NK细胞源miR-21在移植肾小管损伤中的作用及其核内调控机制研究

移植肾延迟复功（DGF）是公民死亡后供体来源（DCD）肾移植术后早期最常见的并发症，缺血再灌注（I/R）损伤是发生DGF的主要原因，肾小管损伤是其中心环节。相关研究和本项目组前期工作显示NK细胞参与DGF病理进程；NK细胞与缺糖缺氧HK2细胞共培养后外泌体分泌增加，且所含miR-21显著增多，同时共培养HK2胞核内miR-21信号增强，伴随自噬减少和凋亡增加。已知microRNA具备入核调控基因转录、修饰功能，而miR-21在细胞核内调控作用仍无文献报道。因此，研究组将通过离体共培养和在体小鼠肾移植I/R模型，结合基因编辑、RNA示踪、RIP-Mass/Seq等手段，对NK源miR-21在肾小管上皮细胞损伤中的量效关系、核内通路进行鉴定和功能分析，确定核内调控机制和靶点，并通过干预拮抗性试验进行验证，评估NK源miR-21在减少DGF发生及改善预后中的潜在治疗价值。

Delayed graft function (DGF) is the most common complication in the early stages of renal transplantation with donor after citizen’s death(DCD). Ischemia/reperfusion (I/R) injury is the major reason of DGF, and renal tubular injury plays the central role. Recent researches and previous work of our group had shown that NK cells are involved in the DGF pathogenesis; NK cells were co-cultured with oxygen-glucose deprivation (OGD) treated HK2 cells, and increases exosomal secretion and with significant miR-21 increase. The miR-21 signal was enhanced in the co-cultured HK2 nuclear region, accompanied by decreased autophagy and increased apoptosis. It is reported that the nuclear entering microRNAs participate gene transcription and modification, while whether miR-21 in nuclear plays the same role is unknown. In this project, the in vitro co-culture model and the in vivo mouse kidney transplantation I/R injury model will introduced, the gene editing, RNA imagination, and RIP-Mass / Seq methods are employed for the purpose to identify the function and the dose-response of NK derived miR-21 in tubular epithelium injury, as well as the targets in nucleus and the underlying mechanism. Furthermore, the target interventions will be performed in vitro and in vivo to evaluate the potential value of this pathway in reducing DGF occurrence and improving prognosis.