MTT法和水溶性四氮唑(WST-1)法检测中波紫外线照射HaCaT细胞后活力的比较

目的:观察中波紫外线照射后HaCaT细胞活力的变化并探讨水溶性四氮唑(WST-1)法检测的适用性.方法:将HaCaT细胞分为对照组(假照射组)、300J/m2、600J/m2、900J/m2紫外线照射后24h组(每组6个样本).600J/m2紫外线照射后4h组、8h组、1 2h组、24h组、48h组、72h组(每组6个样本).血球计数板计算HaCaT细胞数量.四甲基偶氮唑盐(MTT)法和水溶性四氮唑(WST-1)法活性测定HaCaT细胞活性.结果:MTT法显示300J/m2、600J/m2、900J/m2紫外线照射HaCaT细胞24h后光密度(OD)值明显较0J/m2组低;WST-1法各组OD值明显比0J/m2组低.MTT法显示600J/m2紫外线照射HaCaT细胞8h、12h、24h、48h、72h后OD值明显比0J/m2组低.WST-1法显示600J/m2紫外线照射HaCaT细胞4h后OD值较0J/m2组低;600J/m2紫外线照射HaCaT细胞8h、12h、24h、48h、72h后OD值明显比0J/m2组低.WST-1法的OD值与细胞计数、MT法OD值呈正相关.结论:随着紫外线剂量的加大和照射后时间延长,HaCaT细胞数量减少,细胞的活力下降,WST-1代谢活性亦降低,呈时间和剂量依赖性.HaCaT细胞可对WST-1进行代谢,适用于该细胞增殖研究.

Comparison study on the viability of HaCaT cells after UVB irradiation by MTT and WST-1

Objective To observe the changes of viability of HaCaT cells after UVB irradiation and to investigate the applicability of WST-1.Methods HaCaT cells were divided into control group,300J/m2,600J/m2,and 900J/m2 UVB irradiation groups for 24 hours with 6 samples each group.600J/m2 UVB irradiation groups for 4,8,12,24,48,72 hours with 6 samples each group.HaCaT cell number was calculated with blood counting chamber and its activity was detected with MTT and WST-1 methods.Results OD values with MTT method with 300J/m2,600J/m2,and 900J/m2 UVB irradiation for 24 hours were significantly lower than that with 0J/m2 group.OD values with WST-1 method were significantly lower than 0J/m2 group.MTT methods show that:OD values with 600J/m2 UVB irradiation to HaCaT cells for 8,12,24,48,72 hours were significantly lower than 0J/m2 group.WST-1 methods showed that OD value with 600J/m2 UVB irradiation to HaCaT cells for 4 hours was lower than 0J/m2 group.OD values with 600J/m2 UVB irradiation to HaCaT cells for 8,12,24,48,72 hours were significantly lower than 0J/m2 group.The OD values of WST-1 methods positively correlated with cells counting and the OD values of MTT methods.Conclusion Number of HaCaT cells decreased and their activities became lower and metabolic activity of WST-1 became lower with the increase of UVB irradiation dosages and time,which shows time and dose-dependent.HaCaT cells may use WST-1 for metabolism and later may be suitable to study cell proliferation.