双波长脉冲染料激光抑制血管瘤内皮细胞增殖并促进其凋亡

Dual-wavelength pulsed dye laser inhibits proliferation and promotes apoptosis of hemangioma vascular endothelial cells

目的探讨双波长脉冲染料激光对血管瘤内皮细胞(HemECs)的增殖和凋亡的影响。方法体外分离培养HemECs并随机分为4组:脉冲染料激光组(PDL,595 nm波长)、Nd∶YAG激光(1064 nm波长)组、双波长组(595 nm PDL+1064 nm Nd∶YAG)、对照组(不照射激光)。于照射结束后第1、3、7天,采用CCK-8法检测各组HemECs的增殖能力,流式细胞术检测凋亡率,Western blot检测新生血管生成相关蛋白VEGF、bFGF的表达水平。结果激光照射后HemECs细胞较对照组细胞逐渐变长、变疏。照射后第1天,与对照组相比,双波长组HemECs的增殖抑制率显著升高(P=0.015),而PDL组、Nd∶YAG组的增殖抑制率均无明显改变(均P>0.05);照射后第3、7天,与对照组相比,PDL组、Nd∶YAG组及双波长组的HemECs的增殖活性、VEGF和bFGF蛋白表达水平均显著降低(均P<0.001),增殖抑制率与凋亡率均显著升高(均P<0.001),且双波长组较PDL组、Nd∶YAG组更明显(均P<0.05)。结论双波长脉冲染料激光可显著抑制HemECs增殖、促进其凋亡,降低血管生成因子水平,其作用较595 nm PDL或1064 nm Nd∶YAG单波长激光照射更明显。

Objective To investigate the effects of dual-wavelength pulsed dye laser on the proliferation and apoptosis of hemangioma vascular endothelial cells(HemECs).Methods The isolated HemECs were randomly divided into four groups:pulsed dye laser group(PDL,595 nm wavelength),Nd∶YAG laser(1064 nm wavelength)group,dual-wavelength group(595 nm PDL+1064 nm Nd∶YAG)and control group(no laser treatment).After 1,3,and 7 d of treatment,the proliferation and apoptosis rate of HemECs were detected using CCK-8 and flow cytometry,respectively.The expression levels of angiogenesis related proteins,VEGF and bFGF,were detected with Western blotting.Results Compared with the control group,laser treatment made HemECs longer and sparser.After one day of treatment,the dual-wavelength laser significantly inhibited HemEC proliferation(P=0.015).However,neither PDL nor Nd∶YAG significantly changed proliferation rate of HemECs(P>0.05 vs.controls).After 3 and 7 days of treatment,the proliferation activity,expression levels of VEGF and bFGF proteins of HemECs were markedly decreased in the all laser-treated groups(P<0.001 vs.controls),while the rates of proliferation inhibition and apoptosis were significantly increased(P<0.001 vs.controls),with a more prominent effect in the dual-wavelength laser-treated group(P<0.05 vs.PDL and Nd∶YAG groups).Conclusions Dual-wavelength pulsed dye laser can significantly inhibit the proliferation and promote apoptosis of HemECs,in addition to reduction in expression levels of angiogenesis factors.The effects of dual-wavelength pulsed dye laser are superior to that of either 595 nm PDL or 1064 nm Nd∶YAG single wavelength laser.