CAJ | 학술논문

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.
목적 탐토유생소C화연선알대원대배양인표피세포적증식화분화적영향,위림상응용제공실험기출.방법 취정상인수술절제적포피,응용열용소분리표피여진피,이이단백매급EDTA분리표피세포위단개세포.이NIH-3T3작위사양세포,단개표피세포배양우표피완전배양기내.장배양적인표피세포등분위삼빈;일빈가유생소C,일빈가연선알,령일빈위완전배양액대조조.응용면역조화화단백질면역반점인적진렬기술검측C-myc、세포주기단백D1、사취합단백급내피단백적표체정황.결과 각조집락수불동,유생소C조＞대조조＞연선알조,유생소C조적집락형태여대조조근사,이여연선알조차이교대.여대조조상비,유생소C조C-myc、세포주기단백D1、사취합단백급내피단백적표체균상조(P치균＜0.05);연선알조사취합단백표체현저상조(P＜0.01),내피단백표체변동불명현(P＞0.05),이C-myc표체하조(P＜0.05).결론 유생색C대인각질형성세포적증식화분화균유촉진작용.연선알대인각질형성세포적분화유촉진작용,단대기증식각유억제작용.
Objective 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.