TET2表达下调与慢性鼻窦炎伴鼻息肉鼻黏膜上皮紧密连接损伤的相关性研究

A study on the correlation between downregulated TET2 expression and tight junction injury of nasal mucosal epithelium in chronic rhinosinusitis with nasal polyps

摘要:

目的 探讨甲基胞嘧啶双加氧酶2（TET2）在慢性鼻窦炎伴鼻息肉（CRSwNP）患者鼻黏膜上皮紧密连接调节中的作用。方法 收集2025年4月至12月武汉大学人民医院45例CRSwNP患者鼻息肉组织及20例健康对照者下鼻甲组织，分为嗜酸性CRSwNP组（ECRSwNP，24例）、非嗜酸性CRSwNP组（Non-ECRSwNP，21例）和对照组。采用qRT-PCR和免疫荧光染色检测各组TET2及紧密连接蛋白（ZO-1、Occludin、Claudin-3）的表达水平，分析紧密连接蛋白表达与Lund-Mackay评分的相关性；体外利用siRNA敲低人鼻黏膜上皮细胞（HNEpCs）中的TET2，通过跨上皮电阻和FITC-Dextran通透性实验评估屏障功能，并验证TET2对紧密连接蛋白表达的调控作用。结果 与对照组相比，CRSwNP患者鼻息肉组织中ZO-1及Claudin-3 mRNA及蛋白表达均显著下调（P<0.05），ECRSwNP组Occludin mRNA及蛋白表达显著下调，Non-ECRSwNP组Occludin mRNA下降无统计学意义，ECRSwNP与Non-ECRSwNP组上述指标比较差异无统计学意义（P>0.05）；ZO-1、Occludin及Claudin-3 mRNA表达水平与Lund-Mackay评分均呈负相关（r=-0.709、-0.572、-0.789，P<0.01）。TET2 mRNA表达与ZO-1、Occludin、Claudin-3 mRNA表达均呈显著正相关（r=0.524、0.533、0.679，P<0.01）。体外敲低TET2导致HNEpCs的跨上皮电阻进行性下降，FITC-Dextran通透性显著增加，并显著下调了ZO-1、Occludin和Claudin-3的mRNA及蛋白表达（P<0.05）。结论 TET2表达下调与紧密连接蛋白减少、上皮屏障损伤相关。TET2表达异常可能是CRSwNP上皮屏障功能受损的重要原因，为阐明CRSwNP的发病机制提供了新的视角。

Abstract:

Objective: To investigate the role of ten-eleven translocation 2 (TET2) in regulating tight junctions in nasal mucosal epithelium of patients with chronic rhinosinusitis with nasal polyps (CRSwNP). Methods: Nasal polyp tissues from 45 CRSwNP patients and inferior turbinate tissues from 20 healthy controls were collected at Renmin Hospital of Wuhan University between April and December 2025. Patients were divided into eosinophilic CRSwNP group (ECRSwNP, n=24), non-eosinophilic CRSwNP group (Non-ECRSwNP, n=21), and control group. Expression levels of TET2 and tight junction proteins (ZO-1, Occludin, Claudin-3) were analyzed by qRT-PCR and immunofluorescence staining. The correlation between tight junction protein expression and Lund-Mackay scores was evaluated. In vitro, TET2 was knocked down in human nasal epithelial cells (HNEpCs) using siRNA, and barrier function was assessed via transepithelial electrical resistance (TEER) and FITC-dextran permeability assays to validate TET2 regulatory effects on tight junction proteins. Results: Compared with the control group, the mRNA and protein expressions of ZO-1 and Claudin-3 in nasal polyp tissues of CRSwNP patients were significantly downregulated (P<0.05). The mRNA and protein expressions of Occludin were significantly downregulated in the ECRSwNP group, while the decrease of Occludin mRNA in the Non-ECRSwNP group was not statistically significant. There was no statistically significant difference in the above indicators between the ECRSwNP group and the Non-ECRSwNP group (P>0.05). The mRNA expression levels of ZO-1, Occludin and Claudin-3 were negatively correlated with the Lund-Mackay score (r=-0.709、-0.572、-0.789，P<0.01) . TET2 mRNA expression was significantly positively correlated with the mRNA expression of ZO-1, Occludin, and Claudin-3(r=0.524、0.533、0.679，P<0.01).In vitro knockdown of TET2 led to a progressive decline in TEER, a significant increase in FITC-dextran permeability, and marked downregulation of ZO-1, Occludin, and Claudin-3 at both mRNA and protein levels (P<0.05). Conclusion: Downregulation of TET2 expression is associated with reduced tight junction proteins and epithelial barrier injury.Aberrant TET2 expression may be a key factor underlying impaired epithelial barrier function in CRSwNP, offering new insights into the pathogenesis of this condition.