现代食品科技2017，Vol.33Issue(3)：22-28,7.DOI:10.13982/j.mfst.1673-9078.2017.3.004
海参肽的分离纯化及其对NIH/3T3细胞胶原蛋白分泌的影响
Isolation and Purification of Sea Cucumber Peptide and Its Impact on the Secretion of Collagen by NIH/3T3 Cells
摘要
Abstract
A sea cucumber peptide,SP12,was isolated and purified from Stichopusjaponicus,and its impact on NIH/3T3 cell proliferation and secretion of type Ⅰ collagen was studied.SP 12 was purified by ion exchange chromatography and gel filtration chromatography from fresh Stichopus japonicus using a cell activity tracking method.The effect of SP12 on NIH/3T3 cell proliferation was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay,and its impact on cell cycle progression was determined by flow cytometry.The effect of SP12 on collagen secretion by NIH/3T3 cells was measured by hydroxyproline kit,and its impact on the expression of type Ⅰ collagen,matrix metalloproteinase-1 (MMP-1),and tissue inhibitor of metalloproteinases-1 (TIMP-1) in NIH/3T3 cells was measured at the protein and gene levels by westem blot and reverse-transcription polymerase chain reaction (RT-PCR).The results clearly demonstrate that SP12 can considerably enhance the proliferation rate of NIH/3T3 cells and promote progression fiom G1 phase to S phase.In a range of 0～20 μg/mL,SP12 enhanced collagen secretion by NIH/3T3 cells in a dose-dependent manner.At the protein and genetic levels,SP12 considerably promoted the expression of type Ⅰ collagen and TIMP-1,and inhibited the expression of MMP-1;this might be the mechanism underlying SP12 promotion of collagen secretion by NIH/3T3 cells.关键词
多肽/增殖/胶原蛋白/NIH/3T3细胞/刺参Key words
peptide/proliferation/collagen/NIH/3T3 cells/Stichopus japonicas引用本文复制引用
宋淑亮,王欢,梁浩,吉爱国..海参肽的分离纯化及其对NIH/3T3细胞胶原蛋白分泌的影响[J].现代食品科技,2017,33(3):22-28,7.基金项目
国家自然科学基金资助项目(81371455) （81371455）
