miR-22-3p靶向GSDMD抑制同型半胱氨酸诱导的平滑肌细胞焦亡  

MiR-22-3p targets gasdermin D to inhibit homocysteine-induced pyroptosis of vascular smooth muscle cells

作　　者：钟颖怡 丁宁[2] 王浥尘 刘超 董治锋 马胜超 熊建团 郝银菊 白志刚[4] 姜怡邓 ZHONG Yingyi;DING Ning;WANG Yichen;LIU Chao;DONG Zhifeng;MA Shengchao;XIONG Jiantuan;HAO Yinju;BAI Zhigang;JIANG Yideng(National Health Commission Key Laboratory of Metabolic Cardiovascular Disease Research,Yinchuan 750004,China;School of Basic Medicine,Ningxia Medical University,Yinchuan 750004;School of Clinical Medicine,Ningxia Medical University,Yinchuan 750004;Ningxia Hui Autonomous Region People’s Hospital,Yinchuan 750004)

机构地区：[1]国家卫生健康委代谢性心血管疾病研究重点实验室,银川750004 [2]宁夏医科大学基础医学院,银川750004 [3]宁夏医科大学临床医学院,银川750004 [4]宁夏回族自治区人民医院,银川750004

出　　处：《中国比较医学杂志》2024年第9期12-18,共7页Chinese Journal of Comparative Medicine

基　　金：国家自然科学基金项目(U21A20343,82370293,82270492);宁夏回族自治区重点研发计划重点项目(2023BEG02074,2022BFH02013,2021BEG02033,2020BFH02001)。

摘　　要：目的探讨miR-22-3p对同型半胱氨酸(Hcy)诱导血管平滑肌细胞焦亡的影响。方法体外培养人血管平滑肌细胞(VSMC),分为Control组(0μmol/L Hcy)与Hcy组(100μmol/L Hcy),Western blot检测细胞中pro Caspase-1、GSDMD、N-GSDMD、NLRP3表达水平,qRT-PCR检测miR-22-3p表达情况;ELISA检测上清中IL-1β、IL-18的浓度。转染miR-22-3p对照(miR-22-3p-NC)、miR-22-3p的模拟物(miR-22-3p-mimic)及抑制物(miR-22-3p-inhibitor)后观察Hcy诱导下VSMC的变化。结果与Control组相比,Hcy组VSMC中pro Caspase-1、GSDMD、N-GSDMD、NLRP3表达水平增加(P<0.05),IL-1β、IL-18浓度更高(P<0.01),miR-22-3p相对表达水平低(P<0.01);转染miR-22-3p-mimic后,VSMC中pro Caspase-1、GSDMD、N-GSDMD、NLRP3表达水平显著降低(P<0.01),IL-1β、IL-18浓度降低(P<0.01);转染miR-22-3p-inhibitor后VSMC中pro Caspase-1、GSDMD、N-GSDMD、NLRP3表达水平显著增加(P<0.01),IL-1β、IL-18浓度升高(P<0.05)。结论miR-22-3p可抑制Hcy诱导的VSMC焦亡。Objective To investigate the effect of miR-22-3p on pyroptosis of vascular smooth muscle cells(VSMCs)induced by homocysteine(Hcy).Methods Human VSMCs were cultured in vitro and divided into a Control group(0μmol/L Hcy)and a Hcy group(100μmol/L Hcy).After intervention,expression levels of pro Caspase-1,gasdermin D(GSDMD),N-GSDMD,and NLR family pyrin domain containing 3(NLRP3)were detected by Western blot.MiR-22-3p expression was determined by quantitative real-time reverse-transcription polymerase chain reaction.Interleukin(IL)-1βand IL-18 levels in the supernatant were measured by enzyme-linked immunosorbent assay.Cells were also transfected with control miR-22-3p(miR-22-3p-NC),miR-22-3p-mimic,and miR-22-3p-inhibitor,to observe the effects on VSMC pyroptosis induced by Hcy.Results Expression levels of pro Caspase-1,GSDMD,N-GSDMD,and NLRP3 in VSMCs were increased(P<0.05),IL-1βand IL-18 levels were increased(P<0.01),and the relative expression level of miR-22-3p was reduced(P<0.01)in the Hcy group compared with the Control group.Transfection with miR-22-3p-mimic significantly decreased the expression levels of pro Caspase-1,GSDMD,N-GSDMD,and NLRP3 in VSMCs(P<0.01),and significantly decreased levels of IL-1βand IL-18(P<0.01),while transfection with miR-22-3p-inhibitor significantly increased the expression levels of pro Caspase-1,GSDMD,N-GSDMD,and NLRP3 in VSMCs(P<0.01)and significantly increased the levels of IL-1βand IL-18(P<0.05).Conclusions MiR-22-3p may delay Hcy-induced VSMC pyroptosis.

关 键 词：动脉粥样硬化 血管平滑肌细胞 焦亡 miR-22-3p 同型半胱氨酸 

分 类 号：R-33[医药卫生]