Wu Xiangjie, Chen Yiqiong, Chen Suping, Lin Yiping
Department of Pediatrics, School of Medicine, Jinhua Polytechnic, Jinhua, Zhejiang 321007, P.R. China.
Exp Ther Med. 2024 Dec 13;29(2):30. doi: 10.3892/etm.2024.12780. eCollection 2025 Feb.
The therapeutic role of blockade of cysteinyl leukotriene receptor 1 (CysLTR1) in asthma has been previously studied. However, the effect of CysLTR1 blockade on bronchial epithelial cell apoptosis and the nuclear factor erythroid-derived 2-related factor 2 (Nrf2) signaling pathway remains unclear. The present study established an ovalbumin (OVA)-induced asthmatic rat model. Varying doses (1, 4 and 30 mg/kg) of montelukast sodium, a specific CysLTR1 antagonist, were used to inhibit CysLTR1 function in an asthmatic rat model. Reverse transcription-quantitative PCR was used to detect the expression levels of CysLTR1, NAD(P)H quinone oxidoreductase 1 (NQO1) and heme oxygenase 1 (HO-1). CysLTR1 and Nrf2 protein expression levels were determined using western blotting. Immunofluorescence assays were used to evaluate the relative fluorescence intensity of Nrf2 in rat lung tissues. Lung tissue histology was assessed through hematoxylin & eosin, alcian blue and periodic acid-Schiff and Masson's trichrome staining assays. The levels of IL-17, IL-4, serum IgE and the reduced/oxidized glutathione ratio were determined using ELISA assay kits. The number of inflammatory cells was analyzed using Wright-Giemsa staining. Bronchial epithelial cell apoptosis was measured using a TUNEL assay. The results indicated that OVA-induced inflammatory responses and increased eosinophil, lymphocyte and macrophage counts were significantly attenuated following blockade of CysLTR1. Downregulated expression of antioxidant genes NQO1 and HO-1 and the reduced GSH/GSSG ratio caused by OVA challenge were restored by blockade of CysLTR1. Additionally, CysLTR1 blockade also reduced collagen deposition, suppressed goblet cell hyperplasia and inhibited bronchial epithelial cell apoptosis in a rat model of asthma. Furthermore, it was demonstrated that the blockade of CysLTR1 could significantly increase Nrf2 expression. In conclusion, the blockade of CysLTR1 could alleviate asthma in an OVA-induced rat model by inhibiting bronchial epithelial cell apoptosis and activating the Nrf2 signaling pathway. These data may potentially provide a theoretical basis for future asthma therapy in a clinical setting.
此前已对阻断半胱氨酰白三烯受体1（CysLTR1）在哮喘治疗中的作用进行了研究。然而，CysLTR1阻断对支气管上皮细胞凋亡及核因子红细胞2相关因子2（Nrf2）信号通路的影响仍不清楚。本研究建立了卵清蛋白（OVA）诱导的哮喘大鼠模型。使用不同剂量（1、4和30mg/kg）的特异性CysLTR1拮抗剂孟鲁司特钠，在哮喘大鼠模型中抑制CysLTR1功能。采用逆转录定量PCR检测CysLTR1、NAD（P）H醌氧化还原酶1（NQO1）和血红素加氧酶1（HO-1）的表达水平。使用蛋白质印迹法测定CysLTR1和Nrf2蛋白表达水平。采用免疫荧光分析评估大鼠肺组织中Nrf2的相对荧光强度。通过苏木精-伊红、阿尔辛蓝和过碘酸-希夫以及Masson三色染色分析评估肺组织组织学。使用ELISA试剂盒测定IL-17、IL-4、血清IgE水平及还原型/氧化型谷胱甘肽比值。使用瑞氏-吉姆萨染色分析炎症细胞数量。采用TUNEL分析检测支气管上皮细胞凋亡。结果表明，阻断CysLTR1后，OVA诱导的炎症反应及嗜酸性粒细胞、淋巴细胞和巨噬细胞计数增加均显著减轻。OVA攻击导致的抗氧化基因NQO1和HO-1表达下调及GSH/GSSG比值降低，通过阻断CysLTR1得以恢复。此外，在哮喘大鼠模型中，阻断CysLTR1还减少了胶原沉积，抑制了杯状细胞增生并抑制了支气管上皮细胞凋亡。此外，还证明阻断CysLTR1可显著增加Nrf2表达。总之，阻断CysLTR1可通过抑制支气管上皮细胞凋亡和激活Nrf2信号通路，减轻OVA诱导的大鼠模型中的哮喘。这些数据可能为未来临床环境中的哮喘治疗提供理论依据。
