Xu Mengrou, Hu Bin, Chen Jiarui, Zhao Limin, Wang Jing, Li Xiaoyan
Department of Otorhinolaryngology Head and Neck Surgery, Shanghai Children's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Department of Otorhinolaryngology Head and Neck Surgery, Changhai Hospital Affiliated with the Second Military Medical University of PLA, Shanghai, China.
Transl Pediatr. 2023 Sep 18;12(9):1634-1645. doi: 10.21037/tp-23-118. Epub 2023 Sep 14.
Laryngotracheal stenosis (LTS) is a life-threatening disease that commonly results in airway obstruction in children. Traditional treatments such as laryngotracheal reconstruction and balloon dilation all have the risk of laryngotracheal restenosis. It is of great importance to spare patients the morbidity of LTS and risks of restenosis associated with these treatments. Laboratory and clinical trials have focused on fibrosis, the crucial pathological process of LTS. This study was undertaken to investigate the function of CXC chemokine receptor-7 () in the fibroblasts derived from LTS.
RNA sequencing was performed on acquired human LTS and normal trachea tissues to analyze differentially expressed genes. Fibroblasts from LTS and normal trachea tissues were isolated and cultured. knockdown was performed using specific small interfering RNAs (siRNAs) and activated by CXCR7 agonist VUF11207. The assessment of cell proliferation and migration was conducted using EdU proliferation, wound healing, and transwell assays. The assessment of cell proliferation and migration was conducted using EdU proliferation, wound healing, and transwell assays. The expressions of , and NF-κB signaling pathway were analyzed by quantitative polymerase chain reaction (qPCR), western blotting, immunohistochemistry, and immunofluorescence.
RNA sequencing showed that was among the most differentially expressed genes. LTS had an increased CXCR7 expression but decreased E-cadherin expression . CXCR7 agonist stimulated the migration of LTS derived fibroblasts significantly , with no significant influence on the cell proliferation and apoptosis. CXCR7 agonist inhibited the expression of by activating the NF-κB signaling pathway. The effects of on cell migration and expression were blocked by siRNA.
LTS had an increased expression but decreased expression. activation inhibited expression by NF-κB signaling pathway and thereby promoted the migration of LTS derived fibroblasts.
喉气管狭窄（LTS）是一种危及生命的疾病，在儿童中常导致气道阻塞。喉气管重建和球囊扩张等传统治疗方法均有喉气管再狭窄的风险。避免患者发生LTS的发病率以及与这些治疗相关的再狭窄风险非常重要。实验室和临床试验都聚焦于纤维化，这是LTS的关键病理过程。本研究旨在探讨CXC趋化因子受体7（CXCR7）在LTS来源的成纤维细胞中的作用。
对获取的人LTS组织和正常气管组织进行RNA测序，以分析差异表达基因。分离并培养LTS组织和正常气管组织的成纤维细胞。使用特异性小干扰RNA（siRNAs）进行CXCR7敲低，并由CXCR7激动剂VUF11207激活。采用EdU增殖、伤口愈合和Transwell实验评估细胞增殖和迁移。通过定量聚合酶链反应（qPCR）、蛋白质免疫印迹法、免疫组织化学和免疫荧光分析CXCR7、E-钙黏蛋白（E-cadherin）和核因子κB（NF-κB）信号通路的表达。
RNA测序显示CXCR7是差异表达最显著的基因之一。LTS中CXCR7表达增加，但E-钙黏蛋白表达降低。CXCR7激动剂显著刺激LTS来源的成纤维细胞迁移，对细胞增殖和凋亡无显著影响。CXCR7激动剂通过激活NF-κB信号通路抑制E-钙黏蛋白的表达。CXCR7 siRNA阻断了CXCR7对细胞迁移和E-钙黏蛋白表达的影响。
LTS中CXCR7表达增加，但E-钙黏蛋白表达降低。CXCR7激活通过NF-κB信号通路抑制E-钙黏蛋白表达，从而促进LTS来源的成纤维细胞迁移。
