降钙素基因相关肽中和抗体在鼠模型过敏性鼻炎治疗中的作用

文献信息

中文摘要

引言： 本研究旨在表征过敏性鼻炎（AR）患者中降钙素基因相关肽（CGRP）的表达，并在鼠模型中探讨CGRP干预对AR症状、鼻黏膜炎症及2型固有淋巴细胞（ILC2s）数量的影响。
方法： 收集20例AR患者和20例对照受试者的外周血和下鼻甲黏膜样本。分别采用酶联免疫吸附试验（ELISA）和免疫组织化学（IHC）检测血清和鼻黏膜中的CGRP水平。将32只C57BL/6J小鼠随机分为四组：对照组、AR组、AR+同型IgG组和AR+抗CGRP组。AR组小鼠用卵清蛋白（OVA）致敏并鼻内激发。在激发阶段，AR+同型IgG组和AR+抗CGRP组小鼠分别接受同型IgG和AR+抗CGRP治疗，而对照组小鼠仅接受磷酸盐缓冲液（PBS）。根据擦鼻和打喷嚏的频率以及流涕的严重程度评估AR相关鼻症状。采用苏木精-伊红（HE）和高碘酸-希夫（PAS）染色评估鼻黏膜的组织病理学变化及杯状细胞数量增加。采用IHC检测IL-1β、IL-6、TNF-α和CGRP的表达水平。进行双重免疫荧光染色以定量鼻黏膜中的ILC2s。采用蛋白质印迹（WB）测定MAPK通路关键蛋白（包括JNK、ERK1/2和p38）的磷酸化水平。
结果： AR患者鼻黏膜和血清中的CGRP表达水平显著高于对照组。血清CGRP水平与视觉模拟量表（VAS）评分呈正相关，鼻黏膜CGRP表达与炎症细胞数量呈正相关。与AR组和AR+同型IgG组相比，抗CGRP组小鼠的鼻症状显著减轻，鼻黏膜炎症细胞浸润明显减少。此外，鼻黏膜中IL-1β、IL-6、TNF-α和CGRP的表达水平降低，ILC2s数量减少，p-JNK、p-ERK1/2和p-p38的磷酸化水平显著降低。
结论： CGRP可能作为AR的潜在治疗靶点。CGRP中和抗体可以减轻AR症状、抑制炎症反应并调节ILC2s数量。

英文摘要

INTRODUCTION: This study aimed to characterize the expression of calcitonin gene-related peptide (CGRP) in patients with allergic rhinitis (AR) and to investigate the effects of CGRP intervention on AR symptoms, nasal mucosal inflammation, and the number of group 2 innate lymphoid cells (ILC2s) in a murine model.
METHODS: Peripheral blood and inferior turbinate mucosal samples were collected from 20 patients with AR and 20 control subjects. CGRP levels in serum and nasal mucosa were measured by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry (IHC), respectively. Thirty-two C57BL/6J mice were randomly assigned to four groups: Control, AR, AR + isotype IgG, and AR + anti-CGRP. Mice in the AR group were sensitized and intranasally challenged with ovalbumin (OVA). During the challenge phase, mice in the AR + isotype IgG group and the AR + anti-CGRP group received isotype IgG and AR + anti-CGRP treatment, respectively, whereas mice in the Control group received phosphate-buffered saline (PBS) only. AR-related nasal symptoms were evaluated based on the frequency of nose rubbing and sneezing, as well as the severity of rhinorrhea. Hematoxylin-eosin (HE) and periodic acid-Schiff (PAS) staining were used to assess histopathological changes in the nasal mucosa and the increased number of goblet cells. IHC was used to detect the expression levels of IL-1β, IL-6, TNF-α, and CGRP. Double immunofluorescence staining was performed to quantify ILC2s in the nasal mucosa. Western blotting (WB) was used to determine the phosphorylation levels of the key proteins in the MAPK pathway, including JNK, ERK1/2, and p38.
RESULTS: CGRP expression levels in the nasal mucosa and serum of patients with AR were significantly higher than those in the control group. Serum CGRP levels were positively correlated with visual analog scale (VAS) scores, and nasal mucosal CGRP expression was positively correlated with the number of inflammatory cells. Compared with the AR group and the AR + isotype IgG group, mice in the Anti-CGRP group showed significantly alleviated nasal symptoms and markedly reduced inflammatory cell infiltration in the nasal mucosa. In addition, the expression levels of IL-1β, IL-6, TNF-α, and CGRP in the nasal mucosa were decreased, the number of ILC2s was reduced, and the phosphorylation levels of p-JNK, p-ERK1/2, and p-p38 were significantly lowered.
CONCLUSION: CGRP may serve as a potential therapeutic target for AR. CGRP-neutralizing antibody can alleviate AR symptoms, suppress inflammatory responses, and regulate the number of ILC2s.