Isolation and Purification of Sea Cucumber Peptide and Its Impact on the Secretion of Collagen by NIH/3T3 Cells
SONG Shu-liang
WANG Huan
LIANG Hao
JI Ai-guo
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.
Keywords：peptideproliferationcollagenNIH/3T3 cellsStichopus japonicas
Publication Date：2017-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：7( 22-28 )