Objective: To check Forkhead box M1 (FoxM1) expression in nasal mucosal of chronic rhinosinusitis (CRS) patients and the effect of inflammatory factors on FoxM1 expression, in order to research the significance of FoxM1 in CRS. Methods: From January to October of 2018, 50 patients hospitalized in the Department of Otorhinolaryngology, Head and Neck Surgery, First Affiliated Hospital of Nanchang University were enrolled in this study. Twenty CRS patients with polyps (CRSwNP), 20 CRS patients without nasal polyps (CRSsNP) and 10 patients with simple deviation of nasal septum (the control groups) were selected. The expression of FoxM1 in nasal mucosa of these patients was detected by immunohistochemistry (IHC) and real-time fluorescent quantitative PCR (qRT-PCR). Meanwhile, HE stain was used to observe the pathologic changes in each sample. By establishing human nasal epithelium cells cultivating model in vitro and identifying via immumofluorescence method, experimental group and control group were set up, then activation factors including interleukin (IL)-1β, IL-4, IL-5, IL-17, interferon-γ (IFN-γ) and staphylococcal entemtoxin B (SEB) were added in the models after stabilizing passage, and qRT-PRC and Western blot method were applied to check the expressing change of FoxM1. Software SPSS 18.0 was used for statistical analysis. Results: HE stain showed that the mainly pathologic change in nasal mucosa of CRS patients with or without nasal polyp was mucosal epithelial cells, goblet cell and submucosal gland hyperplasia, accompanied by a large number of inflammatory cells infiltration. The result of IHC demonstrated that both of the expression of FoxM1 in nasal mucosal tissue of CRS patients in the CRSwNP and CRSsNP groups exceed that of the control group (80% vs 75% vs 20%, χ(2) value was 10.000, 8.213, respectively, all P<0.05); there was no difference of expression between the two groups of CRS patients (χ(2)=0.143, P>0.05). The result of qRT-PCR demonstrated that the expression of FoxM1 mRNA in nasal mucosa of CRSwNP and CRSsNP was increased compared with that of the control group (3.309±1.511 vs 3.261±1.336 vs 1.000±0.774, t value was 4.519, 4.928, respectively, all P<0.05), but the difference between the two groups of CRS patients had no statistic significance (t=0.107, P=0.909). Nasal mucosa epithelial cells cultivating models was established successfully. Q-RT PCR and Western blot were conducted after stimulation of 100 ng/ml IL-1β, IL-4, IL-5, IL-17, IFN-γ and SEB for 36 h, and the proteins expression levels of FoxM1 exceeded the groups without stimulation with statistic significance. Conclusions: The expression of FoxM1 in CRS increases and many types of cytokine can induce the increase of FoxM1 in human nasal epithelial cells. FoxM1 may participate in the process of pathogenesis in CRS.
目的： 检测转录因子叉头框M1（Forkhead box M1，FoxM1）在慢性鼻窦炎（chronic rhinosinusitis，CRS）患者鼻黏膜中的表达情况及炎性因子对鼻黏膜细胞FoxM1表达的影响，以了解FoxM1在CRS发病中的作用。 方法： 2018年1—10月期间，于南昌大学第一附属医院耳鼻咽喉头颈外科住院的50例患者纳入本研究，其中伴鼻息肉CRS患者20例、不伴鼻息肉CRS患者20例、单纯鼻中隔偏曲患者10例。取鼻黏膜组织行苏木精-伊红（HE）染色，查看组织病理特点，并行免疫组织化学（immunohistochemistry，IHC）、实时荧光定量PCR（qRT-PCR）检测组织中FoxM1的水平。体外培养人鼻黏膜上皮细胞并进行鉴定，设立实验组及空白对照组。实验组分别给予炎性因子白细胞介素（interleukin，IL）1β、IL-4、IL-5、IL-17、γ干扰素（interferon-γ，IFN-γ）、葡萄球菌肠毒素B（staphylococcal entemtoxin B，SEB）后，应用qRT-PCR法、免疫印记法（Western blot法）检测实验组及空白对照组FoxM1的表达情况。应用SPSS 18.0对实验数据进行统计学分析。 结果： HE染色示CRS患者鼻黏膜上皮细胞及黏膜下腺体增生，黏膜下组织内可见大量炎性细胞浸润。IHC显示伴及不伴鼻息肉的CRS患者鼻黏膜组织中FoxM1的表达均较鼻中隔偏曲组升高（80%比75%比20%，χ(2)值分别为10.000、8.213，P值均<0.05），两组CRS患者之间FoxM1的表达差异无统计学意义（χ(2)=0.143，P>0.05）。qRT-PCR结果显示伴及不伴鼻息肉的CRS患者鼻黏膜组织中FoxM1 mRNA的表达均较鼻中隔偏曲组上调（3.309±1.511比3.261±1.336比1.000±0.774，t值分别为4.519、4.928，P值均<0.05），而两组CRS之间差异无统计学意义（t=0.107，P=0.909）。在体外进行了鼻黏膜原代细胞培养及鉴定，对稳定传代的鼻黏膜上皮细胞给予浓度均为100 ng/ml的IL-1β、IL-4、IL-5、IL-17、IFN-γ、SEB刺激36 h后，FoxM1的蛋白表达水平均较空白对照组上调，且差异有统计学意义。 结论： CRS患者鼻黏膜中FoxM1表达上调，且多种炎性因子可导致鼻黏膜上皮细胞FoxM1表达增多，提示FoxM1可能参与了CRS的发病过程。.
Keywords: Epithelial cells; Forkhead box M1; Interleukins; Nasal mucosa; Sinusitis.