布地奈德通过调节IL-4/IL-13/STAT6影响大鼠黏膜重塑

Budesonide suppresses nasal mucosal remodeling by regulating IL-4/IL-13/STAT6 pathway in rats with allergic rhinitis

目的探讨布地奈德通过IL-4/IL-13/STAT6信号通路对变应性鼻炎(allergic rhinitis,AR)大鼠鼻黏膜重塑的影响。方法 102只大鼠采用随机数字表法分为3组:空白组、AR-PBS组、AR-布地奈德组,每组34只。AR-PBS组及AR-布地奈德组大鼠为建模组大鼠,均采用卵清蛋白致敏的方法构建AR模型。建模成功后,使用微量移液枪双侧滴鼻治疗。AR-PBS组给予(PBS)治疗,AR-布地奈德组给予布地奈德(倍受您)治疗,空白组大鼠给予等量PBS治疗,治疗时间持续7 d。采用动物评价量表对各组大鼠行为学进行评分;ELISA法测定血清组胺(His)及Ig E;液相芯片法测定血清及肺泡灌洗液IL-4、IL-13浓度;定量PCR检测STAT6基因表达,Western blot检测鼻黏膜总STAT6及磷酸化p-STAT6蛋白表达;鼻中隔黏膜HE染色及透射电镜扫描观察鼻黏膜重塑情况;采用SPSS 23.0统计软件对数据进行分析处理。结果建模组大鼠建模结束时动物行为学评分均>5分,血清His、Ig E,血清及肺泡灌洗液IL-4及IL-13浓度以及鼻中隔黏膜STAT6及p-STAT6表达均显著高于空白组(P<0.01)。建模组大鼠采用不同方法治疗,AR-布地奈德组干预7 d后,血清His、Ig E,血清及肺泡灌洗液IL-4、IL-13浓度,以及鼻中隔黏膜STAT6、p-STAT6表达均下调,显著低于AR-PBS组,差异有统计学意义(P<0.01)。HE染色及透射电镜结果显示建模组大鼠建模结束时鼻纤毛减少,布地奈德治疗后纤毛具有一定程度的恢复,治疗前后细胞连接无显著差异。结论布地奈德通过降低IL-4、IL-13、p-STAT6从而改善AR大鼠鼻黏膜症状。

ObjectiveTo investigate the effect of budesonide on nasal mucosal remodeling and the role of IL-4/IL-13/STAT6 signaling pathway in mediating this effect in rats with allergic rhinitis （AR）. MethodsA total of 102 rats were randomly allocated into 3 equal groups, namely AR-PBS group, AR-budesonide group and blank group. The rats in AR-PBS and AR-budesonide groups were sensitized by challenges with ovalbumin to establish models of AR, and were subsequently treated by daily intranasal instillation with PBS and budesonide with aid of Eppendof for 7 consecutive days, respectively. The rats in the blank group were not challenged with ovalbumin and were treated with PBS in the same manner. Animal Assessment Scale was used to assess the behavioral changes of the rats after the treatments. Enzyme-linked immunosorbent assay （ELISA） was used to detect the serum levels of histamine （His） and IgE, and the levels of interleukin （IL）-13 and IL-4 in the serum and bronchoalveolar lavage were analyzed with a Luminex liquid chip. STAT6 mRNA expression in the nasal mucosal was assessed with qPCR, and the protein expression levels of STAT6 and P-STAT6 were tested with Western blotting. HE staining and transmission electron microscopy were used to assess nasal septum remodeling after the treatments. ResultsThe rat models of AR all had a behavioral assessment score over 5 by the Animal Assessment Scale. Compared with those in the blank group, the rats with AR showed significantly increased serum levels of His and IgE, increased levels of IL-13 and IL-4 in both the serum and bronchoalveolar lavage, and also increased expression of STAT6 and P-STAT6 in the nasal mucosa （P〈0.01）. Budesonide treatment of the rats with AR for 7 days obviously lowered the serum levels of His and IgE, which remained unchanged in rats with PBS treatment. Compared with PBS treatment, budesonide treatment also decreased the levels of IL-4 and IL-13 in the serum and bronchoalveolar lavage and down-regulated the expression of STAT6 and P-STAT6 in the nasal mucosa （P〈0.01）. HE staining and transmission electron microscopy showed that budesonide treatment partially reversed AR-induced reduction of nasal cilia in the rats without causing obvious changes in cell connection. ConclusionBudesonide improves the pathologies in the nasal mucosa of rats with AR by inhibiting IL-4, IL-13, and p-STAT6.