南方医科大学学报 ›› 2016, Vol. 36 ›› Issue (03): 419-.

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miR-663在肾移植急性排斥反应中的调控作用

刘小友,张婕,梁洁,刘永光,胡建敏,姜正尧,郭泽锋   

  • 出版日期:2016-03-20 发布日期:2016-03-20

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

  • Online:2016-03-20 Published:2016-03-20

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。

Abstract: 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-663levels in renal transplant recipients with and without AR. MTT assay and Annexin V-FITC assay were employed to examinethe viability and apoptosis of human renal glomerular endothelial cells (HRGEC) treated with a miR-663 mimic or a miR-663inhibitor, 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 thechemotactic capability of macrophages. Results Serum miR-663 level was significantly higher in renal transplant recipientswith AR than in those without AR. The miR-663 mimic significantly inhibited the viability of HRGECs and increase the cellapoptosis rate, while miR-663 inhibitor suppressed the cell apoptosis. The miR-663 mimic increased the expression levels ofinflammation-related cytokines and enhanced the chemotactic capability of macrophages. Conclusion miR-663 might playimportant roles in acute renal graft rejection and may become a therapeutic target for treating AR.