Role of p38/MAPK pathway in hyperosmosis-induced functional impairment of human corneal epithelial barrier <italic style="font-style: italic">in vitro</italic>

Shuang QIN; Wenjun ZHOU; Hua LI; Shengfang SONG

doi:10.16016/j.1000-5404.201707055

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Role of p38/MAPK pathway in hyperosmosis-induced functional impairment of human corneal epithelial barrier in vitro

Basic Medicine | 更新时间：2026-01-09

- Role of p38/MAPK pathway in hyperosmosis-induced functional impairment of human corneal epithelial barrier in vitro
- Journal of Third Military Medical University Vol. 39, Issue 23, Pages: 2282-2288(2017)
- 作者机构：
  
  402160 重庆，重庆医科大学附属永川医院眼科
- 作者简介：
- 基金信息：
  
  the Basic Science and Frontier Technology Research Project of Chongqing(CSTC2016jcyjA0249);the Post-graduate Innovation Fund of Yongchuan Hospital of Chongqing Medical University(YJSCX201602)
- DOI：10.16016/j.1000-5404.201707055
  CLC： R322.91;R329.24;R363.22
- Received：06 July 2017，
  
  Revised：2017-09-09，
  
  Published：15 December 2017
- 稿件说明：
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Shuang QIN, Wenjun ZHOU, Hua LI, et al. Role of p38/MAPK pathway in hyperosmosis-induced functional impairment of human corneal epithelial barrier in vitro[J]. Journal of Third Military Medical University, 2017, 39(23): 2282-2288.
DOI：

Shuang QIN, Wenjun ZHOU, Hua LI, et al. Role of p38/MAPK pathway in hyperosmosis-induced functional impairment of human corneal epithelial barrier in vitro[J]. Journal of Third Military Medical University, 2017, 39(23): 2282-2288. DOI： 10.16016/j.1000-5404.201707055.

摘要

目的

探讨高渗透压对角膜上皮屏障功能的影响及p38/MAPK信号通路在此过程中潜在的作用。

方法

体外培养人永生化角膜上皮细胞(human corneal epithelial cells，HCEC)，用不同浓度渗透压[302(正常渗透压)、350、500 mOsm/L]作用于HCEC细胞24 h，CCK-8法检测细胞增殖活性；免疫荧光检测细胞紧密连接蛋白ZO-1和Claudin-1的表达和分布；p38/MAPK阻断剂预处理HCEC细胞前后，Western blot分别检测紧密连接蛋白ZO-1、Claudin-1和p38/MAPK信号通路表达及其磷酸化水平；跨膜电阻仪检测上皮细胞跨膜电阻(transepithelial electrical resistance

TEER)的变化。

结果

高渗透压对HCEC细胞增殖活性没有影响；免疫荧光染色可见正常渗透压组ZO-1、Claudin-1沿着细胞膜呈线性分布，而350、500 mOsm/L高渗透压组ZO-1、Claudin-1荧光信号均减弱，且细胞间紧密连接结构破坏(

0.05)；在高渗透压下，HCEC细胞紧密连接蛋白ZO-1、Claudin-1表达量明显下降(

0.05)，p38/MAPK磷酸化水平增加(

0.05)，上皮细胞跨膜电阻(TEER)降低(

0.05)，而p38/MAPK阻断剂SB203580可抑制高渗透压引起的上述反应(

0.05)。

结论

高渗透压可以使人角膜上皮细胞屏障功能破坏，其机制为通过激活p38/MAPK信号通路导致紧密连接蛋白ZO-1、Claudin-1表达下降。

Abstract

Objective

To study the effect of hyperosmosis on corneal epithelial barrier function and explore the role of p38/MAPK pathway in this process.

Methods

Human corneal epithelial cells (HCECs) cultured in normal osmotic (302 mOsm/L) or in hyperosmotic (350 and 500 mOsm/L) conditions for 24 h were examined for cell proliferation using CCK-8 assay and for distribution of tight junction proteins zonula occludens 1 (ZO-1) and claudin-1 using immunofluorescence analysis. Western blotting was used to detect the expression of ZO-1

claudin-1

p38 and phosphorylation of p38 in the cells cultured in different osmotic conditions with or without prior treatment of 10 μg/L SP203580

a p38/MAPK specific inhibitor. The transepithelial electrical resistance (TEER) of the cells was measured using a transmembrane resistance meter.

Results

Hyperosmosis produced no obvious effect on HCEC cell proliferation. Immunofluorescence assay showed that in the cells cultured in normal osmotic condition

ZO-1 and claudin-1 emitted fluorescence in a linear pattern along the cell membrane

while the cells cultured at 350 or 500 mOsm/L exhibited a significantly lowered intensity of the fluorescence signals with obvious structural collapse of the tight junctions (

0.05). In hyperosmotic conditions

the cells expressed significantly decreased levels of tight junction proteins ZO-1 and claudin-1 (

0.05) with significantly increased phosphorylation of p38 (

0.05) and reduced TEER (

0.05). Treatment of the cells with the p38 inhibitor (SB203580) prior to hyperosmotic exposures obviously suppressed these responses to hyperosmosis (

0.05).

Conclusions

Hyperosmosis induces impairment of corneal epithelial barrier function by activating the p38/MAPK pathway to cause down-regulation of the tight junction proteins ZO-1 and claudin-1.

关键词

Keywords

references

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Role of p38/MAPK pathway in hyperosmosis-induced functional impairment of human corneal epithelial barrier in vitro

DOI：10.16016/j.1000-5404.201707055

摘要

Abstract

关键词

Keywords

references