不同剂量1,25-(OH)2D3对哮喘小鼠肺内高迁移率族蛋白B1及IL-17表达的影响

Effects of different doses of 1,25-(OH)2D3 on expression of HMGB1 and IL-17 in the lungs of asthmatic mice

摘要:

目的 观察不同剂量1,25-二羟基维生素D₃ [1,25-(OH)₂D₃]对哮喘小鼠肺内高迁移率族蛋白B1(HMGB1)及IL-17表达的影响.方法 将50只BALB/c小鼠随机分为对照组、哮喘组、低剂量组、中剂量组和高剂量组,每组10只.卵清蛋白混合液致敏并雾化吸入建立哮喘小鼠模型,低、中、高剂量组在每次激发前分别按1、4、10 μg/kg给予腹腔注射1,25-(OH)₂D₃混合液,对照组和哮喘组以生理盐水替代.采用苏木精-伊红染色观察小鼠气道结构变化;免疫组化染色观察HMGB1、IL-17蛋白表达的变化,RT-PCR检测HMGB1及IL-17 mRNA水平的表达变化.结果 哮喘组气道壁厚度、HMGB1和IL-17的 mRNA及蛋白表达水平均明显高于对照组(P<0.05);低、中剂量组气道壁厚度、HMGB1和IL-17的 mRNA及蛋白表达水平均明显低于哮喘组,且上述指标在中剂量组均明显低于低剂量组(P<0.05);但高剂量组中气道壁厚度、HMGB1和IL-17的 mRNA及蛋白表达水平均高于哮喘组(P<0.05).结论 HMGB1及IL-17可能参与哮喘气道重塑过程;适量1,25-(OH)₂D₃能改善哮喘小鼠气道重塑,大剂量1,25-(OH)₂D₃可加重气道重塑.

Abstract:

Objective To study the effects of 1,25-(OH)₂D₃ on airway remodeling and expression of high mobility group box 1 (HMGB1) and IL-17 in asthmatic mice. Methods Fifty female mice were randomly divided into 5 groups: control, asthma, low-dose, middle-dose, and high-dose intervention groups (n=10 each). Asthma was induced by intraperitoneal injections of ovalbumin (OVA) and aerosol inhalation of OVA solution. The low-dose, middle-dose, and high-dose intervention groups were administered with 1,25-(OH)₂D₃ solution at the dosage of 1, 4 and 10 μg/kg respectively by intraperitoneal injections before asthma challenge. The airway structural changes were assessed by hematoxylin and eosin staining. mRNA expression levels of HMGB1 and IL-17 in the lung tissues were evaluated by RT-PCR. The protein levels of HMGB1 and IL-17 in the lung tissues were observed by immunohistochemistry. Results The airway wall thickness, protein and mRNA expression levels of HMGB1 and IL-17 were higher in the untreated asthma group than in the control group (P<0.05). The airway wall thickness, protein and mRNA expression levels of HMGB1 and IL-17 were lower in the middle-dose and low-dose intervention groups than in the untreated asthma group, and the middle-dose intervention group demonstrated lower airway wall thickness, protein and mRNA expression levels of HMGB1 and IL-17 than in the low-dose intervention group (P<0.05). However, the airway wall thickness, protein and mRNA expression levels of HMGB1 and IL-17 in the high-dose intervention group were higher than in the untreated asthma group (P<0.05). Conclusions HMGB1 and IL-17 may be involved in the airway remodeling process in asthmatic mice. A moderate amount of HMGB1 and IL-17 may be involved in the airway remodeling process in asthmatic mice. A moderate amount of 1,25-(OH)₂D₃ can improve the airway remodeling, but a higher dose of 1,25-(OH)₂D₃ may affect adversely the airway remodeling process.