CCK-8法检测蓝光和白光对ARPE-19细胞增殖的影响

目的：探讨蓝光和白光分别光照不同时间对人视网膜色素上皮(ARPE-19)细胞增殖的影响，为进一步检测光照损伤过程中相关因子的变化情况，研究光照损伤过程中的信号转导机制奠定基础。

方法：收集生长状态良好的ARPE-19 细胞进行实验，制作CCK-8标准曲线，确定实验合适的ARPE-19细胞密度及CCK-8试剂的反应时间。将细胞分为避光组、蓝光组、白光组，分别光照 3、6、9h，再避光培养12h后采用CCK-8 法检测不同光源光照不同时间对ARPE-19细胞增殖率的影响。

结果：通过CCK-8标准曲线选择每孔细胞数量为20 000个，CCK-8溶液作用4h后测量相应吸光度值。CCK-8检测结果示，避光组、蓝光组、白光组三组细胞光照不同时间细胞增殖率比较，差异有统计学意义(P<0.01)。蓝光组细胞光照3、6、9h，各时间点细胞增殖率比较，差异有统计学意义(P<0.001)，且随着光照时间的延长，细胞增殖率逐渐降低。白光组细胞光照3、6、9h，各时间点细胞增殖率比较，差异有统计学意义(P<0.05)，其中光照3h和6h细胞增殖率比较，差异有统计学意义(P<0.05)，而光照9h分别与光照3、6h细胞增殖率比较，差异均无统计学意义(P=0.253、0.120)。白光光照3～6h细胞增殖率呈下降趋势，而光照6～9h细胞增殖率呈现上升趋势。相同光照时间，蓝光组和白光组细胞增殖率均低于避光组，且蓝光组细胞增殖率均低于白光组，差异均有统计学意义(P<0.05)。

结论：光照能抑制ARPE-19细胞的增殖，其中蓝光光照细胞增殖率明显低于白光，且随光照时间的增长，细胞增殖率进一步降低。

Effect of blue light and white light on proliferation of ARPE-19 cells detected by CCK-8

AIM: To investigate the effects of blue light and white light on the proliferation of human retinal pigment epithelial cell line(ARPE-19)at different times, and lay a foundation for further detecting the changes of related factors during photodamage and further studying the signal transduction mechanism during light damage.

METHODS:Well-grown ARPE-19 cells were collected for experimentation. The standard curve of CCK-8 was made to determine the proper cell density of ARPE-19 cells and the reaction time of CCK-8 reagent. The cells were divided into dark group, blue light group and white light group, which were irradiated for 3, 6 and 9h respectively. After 12h of light-repellent treatment, CCK-8 method was used to examine the effects of different light sources on the proliferation rate of ARPE-19 cells at different times.

RESULTS: The number of cells per well was selected by CCK-8 standard curve to be 20 000, and the corresponding absorbance value was measured after 4h of the action of CCK-8 solution. The CCK-8 test results showed that the cell proliferation rates of the three groups were significantly different(P<0.01). The cell proliferation rate of the blue light group was significantly different(P<0.001)at 3, 6 and 9h, and the cell proliferation rate decreased gradually with the extension of the illumination time. The cell proliferation rate of the white light group was significantly different(P<0.05)at 3, 6 and 9h; there was a statistically significant difference in the rate of cell proliferation at 3h and 6h in white light(P<0.05), however, there was no significant difference in the rate of cell proliferation at 9h illumination compared with 3h and 6h illumination respectively(P=0.253, 0.120). The proliferation rate of cells under white light for 3-6h showed a downward trend, while that of cells under light for 6-9h showed an upward trend. At the same illumination time, the proliferation rate of the cells in the blue and white groups was lower than that in the dark group, and the cell proliferation rate in the blue group was lower than that in the white group. The difference was statistically significant(P<0.05).

CONCLUSION: The proliferation of ARPE-19 cells was inhibited by light irradiation. The proliferation rate of cells in blue light group was significantly lower than that of white light group. With the increase of light time, the cell proliferation rate decreased.