摘要
背景:研究发现,珍珠液有促组织细胞再生、消除瘢痕等功效。目的:观察珍珠液对人皮肤瘢痕成纤维细胞增殖的影响。设计、时间及地点:分组对比观察实验,于2008-09/12在广西医科大学医学实验中心完成。材料:瘢痕组织组织标本来自广西医科大学整形外科患者。珍珠液是珍珠的水解液,含有多种氨基酸、多肽、维生素、矿物质和天然酶等,购自北海国发生物有限公司。方法:参考Veelken方法进行成纤维细胞原代及传代培养,反复传代去除非成纤维细胞成分后,建立人皮肤瘢痕细胞系。取对数生长期的人皮肤瘢痕成纤维细胞3～5代,用胰蛋白酶消化成单细胞悬液,接种于96孔塑料培养板,密度为0.5×104/孔,每孔加入100μL的细胞悬液和100μL的DMEM培养基,常规培养24h后吸弃原培养液,实验分组:珍珠液组:加入已经配置的含珍珠液质量浓度为125.00,62.50,31.30,15.60,7.80,3.90,1.95,0.98mg/L的培养基200μL;对照组:每孔只加200μL的培养基。主要观察指标:MTT法检测成纤维细胞增殖情况,TUNEL法检测成纤维细胞凋亡情况。结果:珍珠液对成纤维细胞的半效抑制浓度IC50为15mg/L,除0.98mg/L质量浓度外,其余质量浓度均能有效地抑制成纤维细胞的生长,且呈剂量依赖性抑制成纤维细胞的生长,珍珠液的浓度越高,对成纤维细胞的抑制率越大。剂量IC50=15mg/L培养的成纤维细胞体积缩小、胞质少、密度变小,凋亡细胞多、呈棕黄色。对照组成纤维细胞体积大、胞质丰富、密度大。珍珠液作用后的细胞凋亡指数高于空白对照组(P<0.01)。结论:珍珠液能抑制体外培养皮肤瘢痕成纤维细胞的生长,并诱导其凋亡。
BACKGROUND： Study shows that margarita liquid has effect on promoting histiocyte regeneration and removing scars. OBJECTIVE： To observe the effects of margarita liquid on the proliferation of fibroblasts in human skin scar tissues. DESIGN, TIME AND SETTING： A grouping contrast observational experiment was performed in the Experiment Centre of Guangxi Medical University from September to December in 2008. MATERIALS： Scar tissue samples were obtained from patients in the Department of Plastic Surgery, Guangxi Medical University. Margarita liquid was the digest of margarita pumhased from Beihai Gofar Marine Biological Industry Co., Ltd and rich in multi-amino acids, polypeptides, vitamin, mineral matters and natural enzymes. METHODS： Human skin scar cell line was established by removing non-flbroblasts through repeated primary culture and serial subcultivation of scar fibroblasts with reference to Veelken method. The 3-5 generation human skin scar fibroblasts on exponential phase of growth were made into single cell suspension by trypsin digestion which was then inoculated on plastic 96-well cell culture plate, with the density of 0.5×104/well as well as 100 pL cell suspension and 100 pL DMEM medium in each well. After culture for 24 hours, primary medium was discarded. The grouping of the experiment： 200 pL mediums with 125.00, 62.50, 31.30, 15.60, 7.80, 3.90, 1.95, 0.98 mg/L margarita liquid were used respectively in margarita liquid group; 200 pL pure medium was added into each well in control group. MAIN OUTCOME MEASURES： MI-F and TUNEL assay were used to examine the proliferation and the apoptosis of flbroblasts respectively. RESULTS： The 50% inhibiting concentration （IC50） of margarita liquid on fibroblasts was 15 mg/L. Margarita liquid at any other concentration but 0.98 mg/L was effective in inhibiting fibroblast growth in a dose-dependent way, Le. the higher margarita liquid concentration, the higher inhibition ratio on fibroblast growth. Fibroblasts cultured with 15 mg/L （IC50） margarita liquid had got reduced volume, lessened cytoplasm, decreased density, increased apoptosis rate and buffy colour. Fibroblasts in control group were large, rich in cytoplasm and compact. Apoptotic index was higher in the margarita liquid group than in the blank control group （P 〈 0.01）. CONCLUSION： Margarita liquid could inhibit the proliferation of skin scar fibroblasts cultured in vitro and induce the apoptosis of them.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2009年第46期9088-9091,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
广西自然科学基金资助项目(桂科青0447033)~~
