基于α7nAchR和Caspase-11/GSDMD通路探讨芒柄花素对ox-LDL诱导的AS泡沫细胞焦亡的调控机制OA北大核心CSTPCD
Regulation Mechanism of Formononetin on ox-LDL-Induced AS Foam Cells Pyroptosis Based on α7nAchR and Caspase-11/GSDMD Pathway
目的:探讨芒柄花素对氧化低密度脂蛋白(ox-LDL)诱导动脉粥样硬化(AS)泡沫细胞焦亡的抑制作用及其潜在机制.方法:通过ox-LDL诱导RAW264.7 细胞构建AS泡沫细胞模型,采用油红O染色验证细胞模型建立成功,实验设正常对照组、模型组、芒柄花素组、芒柄花素+α7nAchR拮抗剂组、芒柄花素+α7nAchR激动剂组.油红O染色检测细胞脂肪蓄积,CCK-8 检测细胞的增殖活力,Transwell检测细胞的迁移能力,RT-qPCR、免疫荧光和Western Blot检测焦亡相关蛋白Caspase-11、GSDMD-N、GSDMD表达.沉默α7nAchR后,Western Blot检测芒柄花素对上述焦亡相关蛋白的影响.结果:与正常对照组比较,模型组细胞分化严重且充满大量红色脂质颗粒,细胞增殖活力、迁移能力以及Caspase-11、GSDMD-N、GSDMD mRNA和蛋白表达显著升高(P<0.05).与模型组比较,芒柄花素组细胞脂肪蓄积改善,增殖活力、迁移能力以及Caspase-11、GSDMD-N、GSDMD mRNA和蛋白表达显著降低(P<0.05).α7nAchR拮抗剂可逆转芒柄花素的作用,α7nAchR激动剂可增强芒柄花素的作用(P<0.05).α7nAchR沉默后,Caspase-11、GSDMD-N、GSDMD的表达显著高于模型组,而加入芒柄花素处理后显著逆转上述现象.结论:芒柄花素可有效减少ox-LDL诱导的巨噬细胞损伤,其机制可能与促进α7nAchR的表达和抑制Caspase-11/GSDMD通路介导的巨噬细胞焦亡有关.
Objective:To investigate the inhibitory effect and its potential mechanism of formononetin on oxidized low-density lipo-protein(ox-LDL)-induced pyroptosis of atherosclerotic(AS)foam cells.Methods:RAW264.7 cells were induced by ox-LDL to establish AS foam cell model.The successful establishment of foam cell model was verified by oil red O staining.The cells were divided into nor-mal control group,model group,formononetin group,formononetin with α7nAchR antagonist group,formononetin with α7nAchR agonist group.The fat accumulation of cells was detected by oil red O staining,CCK-8 was used to detect the proliferation activity of cells,Tran-swell was used to detect the migration ability of cells,RT-qPCR,immunofluorescence and Western Blot were used to detect the expres-sions of pyroptosia-related proteins Caspase-11,GSDMD-N,GSDMD.After silencing α7nAchR,Western Blot was used to detect the effect of formononetin on the above-mentioned pyroptosis related proteins.Results:Compared with the normal control group,the cells in the model group were severely differentiated and filled with a large number of red lipid particles.The cell proliferation activity,migration ability,as well as the expressions of Caspase-11,GSDMD-N,GSDMD mRNA and proteins were significantly increased(P<0.05).Com-pared with the model group,the fat accumulation of cells in formononetin group was improved,and the proliferation activity,migration a-bility,as well as the mRNA and protein expressions of Caspase-11,GSDMD-N,GSDMD were significantly decreased(P<0.05).The α7nAchR antagonist could reverse the effect of formononetin,while the α7nAchR agonist could enhance the effect of formononetin(P<0.05).After silencing α7nAchR,the expressions of Caspase-11,GSDMD-N and GSDMD were significantly higher than those in the model group,while the addition of formononetin treatment significantly reversed the above phenomena.Conclusion:Formononetin can ef-fectively reduce the macrophage injury induced by ox-LDL.The mechanism may be related to promoting the expression of α7nAchR and inhibiting the pyroptosis of macrophages mediated by the Caspase-11/GSDMD pathway.
吴金燕;李世容;张敏;唐梅;王文秀;曹丽平;何前松
贵州中医药大学第一临床医学院,贵州 贵阳 550001贵州省人民医院神经内科,贵州 贵阳 550001贵州中医药大学第一附属医院神经内科,贵州 贵阳 550001贵州中医药大学第一附属医院神经内科,贵州 贵阳 550001贵州中医药大学第一附属医院神经内科,贵州 贵阳 550001贵州中医药大学第一附属医院神经内科,贵州 贵阳 550001贵州中医药大学第一临床医学院,贵州 贵阳 550001||贵州中医药大学第一附属医院神经内科,贵州 贵阳 550001
医药卫生
芒柄花素细胞焦亡Caspase-11/GSDMD信号通路ox-LDL
FormonectinPyroptosisCaspase-11/GSDMD Signal pathwayox-LDL
《中药材》 2025 (12)
3135-3141,7
国家自然科学基金项目(82160850)贵州省科技计划项目(黔科合基础-ZK[2021]一般541)
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