[肿瘤坏死因子-α对豚鼠耳蜗血管纹毛细血管内皮细胞通透性的影响]

[Effect of tumor necrosis factor-alpha on the permeability of strial capillary endothelial cells in guinea pig cochlea].

作者信息

Li Qi, Yuan Wei, Wei Yun-Jun, Zhang Xue-Yuan

机构信息

Department of Otorhinolarynogology Head and Neck Surgery, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.

出版信息

Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2008 Sep;43(9):691-5.

PMID:19035264

Abstract

OBJECTIVE

To perform an initial study on possible molecular regulatory mechanisms of TNF-alpha to the permeability of strial capillary endothelial cells in guinea pig cochlea

METHODS

Strial capillary endothelial cells in guinea pig cochlea was dissociated and cultured to establish a model for its permeability in vitro. The animals were divided into TNF-alpha, TNF-alpha + L-arginine (L-Arg), TNF-alpha + NG-monomethyl-L-arginine (L-NMMA) and control groups. In order to measure the alternations of permeation rate, Evans blue was used in different time by different concentrations to detect contents of F-actin in each group under immunofluorescence laser scanning confocal microscope.

RESULTS

TNF-alpha increased permeability of strial capillary endothelial cells notably, and permeation rate to Evans blue heightened significantly (P < 0.01) and increased under higher concentrations of TNF-alpha as well as lengthened the durations. Being as a NO donator, L-Arg significantly enhanced the ability of TNF-alpha which increasing permeability of strial capillary endothelial cells; permeation rate to Evans blue heightened significantly (P < 0.01), which obviously rose up when the concentration of Evans blue added. L-NMMA, NO inhibitor, was able to weaken the ability of TNF-alpha which increasing permeability of strial capillary endothelial cells; permeation rate to Evans blue decreased significantly (P < 0.01), which obviously fell down when the concentration of L-NMMA added. TNF-alpha decreased F-actin in strial capillary endothelial cells (P < 0.01), more TNF-alpha, less F-actin; L-Arg could be further the inhibition to F-actin content (P < 0.05); L-NMMA held back the trend (P < 0.05).

CONCLUSIONS

TNF-alpha can increase permeability of strial capillary endothelial cells of guinea pig cochlea. NO may be one of the important molecular regulators to permeability of strial capillary endothelial cells by TNF-alpha, and changes of F-actin content probably relate to the regulation in these cells.

摘要

目的

对肿瘤坏死因子-α（TNF-α）影响豚鼠耳蜗血管纹毛细血管内皮细胞通透性的可能分子调控机制进行初步研究。

方法

分离培养豚鼠耳蜗血管纹毛细血管内皮细胞，建立体外通透性模型。将动物分为TNF-α组、TNF-α+L-精氨酸（L-Arg）组、TNF-α+N-甲基-L-精氨酸（L-NMMA）组和对照组。为检测渗透率的变化，在不同时间使用不同浓度的伊文思蓝，通过免疫荧光激光扫描共聚焦显微镜检测各组中F-肌动蛋白的含量。

结果

TNF-α显著增加血管纹毛细血管内皮细胞的通透性，伊文思蓝渗透率显著升高（P<0.01），且在较高浓度的TNF-α作用下增加，并延长了作用时间。作为一氧化氮（NO）供体，L-Arg显著增强了TNF-α增加血管纹毛细血管内皮细胞通透性的能力；伊文思蓝渗透率显著升高（P<0.01），添加伊文思蓝时明显上升。NO抑制剂L-NMMA能够削弱TNF-α增加血管纹毛细血管内皮细胞通透性的能力；伊文思蓝渗透率显著降低（P<0.01），添加L-NMMA时明显下降。TNF-α降低血管纹毛细血管内皮细胞中的F-肌动蛋白（P<0.01），TNF-α越多，F-肌动蛋白越少；L-Arg可进一步抑制F-肌动蛋白含量（P<0.05）；L-NMMA抑制了这一趋势（P<0.05）。