波动的压力对兔角膜内皮细胞的影响  

Effects of fluctuated pressure on corneal endothelial cells of rabbits

作　　者：幸正茂 梁玲玲[2] 袁进[3] 

机构地区：[1]南昌爱尔眼科,江西省南昌市330006 [2]江西新视界眼科医院,南昌大学,江西省南昌市330006 [3]中山大学中山眼科中心,广东省广州市510060

出　　处：《眼科新进展》2015年第12期1125-1128,共4页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:30801263)~~

摘　　要：目的观察波动的压力对角膜内皮细胞的影响。方法将体外培养的第一代兔角膜内皮细胞分为两组:A组为压力波动组(压力设置为:15 mm Hg^25 mm Hg^20 mm Hg^10 mm Hg,每个压力持续6 h;1 kPa=7.5 mm Hg);B组为30 mm Hg压力组;C组为无压力组。三组细胞分别培养24 h。免疫细胞化学染色法鉴定原代角膜内皮细胞形态;台盼蓝-茜素红联合染色检测细胞活性,HE染色观察细胞形态;Western-blotting检测细胞中Bcl-2和P53蛋白的表达水平。结果获取的所有细胞证实为角膜内皮细胞表型,无角膜上皮细胞及基质细胞污染。三组细胞分别培养24 h后,经台盼蓝-茜素红染色和HE染色证实:两个压力培养组的细胞活性较无压力培养组明显下降,其中压力波动组的细胞活性低于30 mm Hg压力组。同时无压力组、30 mm Hg压力组和压力波动组细胞中P53蛋白的相对表达量分别为0.150±0.005、0.253±0.014、0.670±0.019,差异有统计学意义(P<0.05)。结论证实了高压力及非生理性波动的压力对角膜内皮细胞均具有损伤作用。Objective To investigate the effects of fluctuated pressure bionic culture on rabbit corneal endothelial cells.Methods The rabbit corneal endothelial cells were divided into three groups,Group A：Fluctuated pressures were 15 mmHg-25 mmHg-20 mmHg- 10 mmHg（1 kPa = 7.5 mmHg）,each pressure was for 6 hours;Group B：The cells were exposed to 30 mmHg pressure in vitro;Group C：Without pressure culture group,as a blank control group,the three groups were cultured for 24 hours,respectively.Cells origin were identified by neuronalspecific enolase immunoassay,the cellular change in the structure were observed by HE staining,the cell activity was detected by trypan blue-alizarin red staining,and the expression level of Bcl-2 and P53 protein were detected by Western-blotting.Results NSE antibody of the primary corneal endothelial cells were positive without corneal epithelial cells and corneal stromal cells.Cells were cultured for 24 hours respectively,cell activity in two pressure groups was more decreased than that of group C by HE staining and blue staining with trypan blue-alizarin red staining,the cell activity of the fluctuated group was lower than that of 30 mmHg group.In the no pressure group,pressure fluctuated group and 30 mmHg group,the relative amount of P53 protein expression were 0.150 ±0.005,0.253 ±0.014,0.670 ±0.019,respectively,the relative amount of P53 protein expression was statistically significant（P〈0.05）.Conclusion The experiment confirm that high pressure and abnormal fluctuated pressure will cause damage to corneal endothelial cells.

关 键 词：压力 仿生培养 角膜内皮细胞 微环境 

分 类 号：R772.2[医药卫生—眼科]