Department of Otorhinolaryngology Head and Neck Surgery, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi 330006, P.R. China.
Mol Med Rep. 2018 Oct;18(4):3780-3790. doi: 10.3892/mmr.2018.9380. Epub 2018 Aug 10.
The present study aimed to investigate the effects of C‑C chemokine receptor type 3 (CCR3) gene knockout on allergic rhinitis (AR) in mice, as well as the underlying molecular mechanisms. Ovalbumin was administrated to CCR3+/+ and CCR3‑/‑ BALB/c mice to establish an AR model. The mice were divided into four groups: i) Normal control (CG), ii) AR model (AR), iii) CCR3 knockout CG (CCR3‑/‑CG) and iv) AR model with CCR3 knockout (CCR3‑/‑AR). Histological sections of nasal mucosae were examined by hematoxylin and eosin staining, which revealed that CCR3 knockout suppressed the invasion of inflammatory cells and relieved the damage of nasal mucosae. Peripheral blood smear and nasal‑washing smears were evaluated by Wright's staining. Eosinophil (EOS) numbers in nasal mucosae, peripheral blood, and nasal washings of the various groups were ranked in the order: AR>CCR3‑/‑AR>CG>CCR3‑/‑. mRNA expression levels of CCR3, EOS peroxidase (EPO), EOS cationic protein (ECP), and major basic protein (MBP) in the peripheral serum and nasal washings were detected by reverse transcription‑polymerase chain reaction. Interferon‑γ (IFN‑γ), interleukin (IL)‑4, IL‑10, and immunoglobulin E (IgE) protein levels in the peripheral serum and nasal washings were investigated by ELISA. CCR3 mRNA expression was not detected in the CCR3‑/‑ and CCR3‑/‑AR groups, whereas expression levels in the AR group were markedly higher compared with expression in the CG group. Compared with the CG‑associated groups (i.e., the CG and CCR3‑/‑CG groups), the levels of EPO, ECP, MBP, IL‑4, and IgE were significantly increased in the AR‑associated groups (that is, R and CCR3‑/‑AR). In addition, the CCR3‑/‑AR group mice produced significantly lower levels of EPO, ECP, MBP, IL‑4 and IgE compared with the AR group, whereas the expression levels of IFN‑γ and IL‑10 were increased. CCR3 gene knockout may alleviate EOS invasion and the inflammatory response in AR model mice by reducing the expression levels of EPO, ECP, MBP, IL‑4, and IgE, and increasing the expression of IL‑10 and IFN‑γ.
本研究旨在探讨 C-趋化因子受体 3（CCR3）基因敲除对小鼠变应性鼻炎（AR）的影响及其潜在的分子机制。卵清蛋白（OVA）用于建立 CCR3+/+和 CCR3-/-BALB/c 小鼠的 AR 模型。将小鼠分为四组：i）正常对照组（CG）、ii）AR 模型组（AR）、iii）CCR3 基因敲除 CG 组（CCR3-/-CG）和 iv）AR 模型 CCR3 基因敲除组（CCR3-/-AR）。通过苏木精-伊红（HE）染色观察鼻黏膜组织学切片，结果显示 CCR3 基因敲除抑制了炎性细胞的浸润，缓解了鼻黏膜的损伤。通过 Wright 染色评估外周血涂片和鼻腔冲洗涂片。对各组鼻黏膜、外周血和鼻腔冲洗液中的嗜酸性粒细胞（EOS）数量进行排序，结果为：AR>CCR3-/-AR>CG>CCR3-/-。通过逆转录-聚合酶链反应（RT-PCR）检测外周血清和鼻腔冲洗液中 CCR3、EOS 过氧化物酶（EPO）、EOS 阳离子蛋白（ECP）和主要碱性蛋白（MBP）的 mRNA 表达水平。通过酶联免疫吸附试验（ELISA）检测外周血清和鼻腔冲洗液中干扰素-γ（IFN-γ）、白细胞介素（IL）-4、IL-10 和免疫球蛋白 E（IgE）蛋白水平。在 CCR3-/-和 CCR3-/-AR 组中未检测到 CCR3 mRNA 的表达，而 AR 组的表达水平明显高于 CG 组。与 CG 相关组（CG 和 CCR3-/-CG 组）相比，AR 相关组（AR 和 CCR3-/-AR 组）的 EPO、ECP、MBP、IL-4 和 IgE 水平显著升高。此外，与 AR 组相比，CCR3-/-AR 组小鼠的 EPO、ECP、MBP、IL-4 和 IgE 表达水平显著降低，而 IFN-γ和 IL-10 的表达水平升高。CCR3 基因敲除可能通过降低 EPO、ECP、MBP、IL-4 和 IgE 的表达水平，增加 IL-10 和 IFN-γ 的表达，从而减轻 AR 模型小鼠 EOS 浸润和炎症反应。
