降钙素基因相关肽调节特定干扰素刺激基因以抑制干眼病中角膜上皮细胞的凋亡。

Calcitonin Gene-Related Peptide Regulates Specific Interferon-Stimulating Genes to Inhibit Apoptosis of Corneal Epithelial Cells in Dry Eye Disease.

作者信息

Hong Xiaoping, Ding Fadian, Zhang Ling, Lv Runhua, Chen Jiaxin, Gao YingYing

机构信息

Department of Ophthalmology, The Second Affiliated Hospital of Fujian Medical University; Fujian Medical University, Quanzhou, People's Republic of China.

Hepatopancreatobiliary Surgery Department, the First Affiliated Hospital of Fujian Medical University, Fuzhou, People's Republic of China.

出版信息

Invest Ophthalmol Vis Sci. 2025 Jul 1;66(9):65. doi: 10.1167/iovs.66.9.65.

DOI:10.1167/iovs.66.9.65

PMID:40728362

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12315935/

Abstract

PURPOSE

Calcitonin gene-related peptide (CGRP) has demonstrated the potential to treat dry eye disease (DED), although its mechanism is not well-understood. This study investigated the effect of CGRP on apoptosis of corneal epithelial cells in a model of DED.

METHODS

Tear samples were collected from patients with DED. Additionally, samples were collected from a rat model of DED and a CGRP overexpression human corneal epithelial (HCET) model. The concentrations of CGRP and IFN-γ in tears were detected by ELISA and analyzed in combination with clinical data. The apoptosis level of corneal epithelial cells was evaluated by Western blot, qRT-PCR, TUNEL staining, and flow cytometry. RNA-seq screening and validation of CGRP target proteins in corneal epithelial cells was also performed.

RESULTS

The findings of this study demonstrated a significant and positive correlation between CGRP expressed in tears of patients with DED and multiple clinical indicators of DED (P < 0.05). CGRP expression was similarly increased in the DED animal model. Exogenous CGRP peptides were observed to significantly inhibit the apoptosis of corneal tissue (P < 0.05). The CGRP overexpressing group similarly showed decreased levels of apoptosis in the corneal tissue (P < 0.05). Reduced apoptosis of corneal tissues was associated with the IFN-γ/JAK2/STAT1 pathway. RNA-seq suggested that CGRP(8-37) concurrently increased apoptosis, inhibited the expression of specific interferon-stimulating genes (SISGs), and was related to the activation of IFN-γ/JAK2/STAT1 (P < 0.05).

CONCLUSIONS

High CGRP concentration in tears of patients with DED demonstrated a significant correlation with severity of clinical symptoms. Increased CGRP was associated with reduced apoptosis of corneal epithelial cells in an animal model of DED. Molecular evaluation identified inhibition of SISGs and activation of IFN-γ/JAK2/STAT1 as CGRP-related mechanisms potentially mediating reduced apoptosis.

摘要

目的

降钙素基因相关肽（CGRP）已显示出治疗干眼病（DED）的潜力，但其机制尚不清楚。本研究在DED模型中研究了CGRP对角膜上皮细胞凋亡的影响。

方法

收集DED患者的泪液样本。此外，从DED大鼠模型和CGRP过表达人角膜上皮（HCET）模型中收集样本。通过ELISA检测泪液中CGRP和IFN-γ的浓度，并结合临床数据进行分析。通过蛋白质免疫印迹、qRT-PCR、TUNEL染色和流式细胞术评估角膜上皮细胞的凋亡水平。还进行了RNA测序筛选和验证角膜上皮细胞中CGRP靶蛋白。

结果

本研究结果表明，DED患者泪液中表达的CGRP与DED的多个临床指标之间存在显著正相关（P<0.05）。在DED动物模型中，CGRP表达同样增加。观察到外源性CGRP肽可显著抑制角膜组织的凋亡（P<0.05）。CGRP过表达组的角膜组织凋亡水平同样降低（P<0.05）。角膜组织凋亡减少与IFN-γ/JAK2/STAT1通路有关。RNA测序表明，CGRP(8-37)同时增加凋亡，抑制特定干扰素刺激基因（SISGs）的表达，并与IFN-γ/JAK2/STAT1的激活有关（P<0.05）。