CAJ | 학술논문

背景极低频电磁辐射产生的热效应与人类癌症的关系及其对眼表的影响已有较多研究.但眼球暴露于极低频电磁场(ELF-EMFs)下是否会导致巩膜的病理改变,从而影响近视的发生、发展目前鲜见报道. 目的研究ELF-EMFs作用下人胚胎眼巩膜成纤维细胞(HFSFs)的分子病理改变及其在近视发生发展中的可能机制.方法对HFSFs进行体外培养和传代,根据细胞是否暴露于50 Hz电磁场分为暴露组和对照组,应用实时定量聚合酶链反应( real-time PCR)法检测不同磁场强度(0、0.1、0.2、0.5、1.0 mT)、不同暴露时间(0、6、12、24、36、48 h)下HFSFs中Ⅰ型胶原蛋白(COL1A1)、基质金属蛋白酶2(MMP-2)的基因表达水平变化,同时应用CCK-8法检测两组HFSFs的细胞增生情况,并以细胞免疫荧光法对HFSFs中COL1A1和MMP-2蛋白的表达进行确认.结果与对照组相比,暴露组在磁场强度0.2 mT下作用6h可见HFSFs中COL1A1 mRNA的表达下调,差异有统计学意义(对照组:0.099±0.008,暴露组:0.050±0.004;P=0.009),且表达量随着磁场强度的增加而减少；在磁场强度0.1 mT下暴露24 h,HFSFs细胞中的MMP-2 mRNA表达上调,差异有统计学意义(对照组:0.009±0.001,暴露组:0.018±0.003:P=0.038),并随着暴露时间的延长表达增强.磁场强度0.2 mT下暴露24 h,HFSFs细胞增牛的吸光度(A450)值明显下降,差异有统计学意义(P=0.009);免疫荧光分析也显示暴露组HFSFs细胞中COL1A1表达下调,MMP-2表达上调.结论 ELF-EMFs暴露可在一定范围内影响体外培养的HFSFs的增生及COL1 A1的合成,可能是诱导巩膜发生病理性重塑进而导致近视发生、发展的危险因素之一.
배경 겁저빈전자복사산생적열효응여인유암증적관계급기대안표적영향이유교다연구.단안구폭로우겁저빈전자장(ELF-EMFs)하시부회도치공막적병리개변,종이영향근시적발생、발전목전선견보도. 목적 연구ELF-EMFs작용하인배태안공막성섬유세포(HFSFs)적분자병리개변급기재근시발생발전중적가능궤제.방법 대HFSFs진행체외배양화전대,근거세포시부폭로우50 Hz전자장분위폭로조화대조조,응용실시정량취합매련반응( real-time PCR)법검측불동자장강도(0、0.1、0.2、0.5、1.0 mT)、불동폭로시간(0、6、12、24、36、48 h)하HFSFs중Ⅰ형효원단백(COL1A1)、기질금속단백매2(MMP-2)적기인표체수평변화,동시응용CCK-8법검측량조HFSFs적세포증생정황,병이세포면역형광법대HFSFs중COL1A1화MMP-2단백적표체진행학인.결과 여대조조상비,폭로조재자장강도0.2 mT하작용6h가견HFSFs중COL1A1 mRNA적표체하조,차이유통계학의의(대조조:0.099±0.008,폭로조:0.050±0.004;P=0.009),차표체량수착자장강도적증가이감소；재자장강도0.1 mT하폭로24 h,HFSFs세포중적MMP-2 mRNA표체상조,차이유통계학의의(대조조:0.009±0.001,폭로조:0.018±0.003:P=0.038),병수착폭로시간적연장표체증강.자장강도0.2 mT하폭로24 h,HFSFs세포증우적흡광도(A450)치명현하강,차이유통계학의의(P=0.009);면역형광분석야현시폭로조HFSFs세포중COL1A1표체하조,MMP-2표체상조.결론 ELF-EMFs폭로가재일정범위내영향체외배양적HFSFs적증생급COL1 A1적합성,가능시유도공막발생병이성중소진이도치근시발생、발전적위험인소지일.
Background The effects of extremely low frequency electromagnetic fields (ELF-EMFs) on public health have attracted wide attentions.The association of the thermal effect of ELF-EMFs with cancer and ocular tissue damage has been of concern.However,the pathological changes of scleral tissue after exposure to ELF-EMFs as well as the relationship between these changes and myopia are still poorly understood. Objective The present study was to investigate the molecular pathological changes of human fetal scleral fibroblasts (HFSFs) after exposure to ELF-EMFs in vitro and to explore the possible mechanism in the occurrence and development of myopia.Methods HFSFs were cultured and passaged and then exposed to 50 Hz electromagnetic fields,and HFSFs that did not receive the irradiation of ELF-EMFs were used as the control group.The expression of collagen type Ⅰ (COL1A1 ) mRNA and matrix metalloproteinase-2 (MMP-2) mRNA in cultured HFSFs were detected by real-time qualitative polymerase chain reaction (real-time PCR) under different magnetic field intensites (0,0.1,0.2,0.5,1.0 mT) and different exposure time (0,6,12,24,36,48 hours).Cell proliferation assay of HFSFs was detected by the cell counting kit 8 ( CCK8 ) assay.The expression levels of COL1 A1 and MMP-2 proteins in HFSFs were further confirmed by immunofluorescence staining. Results The expression of COL1A1 mRNA was significantly down-regulated under the exposure of 0.2 mT ELF-EMFs for 6 hours,in comparison with the control group;moreover,it decreased in parallel with the increased of flux density (0.099±0.008 vs.0.050±0.004) (P =0.009 ).The expression of MMP-2mRNA was up-regulated conspicuously after exposure to 0.1 mT ELF-EMFs for 24 hours,and it increased with exposure time in comparison with the control group ( 0.009 ±0.001 vs.0.018±0.003 ) ( P =0.038 ).Proliferation of HFSFs (A450) was inhibited following the exposure to 0.2 mT ELF-EMFs for 24 hours in comparison with the control group (P =0.009 ).The expression of COL1 A1 in the experimental group was decreased,compared with the control group,but the expression of MMP-2 was increased. Conclusions ELF-EMFs inhibit the proliferation of HFSFs and expression of COL1 A1 in HFSFs,which might be one of the reasons for the development of myopia.