极低频电磁场对体外培养人视网膜色素上皮细胞的影响

Effects of extremely low frequency electromagnetic fields on human retinal pigment epithelium cells

目的 研究极低频电磁场（ELF-EMF）作用下体外培养的人视网膜色素上皮细胞（hRPE）的分子病理改变,探讨其在近视发生发展中的可能作用机制.方法 实验研究.根据细胞是否暴露于50 Hz电磁场分为暴露组和非暴露组,观察hRPE细胞形态的改变,CCK8检测hRPE细胞的增殖活性,应用实时定量聚合酶链反应（Real-time PCR）法检测hRPE细胞表达基质金属蛋白酶2（MMP-2）、基质金属蛋白酶组织抑制因子-2（TIMP-2）、转化生长因子-β2（TGF-β2）及成纤维细胞生长因子2（FGF-2）的情况.ELISA检测hRPE细胞培养上清中TGF-β2及FGF-2的表达情况,并用独立样本t检验进行分析.结果 与非暴露组相比,暴露组hRPE细胞形态发生改变;hRPE细胞的增殖活性降低,差异有统计学意义（6＆#215; 103/孔,t=-7.069,P＜0.01;8＆#215;103/孔,t=-3.652,P＜0.05;10＆#215;103/孔,t=-6.974,P＜0.05）;RT-PCR分析显示,暴露组hRPE细胞MMP-2、TIMP-2及TGF-β2 mRNA表达水平较非暴露组均增高,差异有统计学意义（t=10.562、6.277、4.940,P＜0.01）;而暴露组FGF-2 mRNA表达水平较非暴露组低,差异有统计学意义（t=-6.905,P＜0.01）.ELISA检测显示暴露组hRPE细胞上清中TGF-β2表达上调,FGF-2表达下降,与非暴露组相比差异有统计学意义（t=-4.079、4.441,P＜0.05）.结论 ELF-EMF在一定范围内影响体外培养的hRPE细胞的增生及相关细胞因子和基质金属蛋白酶的表达,ELF-EMF可能通过调控hRPE细胞病理改变进而诱导近视的发生发展.

Objective To investigate pathological changes in the molecules of human retinal pigment epithelium （hRPE） exposed to an extremely low frequency electromagnetic field （ELF-EMF）,and study its possible significance in the occurrence and development of myopia.Methods In this experimental study,the experimental group was composed of hRPE exposed to 50 Hz electromagnetic fields; the untreated group was composed of hRPE without exposure.The changes in hRPE cell morphology were observed and cell proliferation in hRPE was measured by a CCK8 assay.The expressions of matrix metallo-proteinase-2 （MMP-2）,tissue inhibitor of metalloproteinase 2 （TIMP-2）,transforming growth factor-β2 （TGF-β2） and fibroblast growth factor 2 （FGF-2） mRNA in hRPE were detected by reverse transcription-polymerase chain reaction （RT-PCR）.The concentrations of TGF-β2 and FGF-2 in the culture medium of hRPE were assayed by enzyme-linked immunosorbent assay （ELISA）.The difference between the two group using independent t test.Results hRPE cell morphology changed,and the proliferation of hRPE was inhibited （t=-7.069,-3.652,-6.974,P〈 0.05）.The mRNA expressions of MMP-2,TIMP-2 and TGF-β2 were up-regulated significantly （t=10.562,6.277,4.940,P〈0.01）.However,the mRNA expression of FGF-2 was down-regulated significantly （t=-6.905,P〈0.01）.The results of ELISA indicated that after exposure to ELF-EMF,the expression of TGF-β2 in hRPE supernatant medium was up-regulated significantly （t=-4.079,P〈 0.05）.However,the expression of FGF-2 in hRPE supernatant medium was down-regulated （t=4.441,P〈0.05）.Conclusion ELF-EMF changed the proliferation of hRPE,and also changed the expressions of MMP-2,TIMP-2,TGF-β2 and FGF-2 mRNA in hRPE,which might induce the occurrence of myopia through pathological changes in the molecules of hRPE.