丹参素对LED光源诱导的人晶状体上皮细胞光损伤影响的研究

The effect of Salvianic acid A on the photic injury in hunman lens epithelial cells induced by LED light source

目的采用发光二极管（light emitting diode，LED）光源制作人晶状体上皮细胞（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 24h，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（MTY）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570nm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晶状体上皮细胞内的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。

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.