硫化氢通过下调NLRP3/caspase-1信号通路抑制氧化型低密度脂蛋白诱导的血管内皮细胞焦亡  

Hydrogen sulfide inhibits oxidized low-density lipoprotein-induced pyroptosis in vascular endothelial cells by down-regulating NLRP3/caspase-1 signaling pathway

作　　者：何婷婷 张旭琳 贾珍丽 王昀 马克涛[1,2] 李玲 张亮[1,2] HE Ting-ting;ZHANG Xu-lin;JIA Zhen-li;WANG Yun;MA Ke-tao;LI Ling;ZHANG Liang(Department of Basic Medicine,School of Medicine,Shihezi University,Shihezi 832000,China;NHC Key Laboratory of Prevention and Treatment of Central Asia High Incidence Diseases,Shihezi 832000,China;Medical Teaching Experimental Center,School of Medicine,Shihezi University,Shihezi 832000,China)

机构地区：[1]石河子大学医学院基础医学系,新疆石河子832000 [2]国家卫生健康委中亚高发病防治重点实验室,新疆石河子832000 [3]石河子大学医学院医学教学实验中心,新疆石河子832000

出　　处：《中国病理生理杂志》2021年第10期1738-1746,共9页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81600325);中国医学科学院中央级公益性科研院所基本科研业务费专项资金资助项目(No.2020-PT330-003);国家卫生健康委中亚高发病防治重点实验室开放课题(No.KF202107);石河子大学青年创新人才计划项目(No.CXBJ201906)。

摘　　要：目的:探讨缓释型硫化氢(H2S)供体GYY4137对氧化型低密度脂蛋白(oxLDL)诱导的血管内皮细胞焦亡的抑制作用及其分子机制。方法:以人脐静脉内皮细胞(HUVECs)为研究对象,将其分为空白对照组、GYY4137干预组、oxLDL刺激组和oxLDL+GYY4137干预组。各组HUVECs经不同药物干预指定时间后,采用CCK-8法检测各组细胞活力;使用乳酸脱氢酶(LDH)释放测定和Hoechst 33342/碘化丙啶(PI)双荧光标记检测各组细胞死亡情况;采用亚甲基蓝比色法检测各组细胞中的H2S含量;采用Western blot实验观察各组细胞中细胞焦亡信号通路标志蛋白的表达。结果:oxLDL(50和100 mg/L)可使HUVECs活力显著降低(P<0.05或P<0.01),LDH释放及PI阳性细胞比例显著增加(P<0.05或P<0.01),且细胞中NLRP3、caspase-1(p20亚单位)、gasdermin D(GSDMD)、GSDMD-N及白细胞介素18(IL-18)蛋白水平显著升高(P<0.05或P<0.01),即HUVECs发生焦亡。而GYY4137干预则可降低HUVECs中NLRP3、caspase-1(p20)、GSDMD、GSDMD-N及IL-18蛋白水平(P<0.05),抑制oxLDL诱导的细胞焦亡(P<0.05),并使细胞活力增强(P<0.05)。结论:H2S供体GYY4137可通过下调NLRP3/caspase-1细胞焦亡信号通路而抑制oxLDL诱导的血管内皮细胞焦亡。AIM:To investigate the inhibitory effect of slow-releasing hydrogen sulfide(H2 S)donor GYY4137 on oxidized low-density lipoprotein(oxLDL)-induced pyroptosis of vascular endothelial cells and its potential mechanisms.METHODS:Human umbilical vein endothelial cells(HUVECs)were incubated with oxLDL or/and GYY4137 at different concentrations or indicated concentration for indicated time periods according to different experimental requirements. The HUVECs were randomly divided into the following 4 groups:(1)normal control group:HUVECs treated with endothelial cell medium containing 1% fetal bovine serum were used as control;(2)GYY4137 treatment group(GYY4137 group):HUVECs were treated with GYY4137 at indicated concentration for indicated time periods;(3)oxLDL treatment group(oxLDL group):HUVECs were treated with oxLDL at different concentrations for indicated time periods;(4)oxLDL+GYY4137 treatment group(oxLDL+GYY4137 group):HUVECs were pretreated with GYY4137(200 μmol/L)for 4 h at desired concentration,and then incubated with oxLDL(100 mg/L)for indicated time periods in the presence of GYY4137. After the indicated treatments,the effects of oxLDL and GYY4137 on the viability of HUVECs were measured by CCK-8 assay. Pyroptotic cell death in each group was evaluated by lactate dehydrogenase(LDH)release and Hoechst33342/propidium iodide(PI)staining. The H2 S production in each group was measured by methylene blue method and spectrophotometry. Western blot was performed to detect the expression of specific markers of pyroptosis in each group.RESULTS:Treatment with oxLDL(50 and 100 mg/L)significantly decreased the viability of HUVECs(P<0. 05 or P<0. 01),increased LDH release and percentage of PI positive cells(P<0. 05 or P<0. 01),and up-regulated the protein levels of NLRP3,caspase-1(p20),gasdermin D(GSDMD),GSDMD-N and interleukin-18(IL-18)in the HUVECs(P<0. 05 or P<0. 01),indicating the pyroptosis of HUVECs. However,GYY4137 pre-treatment inhibited the pyroptosis of oxLDL-incubated HUVECs as evidenced by decreasing LDH rele

关 键 词：硫化氢 动脉粥样硬化 血管内皮细胞 细胞焦亡 NLRP3/caspase-1信号通路 

分 类 号：R543.5[医药卫生—心血管疾病] R363.2[医药卫生—内科学]