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[紧密连接蛋白闭合蛋白在鼻息肉中的表达及调控]

[Expression and regulation of tight junction protein Occludin in nasal polyps].

作者信息

Chen Xi, Guo Jiebo, Zhang Han, Zuo Kejun, Wei Yi, Shi Jianbo, Li Huabin, Xu Geng

机构信息

Department of Otorhinolaryngology Head and Neck Surgery, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, China.

Department of Otorhinolaryngology Head and Neck Surgery, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, China. Email:

出版信息

Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2014 Jul;49(7):568-73.

Abstract

OBJECTIVE

To evaluate the possible role of tight junction protein Occludin in nasal polyps.

METHODS

The expression of Claudin-1, Occludin and ZO-1 in nasal polyps (n = 20) and healthy uncinate mucosa (n = 15) were examined using immunohistochemical staining, real-time quantitative polymerase chain reaction (PCR) and Western blot analysis. The regulatory effects of proinflammatory cytokines (IFN-γ, IL-13, IL-17, TGF-β, TGF-α) on the expression of Occludin in cultured human nasal epithelial cells were investigated.

RESULTS

The immunohistochemical results showed that Claudin-1, Occludin and ZO-1 were detected both in the nasal polyp group and the control group. The expression sites were the cell membrane and cytoplasm of nasal mucosa epithelial cells. The mean optical density of Claudin-1, Occludin and ZO-1 were 0.187 ± 0.076,0.172 ± 0.109 and 0.098 ± 0.035 respectively in the nasal polyp group and were significantly lower than those in the control group (0.312 ± 0.101, 0.220 ± 0.069 and 0.233 ± 0.093 respectively), the differences were significant (t = 9.345, t = 3.301, t = 13.323, all P < 0.01).RT-PCR results showed that the relative expression of Occludin mRNA was 0.000 117 ± 0.000 035 in the nasal polyp group and was significantly lower than that in the control group(0.000 464 ± 0.000 134), and the difference was significant (Z = -5.0, P < 0.01) . There was no statistically significant difference in the relative expression of Claudin-1 and ZO-1 mRNA between the nasal polyp group and the control group (P > 0.05) . After the cultured human nasal epithelial cells were stimulated by IL-13, IL-17, IFN-γ and other proinflammatory cytokines, the relative expression of Occludin mRNA was 0.631 ± 0.039, 0.581 ± 0.029 and 0.648 ± 0.040, respectively. Compared with the unstimulated control group, the differences were statistically significant (t = 16.299, 24.669 and 14.995 respectively, all P < 0.05).Western blot analyse showed that the relative grayscale in the above proinflammatory cytokines stimulation groups was 0.650 ± 0.061,0.482 ± 0.106 and 0.536 ± 0.109, respectively. Compared with the unstimulated control group, the differences were statistically significant (t = 9.880, 8.442 and 7.310 respectively, all P < 0.05).

CONCLUSIONS

The reduced expression of Occludin might be involved in the pathogenesis of nasal polyps.

摘要

目的

评估紧密连接蛋白闭合蛋白(Occludin)在鼻息肉中的可能作用。

方法

采用免疫组织化学染色、实时定量聚合酶链反应(PCR)及蛋白质印迹分析,检测20例鼻息肉组织及15例健康钩突黏膜中Claudin-1、Occludin和闭锁小带蛋白1(ZO-1)的表达。研究促炎细胞因子(γ干扰素、白细胞介素13、白细胞介素17、转化生长因子β、转化生长因子α)对培养的人鼻上皮细胞中Occludin表达的调控作用。

结果

免疫组织化学结果显示,鼻息肉组和对照组均检测到Claudin-1、Occludin和ZO-1。表达部位为鼻黏膜上皮细胞的细胞膜和细胞质。鼻息肉组Claudin-1、Occludin和ZO-1的平均光密度分别为0.187±0.076、0.172±0.109和0.098±0.035,显著低于对照组(分别为0.312±0.101、0.220±0.069和0.233±0.093),差异有统计学意义(t=9.345、t=3.301、t=13.323,P均<0.01)。实时定量PCR结果显示,鼻息肉组Occludin mRNA的相对表达量为0.000 117±0.000 035,显著低于对照组(0.000 464±0.000 134),差异有统计学意义(Z=-5.0,P<0.01)。鼻息肉组与对照组Claudin-1和ZO-1 mRNA的相对表达量差异无统计学意义(P>0.05)。人鼻上皮细胞经白细胞介素13、白细胞介素17、γ干扰素等促炎细胞因子刺激后,Occludin mRNA的相对表达量分别为0.631±0.039、0.581±0.029和0.648±0.040。与未刺激的对照组相比,差异有统计学意义(t分别为16.299、24.669和14.995,P均<0.05)。蛋白质印迹分析显示,上述促炎细胞因子刺激组的相对灰度值分别为0.650±0.061、0.482±0.106和0.536±0.109。与未刺激的对照组相比,差异有统计学意义(t分别为9.880、8.442和7.310,P均<0.05)。

结论

Occludin表达降低可能参与鼻息肉的发病机制。

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