Centre of Organ Transplantation, Xiangya Hospital, Central South University, Changsha, Hunan 410008, P.R. China.
Department of General Surgery, Xiangya Hospital, Central South University, Changsha, Hunan 410008, P.R. China.
Int J Mol Med. 2017 Dec;40(6):1840-1850. doi: 10.3892/ijmm.2017.3194. Epub 2017 Oct 17.
Kidney transplantation is an effective final therapeutic procedure for patients with end-stage kidney failure. Although advanced immunosuppressive therapy is administered following transplantation, certain patients still suffer from acute allograft rejection. MicroRNAs (miRs) have a potential diagnostic and therapeutic value for acute renal allograft rejection; however, their underlying mechanism of action is largely unknown. In the present study, an increased level of miR-650 was identified to be associated with the downregulation of B-cell CLL/lymphoma 11B (BCL11B) expression in acute renal allograft rejection. Furthermore, in vitro study using human renal glomerular endothelial cells (HRGECs) transfected with a miR-650 mimic revealed that key characteristics of acute renal allograft rejection were observed, including apoptosis, the release of cytokines and the chemotaxis of macrophages, while the effects were reduced in HRGECs transfected with a miR-650 inhibitor. The existence of a conserved miR-650 binding site on the 3'-untranslated region of BCL11B mRNA was predicted by computational algorithms and confirmed by a luciferase reporter assay. Knockdown of BCL11B with small interfering RNA (siRNA) significantly increased the apoptotic rate and significantly decreased the proliferation ability of HRGECs compared with the negative control group. HRGECs transfected with a combination of BCL11B siRNA and the miR-650 mimic demonstrated a significant increase in the rate of apoptosis compared with the control. These results suggest that the upregulation of miR-650 contributes to the development of acute renal allograft rejection by suppression of BCL11B, which leads to apoptosis and inflammatory responses. Thus, miR-650 and BCL11B may represent potential therapeutic targets for the prevention of acute renal allograft rejection.
肾移植是治疗终末期肾衰竭患者的有效终末治疗手段。尽管移植后会进行先进的免疫抑制治疗,但某些患者仍会发生急性同种异体移植排斥反应。微小 RNA(miRs)在急性肾同种异体移植排斥反应的诊断和治疗中具有潜在价值;然而,其作用机制在很大程度上尚不清楚。本研究发现,miR-650 水平升高与急性肾同种异体移植排斥反应中 B 细胞 CLL/淋巴瘤 11B(BCL11B)表达下调有关。此外,在转染 miR-650 模拟物的人肾肾小球内皮细胞(HRGECs)的体外研究中,观察到急性肾同种异体移植排斥的关键特征,包括细胞凋亡、细胞因子释放和巨噬细胞趋化作用,而转染 miR-650 抑制剂的 HRGECs 中这些作用则减弱。计算算法预测 BCL11B mRNA 3'-非翻译区存在保守的 miR-650 结合位点,并通过荧光素酶报告基因检测得到证实。与阴性对照组相比,小干扰 RNA(siRNA)敲低 BCL11B 显著增加 HRGECs 的凋亡率,显著降低其增殖能力。与对照组相比,转染 BCL11B siRNA 和 miR-650 模拟物的 HRGECs 凋亡率显著增加。这些结果表明,miR-650 通过抑制 BCL11B 促进急性肾同种异体移植排斥反应的发生,从而导致细胞凋亡和炎症反应。因此,miR-650 和 BCL11B 可能成为预防急性肾同种异体移植排斥反应的潜在治疗靶点。