高温对体外培养的兔视网膜色素上皮细胞的影响

Effect of High Temperature on Cultured Rabbit Retinal Pigment Epithelium Cells

目的:观察高温对体外培养的兔视网膜色素上皮(retinal pigment epithelial cells,RPE)细胞的影响。方法:以体外培养的兔RPE细胞为研究对象,采用55℃水浴加热法对细胞进行不同时间(3、5、8min)加温处理,处理结束后立即进行CCK-8法检测细胞活性、流式细胞仪测定细胞凋亡、透射电镜观察细胞形态学变化。结果:55℃水浴各时间组OD值明显低于对照组,差异有统计学意义(P<0.05),且55℃水浴各时间组组间差异也均较显著(P<0.05)。对照组(37℃)细胞凋亡率为(5.92±0.87)%,55℃水浴3min组细胞凋亡率为(13.28±0.61)%,55℃水浴5min组细胞凋亡率为(50.82±3.72)%,55℃水浴8min组细胞凋亡率为(64.90±0.59)%。与对照组比较,55℃水浴各时间组细胞凋亡率均明显增高,且组间差异具有统计学意义(P<0.05),在温度恒定的前提下,表现为随着水浴时间的延长,兔RPE细胞凋亡率越来越高。透射电镜能明显看到细胞凋亡形态的改变。结论:55℃水浴加热3、5、8min均可造成兔RPE细胞凋亡,水浴时间越长,细胞凋亡率越高。

Objective To observe the effect of high temperature on cultured rabbit retinal pigment epithelial cells.Methods Rabbit retinal pigment epithelial cells were cultured in vitro by heating at 55 ℃ for 3 min, 5 min and 8 min.The cell viability was measured by CCK-8 method immediately after the treatment,cell apoptosis was determined by flow cytometry,morphological changes of cells were observed by transmission electron microscopy（TEM）.Results 55℃water bath OD value was significantly lower than the control group,the difference was statistically significant,and between 55℃ water bath each time group group differences was statistically significant difference （P〈0.05）.The control group （37℃） cell apoptosis rate was （5.92±0.87）%, 55℃ 3min group cell apoptosis rate was （13.28±0.61）%, 55℃ 5min group cell apoptosis rate was （50.82±3.72）%, 55 C 8min group cell apoptosis rate was （64.90±0.59）%.Compared with the control group, the apoptotic rate of the 55 ℃ water bath group was significantly higher than that of the control group （P〈0.05）.Under the constant temperature condition, with the prolongation of water bath time, Retinal pigment epithelial cell apoptosis rate is getting higher and higher.TEM can clearly see the changes apoptotic morphology.Conclusion The apoptosis of retinal pigment epithelial cells can be induced by heating at 55 ℃ for 3min, 5min and 8min, water bath time is longer, the higher the rate of cell apoptosis.