中华皮肤科杂志 ›› 2021, Vol. 54 ›› Issue (3): 232-235.doi: 10.35541/cjd.20190483

• 研究报道 • 上一篇    下一篇

激光照射对体外培养的婴儿血管瘤内皮细胞相关生长因子及细胞凋亡的影响

朱雅琳,万学峰,帕丽达．阿布利孜   

  1. 新疆医科大学第一附属医院皮肤科，乌鲁木齐  830054
  • 收稿日期:2019-04-10 修回日期:2020-02-24 发布日期:2021-03-02
  • 通讯作者: 帕丽达．阿布利孜 E-mail:Palidae@aliyun.com
  • 基金资助:
    新疆维吾尔自治区自然科学基金地区项目（2016D01C257）

Effect of laser irradiation on related growth factors in and apoptosis of in vitro cultured infantile hemangioma endothelial cells

Zhu Yalin, Wan Xuefeng, Palidae Abliz   

  1. Department of Dermatology, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, China
  • Received:2019-04-10 Revised:2020-02-24 Published:2021-03-02
  • Contact: Palidae Abliz E-mail:Palidae@aliyun.com
  • Supported by:
    Natural Science Foundation of Xinjiang Uygur Autonomous Region（2016D01C257）

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摘要： 【摘要】 目的 研究激光照射对体外培养的婴儿血管瘤内皮细胞相关生长因子及细胞凋亡的影响。方法 活化培养的婴儿血管瘤内皮细胞分为3组，强脉冲光（IPL）组（23 J/cm2，照射1次）、激光组（1 064 nm Nd：YAG激光，90 J/cm2，照射1次）和对照组（不照射激光）。照射后第1、3、7天，RT-PCR检测血管内皮生长因子（VEGF）、VEGF受体2（VEGFR-2）及碱性成纤维细胞生长因子（bFGF）mRNA的表达，Western印迹检测VEGFR-2蛋白表达，ELISA检测细胞上清液中VEGF和bFGF的含量，流式细胞仪检测细胞凋亡。结果 与对照组相比，1 064 nm Nd：YAG激光照射后第7天，VEGF mRNA（0.363 ± 0.021比1.000 ± 0.023）、VEGFR-2 mRNA（0.483 ± 0.017比1.001 ± 0.031）、bFGF mRNA（0.402 ± 0.040比1.000 ± 0.004）表达均下降，VEGFR-2蛋白表达下降（0.332 ± 0.055比0.768 ± 0.096），VEGF［（69.389 ± 24.179） ng/L比（334.506 ± 13.084） ng/L］和bFGF［（2.386 ± 0.151） ng/L比（9.165 ± 0.232） ng/L］分泌减少，细胞凋亡率（18.413% ± 2.654%比4.300% ± 0.036%）上升，差异均有统计学意义（P＜0.05）。与对照组相比，IPL组照射后第7天VEGF mRNA（0.436 ± 0.041比1.000 ± 0.023）、VEGFR-2 mRNA（0.493 ± 0.037比1.001 ± 0.031）、bFGF mRNA（0.490 ± 0.044比1.000 ± 0.004）表达下降，VEFGR-2蛋白表达下降（0.406 ± 0.037比0.768 ± 0.096），VEGF［（128.858 ± 6.063） ng/L比（334.506 ± 13.084） ng/L］和bFGF［（2.723 ± 0.471） ng/L比（9.165 ± 0.232） ng/L］分泌减少，细胞凋亡率上升（16.597% ± 1.877%比4.300% ± 0.036%），差异均有统计学意义（P＜0.05）。结论 1 064 nm Nd：YAG激光可能通过调节VEGF/VEGFR-2信号通路上的关键因子，以及细胞凋亡发挥对血管瘤内皮细胞的抑制作用，从而达到治疗血管瘤的作用。

关键词: 血管瘤, 激光疗法, 血管内皮生长因子类, 成纤维细胞生长因子, 细胞凋亡

Abstract: 【Abstract】 Objective To evaluate the effect of laser irradiation on related growth factors in and apoptosis of in vitro cultured infantile hemangioma endothelial cells. Methods Cultured infantile hemangioma endothelial cells were divided into 3 groups: intense pulsed light （IPL） group irradiated with IPL at a dose of 23 J/cm2 for 1 session, laser group irradiated with 1 064-nm Nd:YAG laser at a dose of 90 J/cm2 for 1 session, control group receiving no irradiation. On days 1, 3 and 7 after irradiation, RT-PCR was performed to measure the mRNA expression of vascular endothelial growth factor （VEGF）, VEGF receptor 2 （VEGFR-2） and basic fibroblast growth factor （bFGF）, and Western blot analysis to determine VEGFR-2 protein expression, enzyme-linked immunosorbent assay to detect the levels of VEGF and bFGF in the culture supernatant, and flow cytometry to detect cell apoptosis. Results Compared with the control group, the laser group showed significantly decreased mRNA expression of VEGF （0.363 ± 0.021 vs. 1.000 ± 0.023, P < 0.001）, VEGFR-2 （0.483 ± 0.017 vs. 1.001 ± 0.031, P = 0.001） and bFGF （0.402 ± 0.040 vs. 1.000 ± 0.004, P < 0.001）, decreased VEGFR-2 protein expression （0.332 ± 0.055 vs. 0.768 ± 0.096, P < 0.05）, decreased levels of VEGF （69.389 ± 24.179 ng/L vs. 334.506 ± 13.084 ng/L, P < 0.001） and bFGF （2.386 ± 0.151 ng/L vs. 9.165 ± 0.232 ng/L, P < 0.001） in the culture supernatant, but significantly increased apoptosis rate （18.413% ± 2.654% vs. 4.300% ± 0.036%, P < 0.01） on day 7 after irradiation. Compared with the control group, the IPL group also showed significantly decreased mRNA expression of VEGF （0.436 ± 0.041 vs. 1.000 ± 0.023, P < 0.05）, VEGFR-2 （0.493 ± 0.037 vs. 1.001 ± 0.031, P < 0.05） and bFGF （0.490 ± 0.044 vs. 1.000 ± 0.004, P < 0.05）, decreased VEGFR-2 protein expression （0.406 ± 0.037 vs. 0.768 ± 0.096, P < 0.05）, decreased levels of VEGF （128.858 ± 6.063 ng/L vs. 334.506 ± 13.084 ng/L, P < 0.001） and bFGF （2.723 ± 0.471 ng/L vs. 9.165 ± 0.232 ng/L, P < 0.001） in the culture supernatant, but significantly increased apoptosis rate （16.597% ± 1.877% vs. 4.300% ± 0.036%, P < 0.01） on day 7 after irradiation. Conclusion The 1 064-nm Nd:YAG laser may exert a therapeutic effect on hemangioma by inhibiting hemangioma endothelial cells via regulating key factors on the VEGF/VEGFR-2 signaling pathway and cell apoptosis.

Key words: Hemangioma, Laser therapy, Vascular endothelial growth factors, Fibroblast growth factors, Apoptosis