Department of Otorhinolaryngology-Head and Neck Surgery, Lu'an Hospital of Anhui Medical University, Lu'an People's Hospital of Anhui Province, Lu'an, 237000, Anhui, China.
Translational Medicine Center, Zhangjiagang Hospital Affiliated to Soochow University, Suzhou, 215000, China.
Eur Arch Otorhinolaryngol. 2023 Feb;280(2):703-711. doi: 10.1007/s00405-022-07538-w. Epub 2022 Jul 13.
To investigate the therapeutic effect of Bicalutamide, an androgen receptor antagonist on the onset and development of allergic rhinitis in an animal model.
40 male BALB/c mice were randomly divided into five groups (eight mice per group). Aluminum hydroxide powder was used as an adjuvant, combined with Ovalbumin (OVA) to establish the mouse model of allergic rhinitis via ultrasonic nebulization of OVA to stimulate the nasal cavity. Mice in Bica#1 group were intraperitoneally injected with 0.02 mg Bicalutamide/0.5 ml of normal saline daily for 7 consecutive days; mice in Bica#2 group were administered 0.02 mg Bicalutamide/0.5 ml of normal saline via intraperitoneal injection for 5 consecutive days, and then the same amount of normal saline was injected intraperitoneally for 2 consecutive days. Enzyme-linked immunosorbent assay was adopted to detect the serological levels of IgE, IL-4, and IL-6 production. Eosinophil infiltration was observed under microscope after hematoxylin and eosin staining of nasal mucosa. Quantitative PCR and Western blot were employed for determination of histamine receptors mRNA expression and PI3K/PKB associated protein levels, respectively.
Histological analysis shown that allergic lesion was induced after OVA sensitization. Intraperitoneal injection with 0.02 mg Bicalutamide daily for 7 consecutive days significantly reduced the allergic lesion; however, mice injected with the same amount of normal saline at the same time demonstrated no allergic rhinitis symptoms. In addition, there was a significant reduction in eosinophils number in Bicalutamide treated mice (n = 8) compared to the OVA group (n = 8) (OVA: 19.6 ± 5.3 vs. Bica#1: 7.7 ± 0.8 vs. Bica#2: 9.4 ± 1.2, both p < 0.01). Furthermore, ELISA results revealed that the serological levels of IgE (OVA: 17.3 ± 1.7 µg/ml vs. Bica#1: 9.2 ± 0.6 vs. Bica#2: 10.4 ± 2.3, both p < 0.05), IL-4 (OVA: 164.3 ± 5.1 pg/ml vs. Bica#1: 110.2 ± 3.1 vs. Bica#2: 115.3 ± 4.1, both p < 0.05) and IL-6 (OVA: 167.3 ± 3.7 pg/ml vs. Bica#1: 117.5 ± 6.5 vs. Bica#2: 114.8 ± 2.4, both p < 0.05) were significantly decreased after two different dosage of Bicalutamide treatment. Similarly, histamine receptors in mast cells were significantly reduced after two different dosage of Bicalutamide treatment. More importantly, p-PKB protein was notably reduced after two different dosage of Bicalutamide treatment compared to the OVA group, mTOR protein levels were also down regulated after two different dosage of Bicalutamide treatment.
Our data demonstrated that androgen receptor antagonist Bicalutamide can significantly alleviate allergic rhinitis lesion in the animal model. PI3K/PKB activity in mast cells was suppressed after Bicalutamide injection. Our results provide important implication in allergic rhinitis prevention and treatment.
研究雄激素受体拮抗剂比卡鲁胺对变应性鼻炎发病和发展的治疗作用。
将 40 只雄性 BALB/c 小鼠随机分为五组（每组 8 只）。采用氢氧化铝粉末作为佐剂，联合卵清蛋白（OVA）经超声雾化刺激鼻腔，建立小鼠变应性鼻炎模型。Bica#1 组小鼠每日腹腔注射 0.02 mg 比卡鲁胺/0.5 ml 生理盐水，连续 7 天；Bica#2 组小鼠连续 5 天腹腔注射 0.02 mg 比卡鲁胺/0.5 ml 生理盐水，然后连续 2 天腹腔注射等量生理盐水。采用酶联免疫吸附试验检测血清 IgE、IL-4 和 IL-6 水平。苏木精-伊红染色后在显微镜下观察鼻黏膜嗜酸性粒细胞浸润情况。采用定量 PCR 和 Western blot 分别检测组氨酸受体 mRNA 表达和 PI3K/PKB 相关蛋白水平。
OVA 致敏后可诱导过敏病变。连续 7 天每天腹腔注射 0.02 mg 比卡鲁胺可显著减轻过敏病变；但同时注射等量生理盐水的小鼠则无变应性鼻炎症状。比卡鲁胺治疗组（n=8）与 OVA 组（n=8）相比，嗜酸性粒细胞数量明显减少（OVA：19.6±5.3 比 Bica#1：7.7±0.8 比 Bica#2：9.4±1.2，均 p<0.01）。此外，ELISA 结果显示，血清 IgE（OVA：17.3±1.7 μg/ml 比 Bica#1：9.2±0.6 比 Bica#2：10.4±2.3，均 p<0.05）、IL-4（OVA：164.3±5.1 pg/ml 比 Bica#1：110.2±3.1 比 Bica#2：115.3±4.1，均 p<0.05）和 IL-6（OVA：167.3±3.7 pg/ml 比 Bica#1：117.5±6.5 比 Bica#2：114.8±2.4，均 p<0.05）水平在两种不同剂量的比卡鲁胺治疗后均显著降低。同样，两种不同剂量的比卡鲁胺治疗后，肥大细胞中的组氨酸受体也明显减少。更重要的是，与 OVA 组相比，两种不同剂量的比卡鲁胺治疗后 p-PKB 蛋白明显减少，mTOR 蛋白水平也下调。
我们的数据表明，雄激素受体拮抗剂比卡鲁胺可显著减轻动物模型中的变应性鼻炎病变。比卡鲁胺注射后，肥大细胞中的 PI3K/PKB 活性受到抑制。我们的结果为变应性鼻炎的预防和治疗提供了重要依据。
