动剂）组 IL-1β、IL-18 水平下降，A438079（P2X7R 抑制剂）组呈相反趋势。LPS 激活巨噬细胞 P2X7R/NLRP3 通路，RAW264.7 与 AML-12 细胞共培养后，BZATP 组促进 AML-12 细胞焦亡，但增殖减少，BZATP 组 GSDMD、GSDMD-N 表达增加，A438079 组呈相反趋势。
LPS 通过激活 P2X7R 激活 NLRP3 激活巨噬细胞，诱导肝细胞焦亡，为肝损伤治疗提供了新的潜在靶点。
