Mechanism of Juanbi Chushi Decoction Regulating the Escape of Mitochondrial DNA to Prevent Macrophage Pyroptosis
Objective To investigate the effect and mechanism of Juanbi Chushi decoction on macrophage pyroptosis via regulating the escape of cytoplasmic mitochondrial DNA(mtDNA).Methods Human mononuclear leukemia cells were induced to differentiate into macrophages.Macrophages treated by mtDNA were divided into 3groups:control group,LPS+ATP group,and LPS+ATP+mtDNA group.Macrophages treated by Juanbi Chushi decoction were divided into 4groups:control group,Juanbi Chushi decoction group,LPS+ATP group,and LPS+ATP+Juanbi Chushi decoction group.Cytoplasmic mtDNA was extracted and its copy number was detected by re-al-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Cell viability was detected by CCK8.The levels of interleu-kin-1 β(IL-1 β)and tumor necrosis factor-α(TNF-α)in supernatant were detected by enzyme-linked immunosorbent assay(ELISA).Lactate dehydrogenase(LDH)level was detected by LDH cytotoxicity assay kit.Annexin+/7 A AD+cells were detected by flow cytometry.The expression levels of NLRP3,activated caspase-1 and GSDMD-N protein were detected by Western blot.Results MtDNA could promote macrophage pyroptosis.Compared with control group,the cytoplasmic mtDNA level and macrophage pyroptosis were significantly increased in LPS+ATP group.Compared with LPS+ATP group,Juanbi Chushi decoction decreased cytoplasmic mtD-NA level and macrophage pyroptosis.The expression levels of NLRP3,activated caspase-1 and GSDMD-N protein were increased in pyroptotic macrophages,while these protein expression levels were significantly decreased after the treatment of Juanbi Chushi decoction.Conclusion Juanbi Chushi decoction may reduce macrophage pyroptosis through NLRP3/caspase-1/GSDMD signaling pathway by re-ducing cytoplasmic mtDNA level.
万方数据
维普
