Expression of gasdermin B in human aortic dissection tissues and its effect on pyroptosis and inflammatory response of human aortic smooth muscle cells
Objective To observe the expression changes of gasdermin B(GSDMB)in human aortic dissection tis-sues and its effect on pyroptosis and inflammatory response of human aortic smooth muscle cells(HASMCs).Methods Eight cases each of the aortic tissues from patients with aortic dissection and from organ donors were collected and GSDMB protein expression was detected by Western blotting.HASMCs in the logarithmic growth phase were divided into the NC-siRNA group(transfected with NC-siRNA),lipopolysaccharide(LPS)group(LPS stimulation),GSDMB-siRNA group(transfected with GSDMB-siRNA),GSDMB-siRNA+LPS group(transfected with GSDMB-siRNA after LPS stimulation),or HASMCs were divided into the empty plasmid group(transfected with empty plasmid),LPS-stimulated group(LPS stimulation),GSDMB overexpression plasmid group(transfected with GSDMB overexpression plasmid),and GSDMB overexpression plasmid+LPS-stimulated group(transfected with GSDMB overexpression plasmid after LPS stimulation),respectively.Western blotting was used to detect the protein expression of GSDMB,Caspase-4,Gasdermin D(GSDMD),N-gasdermin D(N-GSDMD),matrix metalloproteinase-9(MMP-9),and matrix metalloproteinase-2(MMP-2),and flow cytometry was used to detect the pyroptosis rate in HASMCs.ELISA was performed to detect the levels of interleukin(IL)IL-18 and interleukin-1β(IL-1β)in the cell supernatant.Results The relative expression levels of GSDMB protein in the aortic dissection tissues and normal control tissues were 0.88±0.16 and 0.62±0.15,respectively,with statistically significant difference(P<0.05).Compared with the NC-siRNA group,the relative expression levels of GSDMB,Caspase-4,GSDMD,N-GSDMD,MMP-9,MMP-2 proteins,the pyroptosis rate,and the levels of IL-18 and IL-1β in the cell su-pernatant increased in the LPS group(all P<0.05),and only the relative expression of GSDMB protein was reduced in the GSDMB-siRNA group(P<0.05).Compared with the LPS group,the relative expression levels of GSDMB,Caspase-4,GSDMD,N-GSDMD,MMP-9,and MMP-2 proteins,the pyroptosis rate,and the levels of IL-18 and IL-1β in the cell su-pernatant of the GSDMB-siRNA+LPS group all decreased(all P<0.05).Compared with the empty plasmid group,the rel-ative expression levels of GSDMB,Caspase-4,GSDMD,N-GSDMD,MMP-9,MMP-2 proteins,the pyroptosis rate,and cell supernatant IL-18 and IL-1β levels all increased in the LPS-stimulated group(all P<0.05),and only the relative ex-pression of GSDMB protein increased in the GSDMB overexpression plasmid group(P<0.05).Compared with the LPS-stimulated group,the relative expression levels of GSDMB,Caspase-4,GSDMD,N-GSDMD,MMP-9,MMP-2 proteins,the pyroptosis rate,and the levels of IL-18 and IL-1β in the cellular supernatant of the GSDMB overexpression plasmid+ LPS group all increased(all P<0.05).Conclusion GSDMB expression increased in human aortic dissection tissues,and GSDMB did not affect the pyroptosis and inflammatory response of normal HASMCs but positively regulated LPS-in-duced cellular pyroptosis and inflammatory response of HASMCs via a Caspase-4-mediated non-classical pathway.
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