消皮素B在人主动脉夹层组织中的表达变化及对主动脉平滑肌细胞焦亡、炎症反应的影响  

Expression of gasdermin B in human aortic dissection tissues and its effect on pyroptosis and inflammatory response of human aortic smooth muscle cells

作　　者：沈小艳 谢孝平 李博文[1] 王志维[1] SHEN Xiaoyan;XIE Xiaoping;LI Bowen;WANG Zhiwei(Department of Cardiothoracic Surgery,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院心血管外科,武汉430060

出　　处：《山东医药》2024年第11期6-10,共5页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(82070481,82200522)。

摘　　要：目的观察人主动脉夹层组织消皮素B(GSDMB)表达变化,并探讨GSDMB对人主动脉平滑肌细胞(HASMCs)焦亡和炎症反应的影响。方法收集主动脉夹层患者的主动脉夹层组织、器官捐献者的主动脉组织(正常对照组织)各8例份,采用Western blotting法检测GSDMB蛋白表达。将对数生长期的HASMCs分为NC-siRNA组(转染NC-siRNA)、脂多糖(LPS)组(LPS作用)、GSDMB-siRNA组(转染GSDMB-siRNA)、GSDMB-siRNA+LPS组(LPS作用后转染GSDMB-siRNA),另将HASMCs分为空载质粒组(转染空载质粒)、LPS作用组(LPS作用)、GSDMB过表达组(转染GSDMB过表达质粒)、GSDMB过表达+LPS组(LPS作用后转染GSDMB过表达质粒),采用Western blotting法检测细胞GSDMB、半胱氨酸蛋白酶4(Caspase-4)、消皮素D(GSDMD)、N-消皮素D(N-GSDMD)、基质金属蛋白酶9(MMP-9)、基质金属蛋白酶2(MMP-2)蛋白表达,流式细胞术检测细胞焦亡率,ELISA法检测细胞上清液白细胞介素18(IL-18)、白细胞介素1β(IL-1β)水平。结果主动脉夹层组织与正常对照组织GSDMB蛋白相对表达量分别为0.88±0.16、0.62±0.15,二者比较P<0.05。与NC-siRNA组比较,LPS组GSDMB、Caspase-4、GSDMD、N-GSDMD、MMP-9、MMP-2蛋白相对表达量和细胞焦亡率及细胞上清液IL-18、IL-1β水平均升高,GSDMB-siRNA组仅GSDMB蛋白相对表达量降低(P均<0.05)。与LPS组比较,GSDMB-siRNA+LPS组GSDMB、Caspase-4、GSDMD、N-GSDMD、MMP-9、MMP-2蛋白相对表达量和细胞焦亡率及细胞上清液IL-18、IL-1β水平均降低(P均<0.05)。与空载质粒组比较,LPS作用组GSDMB、Caspase-4、GSDMD、N-GSDMD、MMP-9、MMP-2蛋白相对表达量和细胞焦亡率及细胞上清液IL-18、IL-1β水平均升高(P均<0.05),GSDMB过表达组仅GSDMB蛋白相对表达量升高(P<0.05)。与LPS作用组比较,GSDMB过表达+LPS组GSDMB、Caspase-4、GSDMD、N-GSDMD、MMP-9、MMP-2蛋白相对表达量和细胞焦亡率及细胞上清液IL-18、IL-1β水平均升高(P均<0.05)。结论人主动脉夹层组织GSDMB表�Objective To observe the expression changes of gasdermin B(GSDMB)in human aortic dissection tissues 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 NCsiRNA 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-1βin the cell supernatant 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

关 键 词：消皮素B 主动脉夹层 人主动脉平滑肌细胞 细胞焦亡 炎症反应 半胱氨酸蛋白酶4 

分 类 号：R543.1[医药卫生—心血管疾病]