白细胞介素-13对人支气管上皮细胞SPDEF表达的影响及SPDEF在哮喘气道黏液高分泌中的作用

目的探讨白细胞介素(IL)-13对人支气管上皮细胞SPDEF表达的影响及SPDEF在哮喘气道黏液高分泌中的作用。方法将人原代支气管上皮细胞(NHBE细胞)分为对照组、IL-13组及IL-13+SPDEF siRNA组。培养28天后,收集NHBE细胞并提取总RNA,采用实时荧光定量聚合酶链反应检测NHBE细胞SPDEF、粘蛋白MUC5AC及MUC5B mRNA的表达水平,流式细胞术检测MUC5AC阳性细胞数量和免疫荧光强度,免疫荧光染色检测粘蛋白MUC5AC和MUC5B的表达水平。结果IL-13组NHBE细胞SPDEF和MUC5AC mRNA的表达水平、MUC5AC阳性细胞数量和免疫荧光强度均明显高于对照组(P<0.001),而MUC5B mRNA的表达水平明显低于对照组(P<0.05);IL-13+SPDEF siRNA组NHBE细胞SPDEF和MUC5AC mRNA的表达水平、MUC5AC阳性细胞数量和免疫荧光强度均明显低于IL-13组(P<0.001);IL-13+SPDEF siRNA组MUC5B mRNA的表达水平明显低于对照组(P<0.05)。IL-13组NHBE细胞MUC5AC的表达水平明显高于对照组和IL-13+SPDEF siRNA组,而MUC5B的表达水平低于对照组。结论IL-13可能通过诱导人支气管上皮细胞SPDEF上调,促进气道粘蛋白MUC5AC的高表达,引起哮喘气道黏液高分泌。

Effects of interleukin-13 on SPDEF expression in human bronchial epithelial cells and role of SPDEF in airway mucus hypersecretion of asthma

Objective To explore the effects of interleukin(IL)-13 on SPDEF expression in human bronchial epithelial cells and role of SPDEF in airway mucus hypersecretion of asthma.Methods Normal human bronchial epithelial(NHBE)cells were divided into control group,IL-13 group and IL-13+SPDEF siRNA group.After 28 days of the culture,NHBE cells were collected and total RNA was extracted.Real-time polymerase chain reaction was used to detect the expression levels of SPDEF,MUC5AC and MUC5B mRNA in NHBE cells.Numbers and immunofluorescence intensity of MUC5AC positive cells were identified by flow cytometry.Eexpression levels of mucin MUC5AC and MUC5B were detected by immunefluorescence staining.Results Expression levels of SPDEF and MUC5AC mRNA,numbers and immunofluorescence intensity of MUC5AC positive cells in IL-13 group were significantly higher than those in control group(P<0.001),while expression level of MUC5B mRNA was significantly lower than that in control group(P<0.05).The expression levels of SPDEF and MUC5AC mRNA,numbers and immunofluorescence intensity of MUC5AC positive cells in IL-13+SPDEF siRNA group were significantly lower than those in IL-13 group(P<0.001).Expression level of MUC5B mRNA in IL-13+SPDEF siRNA group was lower than that in control group(P<0.05).The expression level of MUC5AC in NHBE cells in IL-13 group was significantly higher than those in control group and IL-13+SPDEF siRNA group,while the expression level of MUC5B was lower than that in control group.Conclusion IL-13 may promote the hyperexpression of mucin MUC5AC by inducing the up-regulation of SPDEF in human bronchial epithelial cells,leading to airway mucus hypersecretion of asthma.