5-氨基酮戊酸联合光动力疗法可显著抑制内皮细胞生长并诱导细胞凋亡

Aminolevulinic acid photodynamic therapy can significantly inhibit the growth of endothelial cells and induce cell apoptosis

背景:增生性瘢痕目前缺乏安全有效的治疗方法。增生性瘢痕形成的关键点在于成纤维细胞过度增殖、血管内皮细胞增生及新生血管形成。有研究表明,激光连续照射可会诱导成纤维细胞凋亡。光敏剂5-氨基酮戊酸联合光动力疗法已广泛运用于许多临床肿瘤及皮肤疾病治疗,为治疗增生性瘢痕提供新方法。目的:观察5-氨基酮戊酸联合光动力疗法对于诱导内皮细胞凋亡的影响。方法:体外培养人脐静脉内皮细胞,实验分为4组,对照组无处理,单加药组加入5-氨基酮戊酸2 mmol/L,单照光组采用光动力3 J/cm^2照射8 min,加药照光组加入5-氨基酮戊酸2 mmol/L孵育4 h后再用光动力3 J/cm^2照射8 min。避光培养24 h。结果与结论:(1)荧光显微镜下观察:发现人脐静脉内皮细胞内荧光累积量呈5-氨基酮戊酸浓度依赖性,并在浓度为2 mmol/L时达到峰值;(2)CCK-8细胞毒性实验显示:单加药组细胞活性无显著差异。加药照光组细胞活性在药物浓度为2,4 mmol/L时受到显著抑制(P <0.01);(3)Hoechst 33258染色和流式细胞分析显示:5-氨基酮戊酸浓度为2 mmol/L时,5-氨基酮戊酸联合光动力疗法可以诱导内皮细胞出现凋亡(P <0.05);(4)结果证实,5-氨基酮戊酸联合光动力疗法可显著抑制内皮细胞生长,诱导内皮细胞发生凋亡。

BACKGROUND:There is a lack of safe and effective method for hypertrophic scar.Fibroblast hypertrophy,vascular endothelial cell hyperplasia and neovascularization are key points for hypertrophic scar formation.Continuous laser radiation has been shown to induce fibroblast apoptosis.Aminolevulinic acid photodynamic therapy(ALA-PDT)has been extensively applied in the treatment of tumor and skin diseases,which provide novel therapy for hypertrophic scar.OBJECTIVE:To explore the effect of ALA-PDT on endothelial cell apoptosis.METHODS:Human umbilical vein endothelial cells were cultured in vitro,and divided into four groups:control group(no treatment),single-drug group(incubation with 2 mmol/L 5-aminolevulinic acid),single-light group(3 J/cm2 photodynamic irradiation for 8 minutes)and ALA-PDT group(after incubation with 2 mmol/L 5-aminolevulinic acid for 4 hours,irradiated by 3 J/cm2 photodynamic for 8 minutes,and cultured for 24 hours in dark).RESULTS AND CONCLUSION:Fluorescence microscopy showed that the accumulation of fluorescence in umbilical vein endothelial cells was ALA-dependent and peaked at a concentration of 2 mmol/L.Cytotoxicity test showed no significant difference in cell viability in the simple-drug group.Cell viability in the ALA-PDT group was significantly inhibited at a drug concentration of 2 and 4 mmol/L(P<0.01).Hoechst 33258 staining and flow cytometry analysis showed that ALA-PDT could induce cell apoptosis when ALA concentration was 2 mmol/L(P<0.01).These results indicate that ALA-PDT can significantly inhibit the growth of endothelial cells and induce apoptosis of endothelial cells.