Based on the NLRP3/Caspase-1/GSDMD signaling pathway intervene effect of Shenggu Zaizao Pills on pyroptosis of BMSCs
Objective To study the effect of bone reconstruction pills on the level of pyroptosis in BMSCs through NLRP3/Caspase-1/GSDMD signaling pathway.Methods Pyroptosis BMSCs models were prepared,and lentivirus and siRNA technolo-gies were used to transfect pyroptosis BMSCs.The medicated serum of raw bone reconstruction pills was prepared to intervene in the BMSCs of pyroptosis.The above cells were divided into blank group,model group,lentivirus group,siRNA group and drug-containing serum group.CCK-8 was used to detect the activity of BMSCs in each group,and the expression level of GSDMD gene in BMSCs was detected by RT-qPCR.WB was used to detect the expression levels of NLRP3,Caspase-1 and GSDMD in BMSCs.ELISA was used to detect the levels of IL-1β and IL-18 in BMSCs.Results Compared with the blank group,the ex-pression of GSDMD gene in the model group increased(P＜0.01),the protein levels of NLRP3,Caspase-1 and GSDMD were significantly increased(P＜0.05)(P＜0.01)(P＜0.01),and the levels of IL-18 and IL-1 β increased(P＜0.01)(P＜0.01).Compared with the model group,the expression of GSDMD gene in BMSCs in the lentivirus group increased(P＜0.01),the protein levels of NLRP3,Caspase-1 and GSDMD increased(P＜0.05)(P＜0.05)(P＜0.05),the levels of IL-18 and IL-1β increased significantly(P＜0.01)(P＜0.05),and the expression of GSDMD gene in BMSCs in the siRNA group de-creased significantly(P＜0.01),NLRP3,The protein levels of Caspase-1 and GSDMD decreased(P＜0.01)(P＜0.05)(P＜0.05),the levels of IL-18 and IL-1 β decreased significantly(P＜0.01)(P＜0.01),the expression of GSDMD gene in BMSCs in the drug-containing serum group decreased(P＜0.05),NLRP3,Caspase-1,The levels of GSDMD protein de-creased(P＜0.05)(P＜0.05)(P＜0.01),and the levels of IL-1β and IL-18 decreased(P＜0.05)(P＜0.05).Conclu-sion NLRP3 is a key target for regulating pyroptosis of BMSCs,and bone reconstruction pills can regulate the level of pyroptosis of BMSCs by reducing the expression of NLRP3 and inhibiting the NLRP3/Caspase-1/GSDMD signaling pathway.
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