Department of Pediatrics, South Campus, Renji Hospital, Shanghai Jiao Tong University School of Medicine, No.2000, Jiangyue Road, Pujiang, Minhang District, Shanghai, 201112, China.
Department of Pediatrics, South Campus, Renji Hospital, Shanghai Jiao Tong University School of Medicine, No.2000, Jiangyue Road, Pujiang, Minhang District, Shanghai, 201112, China.
Pulm Pharmacol Ther. 2020 Oct;64:101963. doi: 10.1016/j.pupt.2020.101963. Epub 2020 Oct 6.
Study found that glucocorticoids, as first-line treatments for asthma, fails to prevent asthma recurrence. Orosomucoid-like (ORMDL) 3 is associated to childhood asthma onset and involved in the inflammation and repair of airway epithelium. We explored the functional role of ORMDL3 in glucocorticoid treatment for childhood asthma.
Mice were sensitized with Ovalbumin (OVA) and treated with Dexamethasone (Dex), followed by OVA challenge to establish a mouse model of asthma. Histopathological changes in lung tissues were observed by hematoxylin-eosin and masson staining. Human bronchial epithelial (16HBE-14°) cells were transfected with ORMDL3 overexpression plasmid and siRNA-interleukin (IL)-33 alone or in combination, followed by Dex. Cell viability was measured by MTT assay. Cell migration was evaluated by wound healing assay. The expressions of E-cadherin and Vimentin and the activation of NF-κB and MAPK/ERK in 16HBE-14° cells were assessed by Western blot. The expressions of ORMDL3 and IL-33 in lung tissues and 16HBE-14° cells were analyzed by qRT-PCR or Western blot.
Dex treatment alleviated the histopathological abnormality and reversed the overexpressions of ORMDL3 and IL-33 in the lung tissues of asthmatic mice. Overexpressed ORMDL3 enhanced migration and viability, decreased E-cadherin level, increased the levels of IL-33 and Vimentin, and promoted the phosphorylation of NF-κB and MAPK/ERK in Dex-treated 16HBE-14° cells, thus reversing the effect of Dex treatment. However, siRNA-IL-33 inhibited viability and migration, increased E-cadherin level, decreased Vimentin level, and suppressed the phosphorylation of NF-κB and MAPK/ERK, thus reversing the effect of overexpressed ORMDL3 in Dex-treated 16HBE-14° cells.
ORMDL3 overexpression helped airway epithelial cellrepairin asthma via regulating IL-33 expression.
研究发现，糖皮质激素作为哮喘的一线治疗药物，无法预防哮喘复发。Orosomucoid-like（ORMDL）3 与儿童哮喘发病有关，参与气道上皮的炎症和修复。我们探讨了 ORMDL3 在糖皮质激素治疗儿童哮喘中的功能作用。
用卵清蛋白（OVA）致敏小鼠，并用地塞米松（Dex）治疗，然后用 OVA 攻击建立哮喘小鼠模型。通过苏木精-伊红和 Masson 染色观察肺组织的组织病理学变化。单独或联合转染 ORMDL3 过表达质粒和 siRNA-白细胞介素（IL）-33 后，用 Dex 处理人支气管上皮（16HBE-14°）细胞。通过 MTT 测定法测量细胞活力。通过划痕愈合试验评估细胞迁移。通过 Western blot 评估 16HBE-14°细胞中 E-钙粘蛋白和波形蛋白的表达以及 NF-κB 和 MAPK/ERK 的激活。通过 qRT-PCR 或 Western blot 分析肺组织和 16HBE-14°细胞中 ORMDL3 和 IL-33 的表达。
Dex 治疗缓解了哮喘小鼠肺部的组织病理学异常，并逆转了哮喘小鼠肺部 ORMDL3 和 IL-33 的过度表达。过表达的 ORMDL3 增强了迁移和活力，降低了 E-钙粘蛋白水平，增加了 IL-33 和波形蛋白水平，并促进了 Dex 处理的 16HBE-14°细胞中 NF-κB 和 MAPK/ERK 的磷酸化，从而逆转了 Dex 治疗的效果。然而，siRNA-IL-33 抑制了活力和迁移，增加了 E-钙粘蛋白水平，降低了波形蛋白水平，并抑制了 Dex 处理的 16HBE-14°细胞中 NF-κB 和 MAPK/ERK 的磷酸化，从而逆转了过表达的 ORMDL3 的作用。
ORMDL3 过表达通过调节 IL-33 的表达有助于哮喘时气道上皮细胞的修复。
