摘要目的 探讨维生素C和烟酰胺对原代培养人表皮细胞的增殖和分化的影响,为临床应用提供实验基础.方法 取正常人手术切除的包皮,应用热溶素分离表皮与真皮,以胰蛋白酶及EDTA分离表皮细胞为单个细胞.以NIH-3T3作为饲养细胞,单个表皮细胞培养于表皮完全培养基内.将培养的人表皮细胞等分为三份;一份加维生素C,一份加烟酰胺,另一份为完全培养液对照组.应用免疫组化和蛋白质免疫斑点印迹阵列技术检测C-myc、细胞周期蛋白D1、丝聚合蛋白及内披蛋白的表达情况.结果 各组集落数不同,维生素C组＞对照组＞烟酰胺组,维生素C组的集落形态与对照组近似,而与烟酰胺组差异较大.与对照组相比,维生素C组C-myc、细胞周期蛋白D1、丝聚合蛋白及内披蛋白的表达均上调(P值均＜0.05);烟酰胺组丝聚合蛋白表达显著上调(P＜0.01),内披蛋白表达变动不明显(P＞0.05),而C-myc表达下调(P＜0.05).结论 维生索C对人角质形成细胞的增殖和分化均有促进作用.烟酰胺对人角质形成细胞的分化有促进作用,但对其增殖却有抑制作用.

abstractsObjective To explore the effects of vitamin C and niacinamide on the growth and differentiation of human primary cultured keratinocytes.Methods Normal human foreskin was used in this study.The epidermis was separated enzymatically from the dermis by thermolysin,and keratinocytes were isolated from the epidermis by digestion with trypsin plus EDTA.The single keratinocytes were cultured with undedying NIH-3T3 cells as feeder cells in a complete medium supplied with 50 mg/L (vitamin C group),niacinamide of 400 μmol/L(niacinamide group)or vehicle(control group).Immunocytochemistry and immunodot blot were performed using monoclonal antibodies directed against C-myc,cyclin D1,filaggrin and involucrin.Results The colony number was highest in vitamin C group,followed by the control group and niacinamide group,and the colony morphology in vitamin C group was similar to that in the control group,but distinct from that in the niacinamide group.A significant increase was noticed in the expression of C-myc,cyclin D1,filaggrin and involucrin in vitamin C-treated keratinocytes compared with the control keratinocytes(all P＜0.05);however,in niacinamide-treated keratinocytes,the expression of filaggrin was significantly enhanced(P＜0.01),that of involucrin remained unchanged(P＞0.05),while that of C-myc was depressed(P＜0.05).Conclusions These results demonstrate that vitamin C has a favorable effect on both the growth and differentiation of human keratinocytes,while niacinamide seems to only promote the differentiation but attenuate the growth of human keratinocytes.