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  

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

作　　者：陈向齐[1] 刘向农[1] 曾抗[2] 兰海梅[2] 孙乐栋[2] 陈胜平[1] 牛高祥[1] 

机构地区：[1]福州总医院皮肤科,福建福州350025 [2]南方医科大学附属南方医院皮肤科,广东广州510515

出　　处：《中国美容医学》2007年第2期162-165,共4页Chinese Journal of Aesthetic Medicine

基　　金：南京军"区十五"科研基金(编号:02Z006)

摘　　要：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、12h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、MTT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。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.

关 键 词：MTT法 水溶性四氮唑(WST-1)法 中波紫外线 HACAT细胞 

分 类 号：Q813.1+1[生物学—生物工程]