大鼠喉部肌肉发育过程中及神经损伤后HGF和Met的表达

文献信息

中文摘要

目的： 喉返神经损伤导致强烈但联带的神经再生，这凸显了更好地表征引导正确初级神经支配的神经营养因子的必要性。肝细胞生长因子（HGF）是一种趋化因子，通过其受体Met赋予神经元存活。本研究使用RNAScope检测发育中大鼠喉内肌（ILMs）中的HGF和Met，并使用RNASeq检测发育过程中及损伤后相关基因的表达。
方法： 使用QuPath定量大鼠胚胎后环杓肌（PCA）、外侧甲杓肌（LTA）和内侧甲杓肌（MTA）中HGF和Met RNA表达为 puncta/细胞。从发育过程中及成年神经损伤后的PCA、LTA和MTA中提取RNA，并进行RNASeq。对HGF相关基因进行生物信息学分析。在R中进行单因素和双因素方差分析及事后Tukey检验。
结果： HGF puncta/细胞随年龄（p = 0.007）和ILM（p = 0.0257）在发育过程中显著变化，PCA和LTA中表达随年龄降低（p = 0.042和p = 0.048）。Met在PCA（p = 0.002）和MTA（p = 0.011）中随年龄增加，在LTA中降低（p = 0.0095）。RNASeq显示HGF-Met通路中几个相关蛋白的显著上调和下调，以及损伤后Met/HGF表达的变化，主要在PCA中。
结论： HGF和Met在发育中大鼠ILMs中表达，在初级神经支配的关键点具有时间变化，损伤后在PCA中上调。这些发现表明HGF-Met信号通路在初级喉神经支配中发挥作用，神经损伤后可能失调。
证据等级： 不适用。

英文摘要

OBJECTIVES: Injury to the recurrent laryngeal nerve results in robust but synkinetic nerve regrowth, highlighting the need to better characterize neurotrophic factors guiding correct primary innervation. Hepatocyte growth factor (HGF) is a chemoattractant that confers neuronal survival through its receptor Met. This study examined HGF and Met within the developing rat intrinsic laryngeal muscles (ILMs) using RNAScope and associated gene expression during development and after injury using RNASeq.
METHODS: HGF and Met RNA expression were quantified as puncta/cell using QuPath within the posterior cricoarytenoid (PCA), lateral thyroarytenoid (LTA), and medial thyroarytenoid (MTA) of rat embryos. RNA was extracted from PCA, LTA, and MTA during development and following nerve injury in adults, and RNASeq was performed. Bioinformatic analysis was conducted for HGF-associated genes. One- and two-way ANOVA analyses and post hoc Tukey's tests were performed in R.
RESULTS: HGF puncta/cell varied significantly throughout development by age (p = 0.007) and ILM (p = 0.0257), with expression decreasing with age in both the PCA and LTA (p = 0.042 and p = 0.048). Met increased with age within the PCA (p = 0.002) and MTA (p = 0.011), and decreased in the LTA (p = 0.0095). RNASeq demonstrated significant up- and downregulation of several associated proteins along the HGF-Met pathway and variation in Met/HGF expression post-injury, primarily in the PCA.
CONCLUSIONS: HGF and Met were expressed in the developing rat ILMs with temporal variation at key points in primary innervation, with post-injury upregulation in the PCA. These findings suggest a role for the HGF-Met signaling pathway in primary laryngeal innervation with potential dysregulation after nerve injury.
LEVEL OF EVIDENCE: N/A.