强脉冲光及5-氨基酮戊酸对小鼠B16黑素瘤细胞的作用

The Effects of Intense Pulsed Light and 5-aminolevulinic Acid on the Cultured B16 Murine Melanoma Cells

目的研究强脉冲光(IPL)及5-氨基酮戊酸(5-ALA)对小鼠B16黑素瘤细胞的影响。方法以小鼠B16黑素瘤细胞为实验对象,分为阴性对照组(不加5-ALA,未辐射)、UVA阳性对照组(3、4、5 J/cm2UVA辐射)、5-ALA组(含5 mmol/L 5-ALA的细胞培养液孵育)、IPL组(20、30、40 J/cm2IPL辐射)及5-ALA+IPL组(含5mmol/L 5-ALA的细胞培养液孵育细胞4 h后予20、30、40 J/cm2IPL辐射)。辐射后1 d、2 d、3 d、4 d、5 d以噻唑蓝比色法(MTT)测定细胞增殖情况、NaOH溶解法测定黑素含量、氧化多巴反应法测定酪氨酸酶活性的变化。结果5 mmol/L 5-ALA不影响B16黑素瘤细胞增殖、细胞黑素含量及酪氨酸酶活性。3～4 J/cm2UVA可促进B16黑素瘤细胞增殖,5 J/cm2时细胞增殖受到抑制。3～5 J/cm2UVA可提高酪氨酸酶活性、促进黑素合成,其作用呈剂量依赖性。IPL及5-ALA+IPL可降低黑素含量,5-ALA+IPL的作用较IPL更明显。IPL及5-ALA+IPL对细胞增殖及酪氨酸酶活性无明显影响。结论UVA可提高酪氨酸酶活性,促进B16黑素瘤细胞的黑素合成。IPL及5-ALA+IPL可使B16黑素瘤细胞黑素合成减少,对细胞增殖、酪氨酸酶活性无影响。

Objective To investigate the effects of IPL and IPL plus 5-ALA on the proliferation,melanogenesis and tyrosinase activity of cultured B16 murine melanoma cells.Methods B16 cells were pretreated with or without ALA and irradiated with IPL（20,30,40 J/cm^2）,also the negative control group and the positive control UVA irradiation group（3,4,5 J/cm^2） were designed.The proliferation of the treated B16 cells were evaluated with MTT method,the melanogenesis of the cells was tested with NaOH assay as well as tyrosinase activity was detected at 1 d,2 d,3 d,4 d,and 5 d after irradiation.Results 5 mmol/L 5-ALA did not affect the proliferation,melanogenesis and tyrosinase activity in B16 melanoma cell.The cell yielding in UVA group increased significantly（P0.05） compared with that of negative control group after UVA irradiation（3,4 J/cm^2）,but the proliferation of the treated B16 cells were suppressed after UVA irradiation（5 J/cm^2）.IPL（20,30,40 J/cm^2） or IPL plus ALA did not have any significant effects on cell proliferation.IPL or IPL plus ALA have inhibitory effects on melanogenesis without any influence on cell tyrosinase activity in B16 cells.ALA,as the photosensitizer,combined with IPL could have stronger effect than IPL alone.Conclusions We demonstrated that UVA could stimulate the proliferation of B16 cells,increase the melanogenesis and tyrosinase activity of the cells. IPL or IPL plus ALA have inhibitory effects on melanogenesis without any influence on cell proliferation and tyrosinase activity in B16 cells.IPL plus ALA had stronger effect than IPL alone.