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目的探讨细胞外基质(extracellularmatrix,ECM)成分固生蛋白C(tenascinC,TNC)在鼻息肉组织中异常表达的原因。方法采用免疫组化方法检测TNC和转化生长因子β1(transforminggrowthfactor-β1,TGF-β1)在鼻息肉组织中的表达和关系。进一步采用细胞培养、实时定量RTPCR和原位ELISA技术研究TGFβ1及嗜酸粒细胞对人呼吸道上皮细胞系BEAS2B细胞TNC表达的调控作用。结果①TNC和TGF-β1蛋白在鼻息肉组织中的表达显著上调,TNC表达强度与TGFβ1阳性细胞总数(r=-0.58,P<0.01)和TGFβ1阳性的嗜酸粒细胞数显著相关(r=-0.61,P<0.01);②浓度为1ng/ml和10ng/ml的TGFβ1刺激4h后BEAS2B细胞TNCmRNA的表达分别为未刺激状态下的(7.20±3.43,x±s,下同)倍和(22.48±5.35)倍,与未刺激状态下的表达水平相比差异有统计学意义(P值<0.01);与刺激24h后TNC蛋白表达的荧光强度相比差异亦有统计学意义(P<0.05);③BEAS2B细胞和嗜酸粒细胞以2∶1、1∶1和1∶2的数量比例进行共培养,4h后BEAS2B细胞TNCmRNA的表达与未刺激状态下的表达水平相比,差异均有统计学意义(P值均<0.01);24h后TNC蛋白表达的荧光强度与未刺激状态下的荧光强度相比差异亦均有统计学意义(P值均<0.05);嗜酸粒细胞的这种诱导作用可以被抗TGFβ1的中和抗体显著抑制(P<0.05)。结论TGF-β1和嗜酸粒细胞可以诱导呼吸道上皮细胞对TNC的表达,嗜酸粒细胞的作用部分通过TGFβ1介导,鼻息肉组织中TNC表达的增高同嗜酸粒细胞来源的TGF-β1有关。
Objective To investigate the reason of abnormal expression of tenascinC (TNC), an extracellular matrix (ECM) component in nasal polyps. Methods The expressions of TNC and transforming growth factor β1 (TGF-β1) in nasal polyps were detected by immunohistochemistry. Furthermore, the effects of TGFβ1 and eosinophils on the TNC expression of human respiratory epithelial cell line BEAS2B were further studied by cell culture, real-time quantitative RTPCR and in situ ELISA. Results ① The expression of TNC and TGF-β1 protein in nasal polyps was significantly up-regulated. The expression of TNC was correlated with the number of TGFβ1 positive cells (r = -0.58, P <0.01) and the number of TGFβ1 positive eosinophils (r = (7.20 ± 3.43, x ± s, the same below) times and (22.48, 0.61, P <0.01) respectively; ②The expression of TNC mRNA in BEAS2B cells after treated with 1ng / ml and 10ng / ml TGFβ1 for 4h were ± 5.35) fold, which was significantly different from that of unstimulated (P <0.01). The difference was also statistically significant (P <0.05) compared with the fluorescence intensity of TNC after 24h stimulation ; ③BEAS2B cells and eosinophils were co-cultured in a ratio of 2: 1, 1: 1 and 1: 2. After 4h, the expression of TNC mRNA in BEAS2B cells was statistically different from that in unstimulated (P <0.01). After 24 h, the fluorescence intensity of TNC protein expression was also significantly different from that of unstimulated fluorescence intensity (P <0.05). This induction of eosinophils The effect can be significantly inhibited by anti-TGFβ1 neutralizing antibody (P <0.05). Conclusion TGF-β1 and eosinophils can induce the expression of TNC in airway epithelial cells. The eosinophil function is partially mediated by TGFβ1. The increased expression of TNC in nasal polyps is associated with eosinophil-derived TGF-β1 .