Effects of lentivirus-mediated CCR3 RNA interference on the function of mast cells of allergic rhinitis in mice
Effects of lentivirus-mediated CCR3 RNA interference on the function of mast cells of allergic rhinitis in mice
复制标题
慢病毒介导的CCR3 RNA干扰对小鼠变应性鼻炎肥大细胞功能的影响
DOI：
10.1016/j.intimp.2019.106011
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发表时间：
2020-01-01
影响因子：
5.6
通讯作者：
Zhu, Xinhua
中科院分区：
文献类型：
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作者：
Wu, Shuhong;Tang, Siyi;Zhu, Xinhua
嗜酸性粒细胞、肥大细胞和Th 2细胞表达的CC趋化因子受体3（CCR 3）与变态反应性疾病密切相关。本研究的目的是探讨CCR 3沉默与短发夹RNA（shRNA）由慢病毒载体提供是否可以影响肥大细胞在小鼠模型的过敏性鼻炎（AR）在体内的功能。建立卵清蛋白（OVA）诱导的小鼠变应性鼻炎（AR）模型，将BALB/c小鼠分为正常对照组、AR组、对照组和慢病毒CCR 3-shRNA治疗组。将表达靶向CCR 3的短发夹RNA（shRNA）的重组慢病毒载体滴入OVA致敏小鼠的鼻腔中。采用实时荧光定量PCR和Western blotting方法观察CCR 3基因对细胞增殖的抑制作用。评估各组小鼠的鼻部症状和OVA特异性IgE。分析骨髓、外周血和鼻粘膜中组胺、类胰蛋白酶和前列腺素D2（PGD 2）的浓度。应用组织学和电镜观察鼻黏膜组织学改变及肥大细胞浸润情况。结果表明，CCR 3shRNA能有效抑制骨髓、外周血和鼻黏膜中CCR 3基因的表达，减轻鼻腔症状，降低OVA特异性IgE水平，减少鼻腔炎性细胞和肥大细胞浸润，减轻鼻黏膜组织病理学改变。此外，CCR 3shRNA干预可降低骨髓、外周血和鼻粘膜中组胺、类胰蛋白酶和PGD 2的水平。提示通过shRNA慢病毒载体抑制CCR 3基因表达，可有效抑制局部组织肥大细胞的迁移、浸润和脱颗粒，减轻变应性鼻炎小鼠的炎症反应。
The CC chemokine receptor 3 (CCR3) expressed by eosinophils, mast cells and Th2 cells is closely related to allergic diseases. The objective of this study was to explore whether silencing of CCR3 with short hairpin RNAs (shRNAs) delivered by a lentiviral vector could impact the function of mast cells in a murine model of allergic rhinitis (AR) in vivo. The murine model of allergic rhinitis (AR) inducing by ovalbumin (OVA) was constructed, and the BALB/c mice were divided into normal control group, AR group, controlshRNA treated group and lentiviral CCR3-shRNA treated group. The recombinant lentivirus vectors which express a short hairpin RNA (shRNA) targeting the CCR3 were dropped into the nasal cavity of OVA-sensitized mice before the challenges. Real-time fluorescence quantitative PCR and western blotting were performed to observe inhibitory effect of CCR3 gene. Nasal symptoms of mice and OVA-specific IgE in each group were assessed. Concentrations of histamine, tryptase and Prostaglandin D2 (PGD2) in bone marrow, peripheral blood and nasal mucosa were analyzed. Furthermore, histological analysis and electron microscopy analysis were applied to detect the histology changes of nasal mucosa and the infiltration of mast cells in nasal mucosa. The results showed that administration of CCR3shRNA could effectively inhibit the expression of the CCR3 gene in bone marrow, peripheral blood and nasal mucosa, which reduced the nasal symptoms, the level of OVA-specific IgE, the inflammatory cells and mast cells infiltration into nasal cavity, and relieved the histopathological changes of nasal mucosa. In addition, intervention of CCR3shRNA could reduce the levels of the histamine, tryptase and PGD2 in bone marrow, peripheral blood and nasal mucosa. These results suggest that inhibition of CCR3 gene expression by shRNAs lentiviral vectors can effectively attenuate migration, infiltration and degranulation of mast cells in local tissues and alleviate the inflammation of allergic rhinitis mice.