中性粒细胞哮喘与嗜酸性粒细胞哮喘小鼠中Kv1.3钾离子通道表达及电流改变

目的 Kv1.3钾离子通道在与T细胞介导相关疾病的发生有一定的联系.该研究旨在研究Kv1.3钾离子通道在中性粒细胞及嗜酸性粒细胞哮喘模型中的表达及功能变化.方法 选择24只Balb/c小鼠,随机分成3组:正常对照组、中性粒细胞性哮喘组、嗜酸性粒细胞哮喘组.应用卵清蛋白联合内毒素建立哮喘模型,无创肺功能仪测定小鼠气道反应性、肺组织HE染色观察肺部炎症改变、免疫组织荧光检测Kv1.3钾离子通道在肺组织的定位、Western blot检测肺组织中Kv1.3钾离子通道蛋白表达量、全细胞膜片钳技术Kv1.3钾离子通道电流强度.结果 Kv1.3钾离子通道在哮喘气道及肺泡上皮区域表达增高,在中性粒细胞哮喘模型中表达强于嗜酸性粒细胞哮喘模型.中性粒细胞哮喘模型及嗜酸性粒细胞哮喘模型中肺泡灌洗液淋巴细胞上Kv1.3电流强度增加,且中性粒细胞哮喘模型增幅强于嗜酸性粒细胞哮喘模型.结论 Kv1.3钾离子通道在哮喘气道及肺泡上皮区域表达增高,电流强度增高.且在中性粒细胞哮喘模型中表达强于嗜酸细胞哮喘模型,为哮喘的治疗提供新的靶点.

Kv1.3 channel current changes in neutrophilic asthma and eosinophilic asthma

Objective Allergic asthma is a chronic inflammatory disease of the airways.T lymphocytes play important roles in the pathogenesis of asthma.The voltage-gated Kvl.3 potassium channel may be a key factor in the activation of T lymphocytes.This research aims to detect the function of Kvl.3 channel in the neutrophlial asthma(NA) model and eosinophilic asthma(EA) model.Methods A total of 24 mice were randomly assigned into three groups:control,neutrophilic asthma model and eosinophilic asthma group.Neutrophilic asthma model was established with ovalbumin (OVA)and lipopolysaccharide(LPS);eosinophilic asthma was established with OVA and Al(OH)3;airway responsiveness of mice in each group was measured with a noninvasive pulmonary function instrument;lung inflammation changes were observed by pathological HE staining;IL-17A and IL-4 cytokines levels in bronchoalveolar lavage fluid were evaluated by ELISA;Kvl.3 channel protein level in lung was evaluated by western blot;the change of current density in CD4 +.T lymphocytes were tested by whole-cell patch clamp technique.Results Levels of IL-17A and IL-4 in bronchoalveolar lavage fluid increased in both NA and EA model (P ＜ 0.05).Compare with EA model,the IL-17A level was significantly higer in NA model,while the IL-4 level was significantly lower.In NA and EA model,kv1.3 protein expression in lung tissue was significantly higher than that in the control group(P ＜ 0.05),and kv1.3 protein expression in NA model was significantly higer than that in the EA model (P ＜ 0.05).Current intensity and current density of kv1.3 channel increased in both NA and EA model.While the current intensity and current density of kv1.3 channel were significantly higher in NA model than that in EA model.Conclusion Kv1.3 protein level,Kv1.3 channel current intensity and kv1.3 channel current density increased in both NA and EA model,especially in NA group,providing a new way for treatment of bronchial asthma.