摘要
目的 探讨δ-氨基酮戊酸（ALA）光动力疗法（PDT）对瘢痕疙瘩成纤维细胞增殖、凋亡及胶原分泌的影响.方法 取体外培养的第3代瘢痕疙瘩成纤维细胞,给予0、1、3、6及9 mmol/L的ALA避光孵育3h,635 nm波长红光照射,能量密度为30 J/cm2,照射后继续孵育24 h,CCK-8法检测成纤维细胞增殖的抑制率;比色法检测羟脯氨酸含量;Western blot法检测丝氨酸/苏氨酸蛋白激酶Akt磷酸化激活形式p-Akt及程序性死亡因子4（PDCD4）的表达水平.结果 ALA在0、1、3、6及9 mmol/L的浓度下,瘢痕疙瘩成纤维细胞增殖的抑制率分别为0、（8.30±1.01）％、（29.48±3.27）％、（52.01±5.34）％、（79.99±5.85）％;羟脯氨酸含量分别是（9.540 0±0.352 42）、（6.242 5±0.224 85）、（5.107 5士0.534 88）、（3.4900±0.623 48）、（2.945 0±0.514 10） μg/mg;p-Akt的相对表达量分别为1、0.75±0.12、0.52±0.14、0.41±0.18、0.32±0.09;PDCD4的相对表达量分别为1、1.18±0.19、1.51±0.22、0.15±0.30、2.44±0.22.1、3、6及9 mmol/L组与0 mmol/L相比,差异均具有统计学意义（P＜0.05）.结论ALA在1～9 mmol/L范围内,均可可显著抑制成纤维细胞的增殖,减少羟脯氨酸的含量,表明δ-氨基酮戊酸光动力疗法可能是瘢痕疙瘩的一种潜在的治疗方法.
Objective To investigate the effect of δ-aminolevulinic acid （ALA） photodynamic therapy （PDT） on the cell proliferation,apoptosis and collagen secretion in keloid fibroblasts and to provide the theoretical base for ALA-PDT treatment of keloids.Methods Fibroblasts from keloid patients were cultured to the third generation in vitro and incubated in 0,1,3,6,9 mmol / L of δ-aminolevulinic acid for 3 h in the darkness.Then they were exposed to 635 nm wavelength red light （30 J/cm2） and continued incubation 24 h after irradiation.CCK-8 assay was used to detect proliferation inhibition rate of fibroblasts.The content of hydroxyproline was measured by colorimetric method.The expression of p-Akt and programmed cell death 4 （PDCD4） were detected by Western blot.Results The inhibition rate of keloid fibroblasts were respectively0,（8.30 ± 1.01） %,（29.48 ± 3.27） %,（52.01 ± 5.34） %,（79.99 ± 5.85） % with the presence of difference concentrations （0,1,3,6,9 mmol / L） of ALA.The content of hydroxyproline were respectively （9.540 0 ±0.352 42）,（6.242 5 ± 0.224 85）,（5.107 5 ± 0.534 88）,（3.490 0 ±0.623 48）,（2.945 0 ±0.514 10） μg/mg.The relative expression of p-Akt were respectively 1,0.75 ± 0.12,0.52 ± 0.14,0.41 ± 0.18,0.32 ± 0.09.The relative expression of PDCD4 were respectively 1,1.18 ± 0.19,1.51 ± 0.22,0.15 ±0.30,2.44 ± 0.22.The difference was statistically significant when compared the group of 1,3,6,9 mmol/L with 0 mmol/L（P 〈 0.05）.Conclusions In concentration within the range of 1-9 mmol/L,ALA could inhibit the proliferation of fibroblasts significantly,promote fibroblasts apoptosis and reduce the content of hydroxyproline in a dose-dependent manner,indicating that δ-aminolevulinic acid photodynamic therapy may be a potential treatment for keloid.
出处
《中华整形外科杂志》
CAS
CSCD
北大核心
2015年第1期49-53,共5页
Chinese Journal of Plastic Surgery
