近紫外线对兔晶状体上皮细胞Cx43蛋白影响的研究

Research of UVA in affecting Cx43 protein among rabbit lens epithelia cells

目的:探讨近紫外线抑制兔晶状体上皮细胞通讯功能的机制。方法:将兔晶状体前囊膜行组织块细胞培养,2～3代细胞分为正常对照组和近紫外线照射组(同一近紫外线光源下一固定位置50cm,测得该点的功率为0.2mW/cm2,给于同一强度,5、10、15min不同时间的近紫外线照射),应用Western-blot方法,从翻译水平检测近紫外线对兔晶状体上皮细胞(Rabbit lens epithelia cells,RLECs)的连接蛋白43(Connexin43,Cx43)表达变化的影响;采用间接免疫荧光染色法,对RLECs的Cx43蛋白进行定位分析。结果:Cx43蛋白在各组晶状体上皮细胞中均有表达,并且正常对照组为高表达,随着近紫外线照射时间的增加,其Cx43蛋白的表达逐渐降低;并经近紫外线照射后正常定位于晶状体上皮细胞膜上的Cx43发生了内移。结论:证实近紫外线抑制晶状体上皮细胞通讯功能的机制可能与细胞缝隙连接通讯(Gap junction-mediated intercellular communication,GJIC)的关键蛋白Cx43蛋白表达的下降和功能定位的改变有关。

Objective：To probe the mechanism of inhibition in gap junction-mediated intercellular communication （GJIC） of rabbit lens epithelia ceils （RLECs） irradiated by UVA. Methods：RLECs of 2 - 3 generation were divided into normal control group and ultraviolet irradiation group （The UV intensity was 0.2 mW/cm^2 under a UV lamp with fixed position 50 cm, RLECs were exposed to ultraviolet lamp at different times： 5,10,15 minutes）. Western blot techniques were employed to detect the protein levels of Cx43 irradiated by UVA. By indirect immunofluorescence histochemistry we analyzed the localization of Cx43 in the RLECs. Results：In UVA irradiation group and normal control group,there was expression of Cx43. With the increasing intensity,the expression of Cx43 was decreasing. The immunnostaining of Cx43 in RLECs revealed that the internalization of Cx43 from membrance to plasma and nuclear was found in UVA irradiated RLECs. Conclusion： Cell molecule level study proved UVA irradiation can reduce GJIC of RLECs, which wis associated with perturbations in localization and number of Cx43 among RLECs.