摘要
目的 探讨miR-22-3p对同型半胱氨酸(Hey)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(0 μmol/L Hey)与 Hey 组(100 μmol/L Hey),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 Hey 诱导下 VSMC 的变化.结果 与 Control 组相比,Hey 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Hey诱导的VSMC焦亡.
Abstract
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(0 μmol/L Hey)and a Hey group(100 μmol/L Hey).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)-1 β and 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-1 β and 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-1β and 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-1β and IL-18(P＜0.05).Conclusions MiR-22-3p may delay Hcy-induced VSMC pyroptosis.
基金项目
国家自然科学基金项目(U21A20343)
国家自然科学基金项目(82370293)
国家自然科学基金项目(82270492)
宁夏回族自治区重点研发计划重点项目(2023BEG02074)
宁夏回族自治区重点研发计划重点项目(2022BFH02013)
宁夏回族自治区重点研发计划重点项目(2021BEG02033)
宁夏回族自治区重点研发计划重点项目(2020BFH02001)
维普
万方数据