Cao Yongqian, Wang Fagang, Jia Qingwei, Xu Rongjian, Dang Wei, Chen Qing, Lin Li, Wang Yibing
Department of Burns and Plastic Surgery, Provincial Hospital affiliated to Shandong University, Shandong, P R China.
Department of Orthopedics Affiliated Hospital of Taishan Medical Univisity, Shandong, P R China.
J Lasers Med Sci. 2014 Spring;5(2):75-81.
Pulsed dye laser (PDL) is an important treatment for superficial infantile hemangioma, but few studies report on its cellular mechanism. The aim of this study was to evaluate alterations of serum vascular endothelial growth factor (VEGF) level in infantile hemangioma (IH) patients after laser treatment and effects of PDL irradiation on human umbilical vein endothelial cells (HUVECs) in vitro, as well as to explore the biomolecular mechanisms and ultrastructure changes of the PDL effect.
74 children with infant hemangioma including 45 patients in proliferating phase, 18 patients in involuting phase, 11 patients in involuted phase and 10 healthy children were engaged in this study. The plasma VEGF levels of children were measured with the enzymelinked immunosorbent assay (ELISA). 24 hours after, HUVECs cultured in vitro were irradiated with PDL, cell apoptosis, mRNA levels of VEGF, and changes of ultrastructure were evaluated using flow cytometry, real-time reverse transcriptase polymerase chain reaction (RT-PCR), and transmission electron microscopy, respectively.
The serum VEGF concentrations in children with proliferating hemangiomas were significantly higher than in patients with involuting / involved hemangiomas and healthy patients. After receiving 3 laser treatments, the plasma VEGF levels of IH patients in proliferating hemangiomas decreased significantly. PDL irradiation could down-regulate VEGF mRNA expression of HUVECs, and increase cell apoptosis rate.
The present study demonstrates that PDL irradiation imparts apoptosis induction effects on HUVECs in vitro. Furthermore, our results suggest that vascular endothelial growth factor may be of particular importance in pathophysiology and PDL treatment of hemangiomas, also serum VEGF levels may be used as an aid in the follow up of IH. This provides valuable evidence of the PDL effect on infantile hemangioma.
脉冲染料激光(PDL)是治疗浅表性婴儿血管瘤的重要方法,但关于其细胞机制的研究报道较少。本研究旨在评估激光治疗后婴儿血管瘤(IH)患者血清血管内皮生长因子(VEGF)水平的变化,以及PDL照射对人脐静脉内皮细胞(HUVECs)的体外影响,同时探讨PDL作用的生物分子机制和超微结构变化。
本研究纳入74例婴儿血管瘤患儿,其中增殖期45例,消退期18例,消退完成期11例,另选取10例健康儿童作为对照。采用酶联免疫吸附测定(ELISA)法检测患儿血浆VEGF水平。体外培养的HUVECs在接受PDL照射24小时后,分别采用流式细胞术、实时逆转录聚合酶链反应(RT-PCR)及透射电子显微镜评估细胞凋亡、VEGF的mRNA水平及超微结构变化。
增殖期血管瘤患儿血清VEGF浓度显著高于消退期/消退完成期患儿及健康对照。接受3次激光治疗后,增殖期IH患儿血浆VEGF水平显著降低。PDL照射可下调HUVECs的VEGF mRNA表达,并增加细胞凋亡率。
本研究表明,PDL照射对体外培养的HUVECs具有诱导凋亡的作用。此外,我们的结果提示血管内皮生长因子在血管瘤的病理生理学及PDL治疗中可能具有重要意义,血清VEGF水平也可作为IH随访的辅助指标。这为PDL治疗婴儿血管瘤提供了有价值的证据。