[关键词]
[摘要]
目的:观察手机光照刺激体外培养人视网膜色素上皮细胞(retinal pigment epithelium; RPE)后形态及B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl2-Associated X(Bax)、半胱天冬酶-3(Caspase-3)的mRNA表达变化。
方法:将体外培养的人RPE细胞随机分成4组,根据RPE细胞光照时间长短分为照射3h光照组、6h光照组、12h光照组和0h光照组。在特制的培养箱内,将智能手机屏幕开到最大亮度(200±20Lx)做为光源,持续静音情况下循环播放彩色图片。然后应用苏木精-伊红(HE)染色法,末端脱氧核糖核酸转移酶介导的原位缺口末端标记(TUNEL)染色,四甲基偶氮唑盐微量酶反应比色(MTT)法观察各实验组RPE细胞形态学改变,并运用聚合酶链反应(PCR)技术检测各组细胞中Bcl-2、Bax、Caspase-3的mRNA表达变化。
结果:用HE染色、TUNEL染色、MTT法观察发现各光照组RPE细胞形态学无明显改变,差异均无统计学意义(P>0.05)。应用PCR技术检测发现RPE细胞在光照刺激3h光照组和6h光照组,细胞内的Bcl-2、Bax、Caspase-3的mRNA表达与0h光照组的差异均无统计学意义(P>0.05)。但随着光照时间持续延长(12h),细胞内的Bcl-2表达下调,Bax、Caspase-3表达水平均上升,差异均有统计学意义(P<0.05)。
结论:手机屏幕等作为人造光源在长期持续高亮度照射下会引起体外培养人视网膜色素上皮细胞的损伤。
[Key word]
[Abstract]
AIM: To research the morphological and fuctional change of human retinal pigment epithelium(RPE)induced by mobile phone screen light
in vitro and the mRNA express of apoptotic inhibitor B-cell lymphoma(Bcl-2), Bcl2-Associated X(Bax)and apoptotic actuators Caspase-3 in different groups.
METHODS: Cultured human RPE cells were divided into different group in random. According to the time of exposing, the four groups were 3h group, 6h group, 12h group and no light-exposed group. Cells of different groups were exposed to the opening mobile phone that was kept high-light brightness and playing on the colorful picture on silent mode. The morphological and functional change of RPE was quantified after exposed by HE staining, TUNEL staining, MTT, and use the technology of polymerase chain reaction(PCR)to detect the mRNA express of apoptotic inhibitor Bcl-2, Bax and apoptotic actuators Caspase-3 in different groups.
RESULTS: The different was not statistically significant(P>0.05)in the way of HE staining, TUNEL staining and MTT. The expression of Bcl-2, Bax and Capase-3 intracellular was no difference statistically significant in 3h, 6h group to no exposure group on the way of PCR(P>0.05). But as the time of exposed increased to 12h, the expression of apoptotic inhibitor Bcl-2 decreased, at the same time the expression of apoptotic actuators Bax and Caspase-3 increased, and the different was statistically significant(P<0.05).
CONCLUSION: The human RPE would be injured under the constantly high-light brightness of mobile phone screen as the man-made light source.
[中图分类号]
[基金项目]
武汉市卫生计生委科研基金资助项目(No.WX16E10)