肝静脉闭塞病是造血干细胞移植后常见并发症之一，肝窦内皮细胞及肝细胞损伤是肝静脉闭塞病的病理生理基础。NF-κB信号是肝细胞修复的重要信号通路之一，IKKβ是NF-κB信号通路中主要的分子。长片段非编码RNA（lncRNA）是基因表达调控的新切入点，我们发现lncRNA uc009ldv.1与IKKβ同源；造血干细胞移植小鼠肝损伤修复模型中肝细胞内uc009ldv.1与IKKβ同步上升，二者呈正相关。提示该lncRNA可能是NF-kB的正性调节因子，可调控NF-kB表达影响肝损伤的修复。本课题拟采用肝细胞损伤体外和体内实验，明确uc009ldv.1在损伤肝细胞修复中的作用，从DNA甲基化、组蛋白修饰及蛋白翻译等方面探索其调控NF-κB表达机制，拓宽人们对NF-kB信号通路的认识，进一步阐明肝静脉闭塞病中肝修复的机理，为临床多种相关肝损伤疾病的治疗提供新的实验及理论依据。

Hepatic veno-occlusive disease is a common complication after hematopoietic stem cell transplantation, and the damage of liver sinusoidal endothelial cells and hepatocyte is the pathophysiological basis of hepatic veno-occlusive disease. NF-κB signaling is one of the important signaling pathways in the repairation of the hepatocyte, IKKβ is the main molecular of NF-κB signaling pathway. The long non-coding RNA (lncRNA) is a new entry point of the gene expression regulation, we found lncRNA uc009ldv.1 and IKKβ display sequence similarity. In liver injury and repair model of hematopoietic stem cell transplantation, uc009ldv.1 and IKKβ in mice hepatocyte were positively correlated. The results suggest that lncRNA uc009ldv.1 is likely a positive regulator of NF-kB, which can regulate the expression of NF-kB and effect the repair of the damage hepatocyte. In the paper, we intend to clear the roles of the uc009ldv.1 in the repar of the hepatocyte damage from the DNA methylation, histone modification and protein expression to explore the mechanism of NF-κB expression, broaden the understanding of NF-κB signaling pathway, further elucidate the mechanism of liver repair of hepatic veno-occlusive disease, and to provide new experimental and theoretical basis for the clinical treatment of various diseases related to liver damage.

本研究通过构建慢病毒lncRNAuc0091dv.1载体，感染肝细胞株，建立肝损伤模型，通过Edu增殖实验、流式细胞术、Wsternblot、qPCR等技术检测lncRNAuc0091dv.1对损伤肝细胞的影响，明确了lncRNAuc0091dv.1/ NF-κB对肝细胞损伤修复的调控作用。具体结果如下：1）成功构建了慢病毒Uc0091dv.1载体，过表达Uc0091dv.1慢病毒成功感染肝细胞株，感染效率超过95%，成功转入小鼠肝细胞内；2）通过给予移植预处理药物白消安损伤肝脏，建立肝细胞损伤模型，明确lncRNAuc0091dv.1对损伤肝细胞的影响。3）通过增殖及凋亡试验，发现过表达Uc0091dv.1对损伤肝细胞的增殖率较对照组升高；过表达Uc0091dv.1对损伤肝细胞的凋亡率较对照组降低。说明lncRNAuc0091dv.1可促进肝细胞增生，减轻肝细胞损伤，同时减缓肝细胞的凋亡，结合前期研究，骨髓移植小鼠肝损伤修复模型中肝细胞内uc009dv.1跟IKKβ同步上升，二者呈正相关，且两者具有同源性，提示lncRNAuc0091dv.1是NF-kB的正性调节子，可调控NF-kB表达，参与损伤肝细胞的增殖、修复与凋亡，本课题的研究成果可为HVOD的预防和治疗提供新的思路。