豚鼠内耳微血管内皮细胞的跨细胞电阻  

Transendothelial electrical resistance of microvascular endothelial cells from inner ear of guinea pigs

作　　者：魏运军[1] 张学渊[1] 

机构地区：[1]第三军医大学附属西南医院耳鼻咽喉科,重庆400038

出　　处：《第三军医大学学报》2002年第4期418-420,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目 ( 39870 834 )

摘　　要：目的　研究豚鼠内耳微血管内皮细胞跨细胞电阻的动态变化及峰值大小。方法　①胶原酶消化法分离培养豚鼠内耳及脑微血管内皮细胞 ,免疫组化法进行鉴定 ;②建立起内皮细胞通透性的体外模型 ,研究该模型下内耳内皮细胞的跨细胞电阻 ,并与脑及肺内皮细胞进行比较。结果　①培养细胞致密融合时具有内皮细胞培养时典型的“铺路石样”外观 ,与肺内皮细胞株的形态一致 ,经免疫组化检测其内皮细胞标志性抗原Ⅷ因子 ,95 %以上的培养细胞的胞浆中呈棕黄色阳性反应 ;②三种内皮细胞在培养增殖过程中其跨细胞电阻值呈动态变化过程 ,以 5× 10 4 cm2 的细胞密度接种时 ,7d左右电阻到达峰值 ,内耳、脑及肺三种内皮细胞的跨细胞电阻峰值大小分别为 (118 9± 18 5 )Ω cm2 ,(2 44 7± 46 9)Ω cm2 ,(4 6 6± 5 9)Ω cm( n =6) ,三者相差非常显著 (P <0 0 0 1)。结论　内耳微血管内皮细胞跨细胞电阻峰值小于脑内皮而大于肺内皮细胞 ,证明了三者通透性的差别。Objective To study the dynamic changes and the peak of transendothelial electrical resistance (TEER) of inner ear microvascular endothelial cells (EMEC) of guinea pigs. Methods ① EMEC and brain microvascular endothelial cells (BMEC) were isolated from guinea pigs and cultured with collagenase digestion. The cells were identified with immunohistochemical methods; ② After the establishment of the model for determining the permeability of endothelial cells in vitro , the TEER of EMEC was measured on it, and the results were compared with those of BMEC and lung microvascular endothelial cells (LMEC). Results ①Consistent with the features of LMEC, the monolayer confluent cells we cultured showed typical 'flagstone'appearance as cultured endothelial cells. Immunohistochemical detection of factor VIII, a mark antigen of endothelial cells, verified that over 95% of cultivated cells were positive to the factor with cytoplasmic buffy staining ② The TEERs of the 3 endothelial cells changed dynamically and reached the peak about 7 d after the cultured cells were inoculated at a 5×10 4/cm 2 dosage. The TEER's peak values of EMEC, BMEC and LMEC were 118.9±18.5, 244. 7±46.9, and 46.6±5.9Ω/cm 2 respectively ( n =6). Very significant differences were seen among the 3 peaks ( P <0.001). Conclusion The TEER peak of EMEC is lower than that of BMEC but greater than that of LMEC, which indicating the difference on their permeability.

关 键 词：豚鼠 内耳 微血管 内皮细胞 通透性 跨内皮细胞电阻 

分 类 号：R764.3[医药卫生—耳鼻咽喉科] R339.16[医药卫生—临床医学]