miR-34a对高糖状态下小鼠巨噬细胞系RAW264.7极化及炎性因子分泌的影响  

Effects of miR-34a on polarization and inflammatory factor content of mouse macrophage cell line RAW264.7 under high glucose conditions

作　　者：张凤云 赵志浩 张卓琦 ZHANG Fengyun;ZHAO Zhihao;ZHANG Zhuoqi(Department of Cardiology,Affiliated Hospital of Xuzhou Medical University,Xuzhou 221000,China)

机构地区：[1]徐州医科大学附属医院心内科,江苏徐州221000

出　　处：《基础医学与临床》2023年第12期1808-1813,共6页Basic and Clinical Medicine

基　　金：徐州市推动科技创新项目(KC19062)。

摘　　要：目的探讨微小RNA-34a(miR-34a)对高糖条件下小鼠巨噬细胞系极化和炎性因子表达的影响。方法将小鼠巨噬细胞在高糖条件下(25 mmol/L)下培养3至28 d,RT-qPCR检测高糖培养时巨噬细胞中miR-34a表达及M1巨噬细胞标志物(iNOS和MCP-1)和M2巨噬细胞标志物(Arg-1)的基因表达。转染miR-34a模拟物或抑制剂后,ELISA检测炎性相关因子(IL-6、IL-1β和TNF-α)的分泌。Western blot检测miR-34a靶蛋白1型跨膜糖蛋白Notch1的表达。结果在高糖条件下,巨噬细胞中miR-34a以及M1型巨噬细胞标志物(iNOS和MCP-1)基因表达量逐渐增长。过表达miR-34a后,促炎因子IL-6、IL-1β和TNF-α分泌增加,iNOS和MCP-1基因表达量也明显增加,而沉默miR-34a的表达后,炎性因子(IL-6、IL-1β和TNF-α)分泌及M1型巨噬细胞标记物(iNOS和MCP-1)的表达量降低(P<0.05)。沉默Notch1表达后,miR-34a及IL-6、IL-1β和TNF-α分泌及iNOS和MCP-1表达量下降。结论慢性高糖刺激miR-34a表达,后者促进巨噬细胞向M1型极化和促炎因子的分泌。Objective To detect the role of miR-34a on macrophage inflammation and polarization under high glucose conditions.Methods Mouse macrophages were collected and cultured during high glucose for 3-28 days.RT-qPCR was used to detect the expression of miR-34a,iNOS,MCP-1 and Arg-1 mRNA.Then miR-34a was over-expressed or silenced,ELISA was used to detect the expression of IL-6,IL-1β,TNF-αand qRT-PCR was used to detect the expression of iNOS and MCP-1 mRNA.Western blot was used to detect the expression of Notch1.Results Expression of miR-34a increased under high glucose conditions in RAW264.7 cells continuously.Over-expression of miR-34a promoted the expression of MCP-1 and iNOS observed by RT-qPCR and increased the expression of IL-6,IL-1βand TNF-αdetected by ELISA.Further studies showed that siRNA-Notch1 down-regulated the expression of miR-34a,MCP-1,iNOS,IL-6,IL-1βand TNF-α.Conclusions Chronic high glucose condition stimulates the expression of miR-34a which promotes M1 macrophage polarization and releasing of pro-inflammatory factors.

关 键 词：MIR-34A 巨噬细胞 高糖 极化 炎性反应 

分 类 号：R587.1[医药卫生—内分泌]