摘要
目的 探讨组织工程心瓣膜常用人工基质材料--明胶发生降解后的产物对大鼠真皮成纤维细胞增殖及Ⅰ型胶原蛋白分泌的影响. 方法 利用酶组合技术制备明胶降解物（gelatin hydrolysate, GH）,采用超滤的方法分离出低分子量明胶降解物组分（gelatin hydrolysate fraction, GHF）,用于5只SD大鼠真皮成纤维细胞的体外培养.采用CCK-8活细胞计数试剂盒（Cell Counting Kit-8）检测细胞的增殖情况,ELISA试剂盒检测细胞I型胶原蛋白的分泌情况. 结果 ①GHF对细胞增殖作用的浓度效应：在0.125～1.0 mg/ml浓度范围内,GHF对细胞的促增殖作用呈现先增强后减弱的趋势,在0.25 mg/ml时,细胞增殖率最大,达到（47.54±16.35）%;②GH与GHF对细胞增殖影响比较：GH与GHF均具有促进细胞增殖的作用,且在浓度为0.25 mg/ml时,促细胞增殖作用无显著差异[（27.04±15.21）% vs （39.22±15.55）%, P=0.177];③GH与GHF对细胞分泌Ⅰ型胶原蛋白影响比较：GHF组第3天时Ⅰ型胶原分泌量为（28.06±5.60）pg/105,与空白对照的（18.24±4.52）pg/105相比,有显著差异（P=0.006）,而GH组直至第6天时Ⅰ型胶原蛋白分泌量,与空白对照相比,无统计学意义（P=0.103）. 结论 分子量大小不同的明胶降解物GH与GHF均具有促进大鼠真皮成纤维细胞增殖的作用,且分子量较小的GHF还具有促进细胞I型胶原分泌的功能.
Objective To investigate the influences of hydrolysate of gelatin, a commonly used artificial material for cardiac valve surgeries, on rat dermis fibroblasts proliferation and type Ⅰ collagen secretion. Methods Bio-enzymatic hydrolysis technique was used for obtaining gelatin hydrolysate （GH） , and ultra filtration for collecting the gelatin hydrolysate fractions （GHF） with lower molecular weight, which were applied in vitro culture of dermis fibroblasts from 5 SD rats. Cell proliferation and type Ⅰ collagen secretion were measured with Cell Counting Kit-8 and ELISA kit, respectively. Results （1）When the concentration fell in the range of 0. 125- 1.0 mg/ml, GHF could stimulate cell proliferation, especially at 0.25 mg/ml in which the proliferation rate reached the peak of （47.54 ± 16.35）%; （2）Compared with GHF, GH could also stimulate cell proliferation, and no significant difference was observed between them [（27.04±15.21）% vs （39.22 ±15.55）% , P=0.177] while the concentration was 0.25 mg/ml, even if GH had a higher molecular weight than GHF ; （3）The amount of type Ⅰ collagen secreted by rat dermis fibroblasts cultured in GHF group on the 3rd day was up to （28.06 ± 5.60）pg/105, significantly higher than the blank control （P = 0. 006）. However, no significant increase in type Ⅰ collagen secretion in fibroblasts cultured with GH was observed compared with the blank control group until the 6th day （P 〉0.05）. Conclusions Both GH and GHF, gelatin hydrolysates with various molecular weights, can stimulate rat dermis fibroblasts proliferation, and the latter can meanwhile increase type Ⅰcollagen secretion. Gelatin hydrolysate; Dermis fibroblast; Cell proliferation; Collagen secretion
出处
《中国微创外科杂志》
CSCD
2007年第8期815-817,共3页
Chinese Journal of Minimally Invasive Surgery
关键词
明胶降解物
真皮成纤维细胞
细胞增殖
胶原分泌
Gelatin hydrolysate
Dermis fibroblast
Cell proliferation
Collagen secretion
