miR-143-3p靶向TREM1基因对狼疮性肾炎小鼠肾小球系膜细胞炎性因子分泌及凋亡的调控机制  

Effects of miR-143-3p on inflammatory factors secretion and apoptosis of mesangial cells in mice with lupus nephritis(LN) through targeting triggering receptor expressed by myeloid cells-1(TREM1)

作　　者：罗慧臣[1] 胡丹慧(指导) 张济[1,3] LUO Hui-Chen;HU Dan-Hui;ZHANG Ji(Department of Rheumatology and Immunology,the First Affiliated Hospital of University of South China,Hengyang 421001,China)

机构地区：[1]南华大学附属第一医院风湿免疫科,衡阳421001 [2]南华大学附属第一医院新生儿科,衡阳421001 [3]南华大学附属第一医院风湿免疫实验室,衡阳421001

出　　处：《中国免疫学杂志》2020年第4期399-404,410,共7页Chinese Journal of Immunology

摘　　要：目的:探讨miR-143-3p靶向髓细胞触发受体1(TREM1)基因对狼疮性肾炎(LN)小鼠肾小球系膜细胞炎性因子分泌及细胞凋亡的影响。方法:免疫组化检测模型及正常小鼠肾组织中TREM1的表达。双荧光素酶报告系统结合Western blot验证miR-143-3p与TREM1的靶向关系。分离纯化模型小鼠肾小球系膜细胞,Western blot检测各组处理后肾小球系膜细胞中NF-κB p65、TNF-α、IL-1β和MCP-1的蛋白表达;CCK8法测定处理后的肾小球系膜细胞活力;流式细胞术检测各组细胞周期及细胞凋亡情况。结果:组织实验显示:与正常小鼠相比,狼疮性肾炎小鼠肾组织中TREM1高表达。同时,miR-143-3p靶向下调TREM1表达。细胞实验显示:与各自的阴性对照相比,sh-TREM1组和miR-143-3p mimic组中NF-κB p65、TNF-α、IL-1β和MCP-1蛋白表达显著下调,系膜细胞活力明显降低,更多细胞停滞在G0/G1期,但凋亡情况无显著变化。结论:miR-143-3p能下调TREM1表达从而抑制LN小鼠肾小球系膜细胞炎性因子生成以及细胞活力的增强,但对其凋亡无显著影响。Objective:To investigate the effects of miR-143-3p on inflammatory factors secretion and apoptosis of mesangial cells in mice with lupus nephritis(LN) through targeting triggering receptor expressed by myeloid cells-1(TREM1).Methods:Immunohistochemistry assay was utilized to detect the expression of TREM1 in kidney tissue of model mice and normal mice.Dual luciferase reporter assay and Western blot were adopted to verify the targeting relationship between miR-143-3p and TREM1.Subsequently mesangial cells of model mice were separated and purified.Western blot was used to determine the protein expression of NF-κB p65,TNF-α,IL-1β and MCP-1 in each group;CCK8 was adopted to measure the viability of mesangial cells;Flow cytometry was used to detect the cell cycle and apoptosis in each group.Results:Tissue experiments indicated that:compared with the normal mice,expression of TREM1 was increased in kidney tissue of mice with LN.miR-143-3p was predicted and confirmed to target TREM1.Cell experiments indicated that:compared with the respective negative control group,protein expression of NF-κB p65,TNF-α,IL-1β and MCP-1 was down-regulated in sh-TREM1 group and miR-143-3p mimic group.At the same time,viability of mesangial cells was decreased,more cells were blocked at G0/G1 phase while no significant difference was observed in apoptosis of mesangial cells compared with the respective negative control group.Conclusion:miR-143-3p could inhibit inflammatory factors and cell viability of mesangial cells through negative-regulating TREM1 in mice with LN,but have no significant impact on apoptosis.

关 键 词：狼疮性肾炎 肾小球系膜细胞 炎性因子 细胞凋亡 

分 类 号：R392.3[医药卫生—免疫学]