北京中医药大学学报2024，Vol.47Issue(3)：364-373,10.DOI:10.3969/j.issn.1006-2157.2024.03.011
探讨益气温阳方抑制变应性鼻炎小鼠鼻黏膜细胞自噬及保护ZO-1及occludin的作用
Yiqi Wenyang Prescription inhibits autophagy and protects ZO-1 and occludin in nasal mucosal cells of an allergic rhinitis murine model
摘要
Abstract
Objective We aimed to (i) observe the therapeutic effect of Yiqi Wenyang Prescription (YQWYP, which consists of milkvetch root, tangshen, dried ginger, cassig twig, ephedra, biond magnolia flower-bud, Chinese magnoliavine fruit, earthworm, and liquorice root) on an allergic rhinitis (AR) murine model and the protective effect on the tight junction (TJ) key proteins zonula occludens-1 (ZO-1) and occludin and (ii) elucidate whether YQWYP exerts these effects by inhibiting autophagy in nasal mucosal cells.Methods Thirty C57BL/6 mice were randomly divided into six groups with 5 mice per group: (i) the blank group, (ii) the model group, (iii-v) the YQWYP low-dose, mid-dose, and high-dose groups(7, 14, 28 g/kg), and (vi) the cetirizine group(1.667 mg/kg). The model was established by intraperitoneal injection and nasal stimulation of ovalbumin. 1 hour before the nasal stimulation, mice in the blank group or in the model group were given normal saline by gavage, mice in the YQWYP groups and those in the cetirizine group were given the corresponding medicines solution by gavage. After 7 days, mice behavior was scored. After that, the mice were sacrificed and the nasal mucosa tissues were harvested. The inflammatory reaction of the nasal mucosa was observed by HE staining, the proliferation of goblet cells in the nasal mucosa was observed by PAS staining, the IgE content in nasal lavage fluid was determined by ELISA, the expression levels of ZO-1 and occludin were analyzed by immunohistochemistry, and the expression levels of LC3B, P62, and Beclin1 in the nasal mucosa of the blank group, model group, and YQWYP high-dose group were analyzed by Western blotting.Results After the successful establishment of the AR model, compared with the blank group, the symptom scores of nose scratching, sneezing, and runny nose in the model group were higher, while the total scores was significantly higher(P<0.01). In the model group, the nasal mucosal epithelium was disrupted and destroyed, a large number of inflammatory cells were infiltrated, goblet cells showed hyperplasia, mucus production was increased, mucosal swelling was obvious, the amount of IgE in nasal lavage fluid was increased (P<0.01). The protein expression levels of ZO-1 and occludin were decreased (P<0.05), the protein expression levels of LC3BⅡ/Ⅰ and Beclin1 were increased (P<0.05), P62 protein expression was decreased (P<0.05). After medication intervention, compared with the model group, in the YQWYP high-dose group and the cetirizine group, the total score were reduced (P<0.01), the swelling of the nasal mucosa was reduced, the infiltration of inflammatory cells was inhibited, the number of goblet cells was decreased, the amount of IgE in nasal lavage fluid was reduced(P<0.01), the protein expression of ZO-1 and occludin were increased (P<0.05), the protein levels of LC3BII/I and Beclin1 were decreased (P<0.05), P62 protein expression was increased (P<0.05). Conclusion YQWYP has a good therapeutic effect on AR mice, and its mechanism may be related to the promotion of nasal mucosal repair by inhibiting autophagy of nasal mucosal cells.关键词
益气温阳方/变应性鼻炎/自噬/紧密连接/闭锁连接蛋白1/闭合蛋白/小鼠Key words
Yiqi Wenyang Prescription/allergic rhinitis/autophagy/tight junction/zonula occludens-1/occludin/mice分类
医药卫生引用本文复制引用
顾文哲,史军,严道南,沈政洁..探讨益气温阳方抑制变应性鼻炎小鼠鼻黏膜细胞自噬及保护ZO-1及occludin的作用[J].北京中医药大学学报,2024,47(3):364-373,10.基金项目
国家自然科学基金项目(No.82104939) （No.82104939）
苏州市卫生青年骨干人才"全国导师制"培训项目(No.Qngg2022035) National Natural Science Foundation of China(No.82104939) （No.Qngg2022035）
