Abstract：
The present study aims to elucidate the possible involvement of H19 in primary graft dysfunction (PGD) following lung transplantation (LT) and the underlying mechanism. The transcriptome data were obtained through high-throughput sequencing analysis, and the differential long noncoding RNAs and messenger RNAs were screened for coexpression analysis. The interaction among H19, KLF5 and CCL28 was analyzed. A hypoxia-induced human pulmonary microvascular endothelial cell injury model was established, in which H19 was knocked down to elucidate its effect on the lung function, inflammatory response, and cell apoptosis. An orthotopic left LT model was constructed for in vivo mechanistic validation. High-throughput transcriptome sequencing analysis revealed the involvement of the H19/KLF5/CCL28 signaling axis in PGD. Silencing of H19 reduced inflammatory response and thus improved PGD. CCL28 secreted by human pulmonary microvascular endothelial cells after LT recruited neutrophils and macrophages. Mechanistic investigations indicated that H19 augmented the expression of CCL28 by binding to the transcription factor KLF5. Abundant expression of CCL28 reversed the alleviating effect of H19 silencing on PGD. In conclusion, the results point out that H19 exerts a promoting effect on PGD through increasing KLF5 expression and the subsequent CCL28 expression. Our study provides a novel insight into the mechanism of action of H19.
摘要：
本研究旨在阐明H19在肺移植（LT）后原发性移植物功能障碍（PGD）中的可能参与及其潜在机制。通过高通量测序分析获得转录组数据，并筛选差异长链非编码RNA和信使RNA进行共表达分析。分析了H19、KLF5和CCL28之间的相互作用。建立缺氧诱导人肺微血管内皮细胞损伤模型,其中H19被击倒以阐明其对肺功能的影响，炎症反应，和细胞凋亡。构建原位左LT模型用于体内机制验证。高通量转录组测序分析显示H19/KLF5/CCL28信号轴参与PGD。H19的沉默减少了炎症反应，从而改善了PGD。人肺微血管内皮细胞在LT后分泌的CCL28募集中性粒细胞和巨噬细胞。机制研究表明，H19通过与转录因子KLF5结合来增强CCL28的表达。CCL28的大量表达逆转了H19沉默对PGD的缓解作用。总之,结果表明，H19通过增加KLF5的表达和随后的CCL28的表达对PGD具有促进作用。我们的研究为H19的作用机制提供了新的见解。
