miR-663在肾移植急性排斥反应中的调控作用  

Role of miR-663 in acute renal graft rejection: an in vitro study

作　　者：刘小友[1] 张婕[1] 梁洁[2] 刘永光[1] 胡建敏[1] 姜正尧 郭泽锋[1] 

机构地区：[1]南方医科大学珠江医院器官移植中心,广东广州510282 [2]南方医科大学南方医院门诊部,广东广州510515

出　　处：《南方医科大学学报》2016年第3期419-422,436,共5页Journal of Southern Medical University

基　　金：广东省教育厅科技创新项目(2013KJCX0044)

摘　　要：目的比较肾移植急性排斥(acute rejection,AR)病人及非AR病人血清miR-663表达水平,在细胞水平上探讨miR-663参与肾移植AR的调控作用,为临床早期诊治AR提供新思路。方法 Real time PCR检测肾移植AR病人及非AR病人血清miR-663表达水平。设置miR-663 mimic组、miR-663 inhibitor组、阴性对照组及空白对照组;MTT及Annexin V-FITC分别检测过表达miR-663和抑制miR-663表达对人肾小球内皮细胞(HRGEC)生存率和凋亡率的影响;ELISA检测过表达miR-663和抑制miR-663表达对IL-6、IFN-γ、CCL-2及TNF-α表达水平的影响;Transwell实验检测过表达miR-663和抑制miR-663表达对巨噬细胞趋化性的影响。结果 AR组患者血清miR-663表达水平明显较非AR组的肾移植患者升高(4.73±0.28 vs 1.06±0.04;P<0.01)。MTT显示过表达miR-663可以降低HRGEC的生存率并且明显增加其凋亡率,而抑制miR-663则可以降低HRGEC凋亡。过表达miR-663可以明显提高相关炎症因子的表达,同时明显增加巨噬细胞的趋化性。结论 miR-663在肾移植AR过程中发挥着重要作用,可做为早期诊断AR的外周血标志物,并有望成为治疗肾移植AR的一个潜在分子靶点。Objective To compare the serum miR- 663 levels in renal transplant patients with and without acute rejection（AR）and explore the role of miR- 663 acute renal graft rejection. Methods Real time- PCR was used to determine serum miR- 663 levels in renal transplant recipients with and without AR. MTT assay and Annexin V- FITC assay were employed to examine the viability and apoptosis of human renal glomerular endothelial cells（HRGEC） treated with a miR-663 mimic or a miR-663 inhibitor, and ELISA was performed to detect the expression of inflammation-related cytokines including IL-6, IFN-γ, CCL-2and TNF- α in the cells. Transwell assay was used to examine the effect of miR- 663 mimic and miR- 663 inhibitor on the chemotactic capability of macrophages. Results Serum miR- 663 level was significantly higher in renal transplant recipients with AR than in those without AR. The miR- 663 mimic significantly inhibited the viability of HRGECs and increase the cell apoptosis rate, while miR- 663 inhibitor suppressed the cell apoptosis. The miR- 663 mimic increased the expression levels of inflammation- related cytokines and enhanced the chemotactic capability of macrophages. Conclusion miR- 663 might play important roles in acute renal graft rejection and may become a therapeutic target for treating AR.

关 键 词：肾移植 急性排斥反应 miR-663 

分 类 号：R699.2[医药卫生—泌尿科学]