Mechanism of Serum Containing Anwei Decoction(安胃汤)Regulating Pyroptosis of NLRP3-Caspase-1-IL-1βSignal Axis of MC Cells
Objective To investigate the effects of Anwei Decoction(安胃汤)on NACHT-LRR-PYD-containing proteins 3 inflammasome(NLRP3)-cysteine-requiring aspartate protease-1(Caspase-1)-interleukin-1β(IL-1 β)signaling pathway related factors and pyroptosis in mast cells(MC)cells,and reveal the anti-chronic atrophic gastritis(CAG)mechanism of Anwei Decoction.Methods MC cells were divided into 4 groups:blank group,Anwei Decoction group,Anwei Decoction+Z-YVVD-FMK group and Z-YVVD-FMK group.CCK8 was used to detect cell proliferation at 12 h,24 h and 48 h and to de-termine the optimal detection time point and the proportion of drug-containing serum.The lactate dehydrogenase(LDH)activity in cell supernatant of each group was detected.AO-EB staining was used to detect the integrity of cell membrane and nuclear morphology.The levels of serum interleukin-18(IL-18)and IL-1 β were detected by ELISA.Real-time PCR was used to detect the expressions of NLRP3,Caspase-1,gasdermin D(GSDMD),IL-18 and IL-1 β genes.Western-blot was used to detect the expressions of NLRP3,Caspase-1 and GSDMD proteins.Results The 48 h time point and 10％drug-containing ser-um ratio were chosen to the follow-up experiment.Compared with that of the other three groups,the activity of LDH in the An-wei Decoction group was increased(P＜0.01).AO-EB staining in the administration groups showed swelling and deformation of the cell membrane and changes in nuclear morphology.The results of ELISA showed that compared with those of the blank group,the levels of IL-18 and IL-1 β in the Anwei Decoction group were increased(P＜0.01),and the levels of IL-18 and IL-1 β in the Z-YVVD-FMK group were decreased(P＜0.01),and the difference was statistically significant.Real-time PCR showed that compared with those of the blank group,the expression levels of NLRP3mRNA,Caspase-1mRNA,GSDMDmR-NA,IL-18mRNA and IL-1 βmRNA in the Anwei Decoction group were increased(P＜0.01,P＜0.05).The expression levels of GSDMDmRNA,IL-18mRNA and IL-1 βmRNA in the blocker group were decreased(P＜0.01),and the difference was sta-tistically significant.The results of Western-blot showed that compared with those of the blank group,the expressions of NLRP3/Actin and GSDMD/Actin in the Anwei Decoction group decreased(P＜0.01),and the expression of Caspase-1/Actin in the Z-YVVD-FMK group decreased(P＜0.01).Conclusion The anti-CAG effect of Anwei Decoction may be related to the re-versal of the pathological state of gastric mucosal cells induced by pyroptosis by regulating NLRP3-Caspase-1-IL-1 β signal transduction pathway.
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