摘要
目的 应用荧光淬灭后恢复 (fluorescenceredistributionafterphotobleaching,FRAP)技术研究兔晶状体上皮细胞胞间通讯 ,以及地塞米松和肝素对兔晶状体上皮细胞胞间通讯的影响。方法将体外培养兔晶状体上皮细胞接种于 96孔板 ,培养 2 4h后分别加入地塞米松 (浓度分别为 10、10 0、30 0mg/L)和肝素 (浓度分别为 1× 10 5、2× 10 5、4× 10 5U/L)。各组加入药物后 ,于 37℃继续培养 48h ,以羧基荧光素乙酰乙酸盐 ( 6 carboxyfluoresceindiacetate,CFDA)作荧光染色。以激光扫描仪 (ACASUltima)进行光淬灭并测定平均荧光恢复速率 (meanfluorescenceredistributionrate ,MFRR)。结果 地塞米松组 ,细胞内荧光淬灭后 ,平均荧光恢复速率 ( % /min)分别为 :对照组 0 375± 0 2 36 ,10mg/L组0 491± 0 2 39,10 0mg/L组 0 96 6± 0 44 7,30 0mg/L组 0 984± 0 379(F =2 6 0 7,P <0 0 5 ) ;肝素组 ,平均荧光恢复速率 ( % /min)分别为 :对照组 0 375± 0 2 36 ,1× 10 5U/L组 0 6 94± 0 491,2× 10 5U/L组1 0 97± 0 5 0 4,4× 10 5U/L组 1 0 82± 0 5 0 1(F =19 0 1,P <0 0 5 )。提示地塞米松和肝素可增强兔晶状体上皮细胞间的通讯 ,其作用与二者剂量有关。
Objective To investigate the intercellular communication in rabbit lens epithelial cells and effect of dexamethasone and heparin on it Methods Rabbit lens epithelial cells were cultured and exposed to different concentrations of dexamethasone (10, 100, 300 mg/L), heparin (1×105, 2×105, 4×105 U/L) for 48 hours at 37℃ The fluorescence redistribution after photobleaching (FRAP) was used for the analysis The mean fluorescence recovery rate (MFRR, %/min) of the cells labelled with 6carboxy fluorescein diacetate (CFDA) after photobleaching was measured with laser scanning cytometry ACAS Ultima Results MFRR (%/min) of the cells after exposure to dexamethasone was 0375±0236 in the control group, 0491±0239 in the group of dexamethasone 10 mg/L, 0996±0447 in the group of dexamethasone 100 mg/L, 0984±0379 in the group of dexamethasone 300 mg/L, respectively (F=2607, P<005) The MFRR after exposure of heparin was 0375± 0236 in the control group, 0694±0491 in the group of heparin 1×105 U/L, 1097±0504 in the group of heparin 2×105 U/L , 1082±0501 in the group of heparin 4×105 U/L, respectively (F=1901, P <005) Conclusion It is discovered that there is intercellular communication in rabbit lens epithelial cells with FRAP analysis Dexamethasone and heaprin can enhance the intercellular communication of rabbit lens epithelial cells in dose dependent manner
出处
《中华眼科杂志》
CAS
CSCD
北大核心
2000年第2期109-111,共3页
Chinese Journal of Ophthalmology
基金
中华眼科麦格科研基金资助项目!(97临01)
