泼尼松对支气管上皮细胞黏蛋白5AC、细胞间黏附分子-1、白细胞介素-6分泌的影响

目的:探讨泼尼松对脂多糖(LPS)诱导的支气管上皮细胞黏蛋白5AC(MUC5AC)、细胞间黏附分子-1(ICAM-1)、白细胞介素-6(IL-6)分泌的影响。方法:体外分离鉴定原代支气管上皮细胞,LPS以1μg/m L剂量诱导细胞炎症反应,泼尼松5、10、20、40、50、100μmol/L干预治疗,酶联免疫法(ELISA)检测细胞上清液MUC5AC、ICAM-1、IL-6水平变化,采用定量逆转录PCR(RT-PCR)、免疫印迹(Western-blot)检测MUC5AC、ICAM-1、IL-6 m RNA和蛋白表达情况,免疫荧光法检测核因子κB(NF-κB)核定位,Western-blot检测泼尼松干预治疗前后表皮生长因子受体(EGFR)、细胞外调节蛋白激酶1/2(ERK1/2)活化情况。结果:泼尼松以剂量依赖性降低LPS诱导的支气管上皮细胞内MUC5AC、ICAM-1、IL-6 m RNA水平和蛋白水平,抑制NF-κB、EGFR、ERK1/2活化。结论:泼尼松在体外能抑制MUC5AC、ICAM-1、IL-6的产生,其机制可能与抑制NF-κB、EGFR、ERK1/2的激活有关。

Effects of Prednisone on Secretion of Mucin-5 Subtype AC, Ntercellular Adhesion Molecule-1 and Interleukin-6 in Bronchial Epithelial Cells

Objective:To probe into the effects of prednisone on secretion of mucin-5 subtype AC(MUC5 AC),intercellular adhesion molecule-1(ICAM-1)and interleukin-6(IL-6)in lipopolysaccharide(LPS)induced human bronchial epithelial cells.Methods:Primary human bronchial epithelial cells were isolated and identified in vitro.And 1μg/m L of LPS was used to induce cell inflammatory response.And 5,10,20,40,50,100μmol/L of prednisone was used as intervention treatment.The levels of MUC5 AC,ICAM-1 and IL-6 in cell supernatants was measured by euzymelinked immunosorbent assay(ELISA).The expression levels of MUC5 AC,ICAM-1,IL-6 m RNA and protein were determined by real-time PCR(RT-PCR)and Western-blot.Immunofluorescence method was used to detect nuclear localization of nuclear factor-κB(NF-κB).Epidermal growth factor receptor(EGFR)and extracellular signal-regulated kinase1/2(ERK1/2)phosphorylation before and after prednisone intervention were determined by Western-blot.Results:Prednisone inhibited the expression levels of MUC5 AC,ICAM-1,IL-6 m RNA and protein in a dose-dependent manner in LPS induced human bronchial epithelial cells.Meanwhile,prednisone suppressed NF-κB,EGFR and ERK1/2 phosphorylation.Conclusion:In response to LPS induced inflammation,prednisone inhibits the expression levels of MUC5 AC,ICAM-1 and IL-6 in vitro,which may be related to NF-κB,EGFR and ERK1/2 activation.