CAJ | 학술논문

目的采用发光二极管(light emitting diode,LED)光源制作人品状体上皮细胞(lens epithelial cell,LEC)光损伤模型,观察丹参素对LED光源诱导LECs光损伤细胞活性氧(reactive oxygen species,ROS)含量及凋亡的影响.方法实验研究.使用LED光源20000勒克斯(LUX)照射LECs及加入丹参素培养的LECs 24 h,使用流式细胞术检测LECs内ROS的表达及细胞凋亡情况,分别使用Real-time PCR和Western blotting检测凋亡相关基因BAX mRNA和蛋白表达,使用四甲基噻唑蓝(MTT)法检测细胞增殖.观察不同处理组间LECs在照射后细胞内以上各项指标的差异.结果光照组和丹参素光照组LECs细胞ROS平均荧光强度分别为15.83±5.81和7.67±0.63,细胞总凋亡率分别为(5.22±1.63)％和(1.57±1.21)％,光照组BAX基因的表达水平是丹参素光照组的(9.51±3.23)倍,以上检测指标两组间差异均有统计学意义(P＜0.05). 光照组、对照组和丹参素光照组BAX蛋白条带的校正后灰度值分别为0.297、0.065和0.100.MTT实验光照组与对照组和丹参素光照组,在570 nm处的吸收值A570分别为1.002±0.017、1.230±0.025、1.116±0.033,三组间比较差异有统计学意义(P＜0.01),各组之间比较差异均有统计学意义P ＜0.05).结论丹参素可以明显降低光损伤诱导的人品状体上皮细胞内的ROS水平,降低细胞凋亡的发生,丹参素对于光诱导的晶状体上皮细胞损伤的保护作用可能是通过清除细胞内的ROS来实现的.
목적 채용발광이겁관(light emitting diode,LED)광원제작인품상체상피세포(lens epithelial cell,LEC)광손상모형,관찰단삼소대LED광원유도LECs광손상세포활성양(reactive oxygen species,ROS)함량급조망적영향.방법 실험연구.사용LED광원20000륵극사(LUX)조사LECs급가입단삼소배양적LECs 24 h,사용류식세포술검측LECs내ROS적표체급세포조망정황,분별사용Real-time PCR화Western blotting검측조망상관기인BAX mRNA화단백표체,사용사갑기새서람(MTT)법검측세포증식.관찰불동처리조간LECs재조사후세포내이상각항지표적차이.결과 광조조화단삼소광조조LECs세포ROS평균형광강도분별위15.83±5.81화7.67±0.63,세포총조망솔분별위(5.22±1.63)％화(1.57±1.21)％,광조조BAX기인적표체수평시단삼소광조조적(9.51±3.23)배,이상검측지표량조간차이균유통계학의의(P＜0.05). 광조조、대조조화단삼소광조조BAX단백조대적교정후회도치분별위0.297、0.065화0.100.MTT실험광조조여대조조화단삼소광조조,재570 nm처적흡수치A570분별위1.002±0.017、1.230±0.025、1.116±0.033,삼조간비교차이유통계학의의(P＜0.01),각조지간비교차이균유통계학의의P ＜0.05).결론 단삼소가이명현강저광손상유도적인품상체상피세포내적ROS수평,강저세포조망적발생,단삼소대우광유도적정상체상피세포손상적보호작용가능시통과청제세포내적ROS래실현적.
Objective To investigate the Salvianic acid A on the effect on cleaning reactive oxygen species (ROS) and its anti-apoptosis effect of Light-emitting diode (LED) induced photic injury on lens epithelial cell (LEC) in vitro.Methods LED was used as the light source to induce the apoptosis of LEC in the study.The LECs with or without Salvianic acid A were exposed to LED light source during 24h with the illuminance of 20000 LUX.The expression of mean fluorescence intensity (MFI) of ROS and apoptosis in different groups were tested by flow cytometry (FCM).The expression of mRNA and protein of BAX in different groups were tested by Real-time PCR and Western blotting respectively.The proliferation of LEC of different groups was tested by methyl thiazolyl tetrazolium (MTT).Results The MFI of ROS in control group and Salvianic acid A group was 15.83±5.81 and 7.67±0.63,the total apoptosis rate of the two groups was (5.22±1.63)％ and (1.57±1.21)％,the expression of mRNA of BAX in the control group was 9.51 ±3.23 times higher than that in the Salvianic acid A group,the value of A570 of MTT was 1.002±0.017 and 1.116±0.033,the difference between the two groups was statistically significant (P ＜0.05).The expression of protein of BAX in the control group was 2.97 times higher than that in the Salvianic acid A group.Conclusions Salvianic acid A can decrease the cell apoptosis in the process of LED induced light damage of LEC which could realize by cleaning the of ROS expressed by the LEC in the process.