中药新药与临床药理2025，Vol.36Issue(2)：206-213,8.DOI:10.19378/j.issn.1003-9783.2025.02.006
左归降糖舒心方含药血浆对脂多糖诱导巨噬细胞损伤及焦亡的影响
Effect of Zuogui Jiangtang Shuxin Formula-containing Plasma on LPS-induced Macrophage Damage and Pyroptosis
摘要
Abstract
Objective To investigate the effect and mechanism of Zuogui Jiangtang Shuxin Formula-containing plasma on LPS-induced macrophage damage and pyroptosis based on Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)/NOD-like receptor thermal protein domain associated protein 3(NLRP3)pathway.Methods Fully differentiated J774A.1 cells were randomly divided into the blank control group(10%blank plasma),the model group(10 μg·mL-1 LPS+10%blank plasma),5%(10 μg·mL-1 LPS+5%Zuogui Jiangtang Shuxin Formula-containing plasma)and 10%(10 μg·mL-1 LPS+10%Zuogui Jiangtang Shuxin Formula-containing plasma)Zuogui Jiangtang Shuxin Formula-containing plasma group.Except for the blank control group,other groups were treated with 10 μg·mL-1 LPS to induce inflammatory injury and pyroptosis of macrophage.After the intervention,ELISA was used to detect concentration of tumor necrosis factor-alpha(TNF-α),interleukin 6(IL-6),interleukin 1 beta(IL-1β)and interleukin 18(IL-18)in macrophage supernatant.The co-expression of NOD-like receptor protein 3(NLRP3)and apoptotic spot-like protein(ASC)were detected by immunofluorescence assay.The expression of TLR4/NF-κB/NLRP3 signaling pathway and pyroptosis-related proteins were detected by Western Blot.Results Compared with the blank control group,the levels of TNF-α,IL-6,IL-1β and IL-18 in the supernatant of LPS-treated J774A.1 cells significantly increased(P<0.01).It was also found that the fluorescence intensity of NLRP3 and ASC in cells was significantly enhanced,and protein co-expression was significantly up-regulated.The protein co-expressions of TLR4,p-NF-κB p65,NLRP3,GSDMD-N and IL-1β were significantly up-regulated(P<0.01).Compared with the model group,the levels of TNF-α,IL-6,IL-1β and IL-18 in the supernatant of Zuogui Jiangtang Shuxin Formula-containing plasma-treated J774A.1 cells were obviously decreased(P<0.01).The fluorescence intensity of NLRP3 and ASC in cells was significantly reduced,and protein co-expression was significantly down-regulated.The protein co-expressions of TLR4,p-NF-κB p65,NLRP3,GSDMD-N and IL-1β were significantly down-regulated(P<0.01).Conclusion Zuogui Jiangtang Shuxin Formula-containing plasma can suppresses LPS-induced macrophage damage and inflammatory response,and its mechanism may be related to the regulation of TLR4/NF-κB/NLRP3 signaling pathway and inhibition of cellular pyroptosis and inflammatory factor expression.关键词
左归降糖舒心方含药血浆/J774A.1巨噬细胞/TLR4/NF-κB/NLRP3炎症通路/细胞焦亡/炎症因子Key words
Zuogui Jiangtang Shuxin Formula-containing plasma/J774A.1 cells/TLR4/NF-κB/NLRP3 signaling pathway/pyroptosis/inflammatory factor分类
医药卫生引用本文复制引用
杨金伟,周毅,肖凡,喻嵘..左归降糖舒心方含药血浆对脂多糖诱导巨噬细胞损伤及焦亡的影响[J].中药新药与临床药理,2025,36(2):206-213,8.基金项目
国家自然科学基金项目(82205077) （82205077）
湖南省自然科学基金项目(2022JJ40315) （2022JJ40315）
湖南省教育厅资助科研项目(21B0390) （21B0390）
湖南省卫生健康委科研计划项目(D202303067601) （D202303067601）
中药粉体与创新药物省部共建国家重点实验室培育基地开放基金项目(21PTKF1016). （21PTKF1016）
