RESUMO
BACKGROUND: Studies conducted on human cell culture models have demonstrated that collagen-derived peptides can exert a beneficial effect in medicine. However, all these studies were conducted using animal collagen samples, most often originating from bovine or porcine skin. Currently attempts are being made to replace animal collagen with fish collagen. OBJECTIVES: The aim of the study was to compare the effect of silver carp skin-derived peptide extract on the transcriptional activities of human VEGF and hsp70.1 gene promoters inserted into the plasmids with secreted alkaline phosphatase as a reporter gene. MATERIAL AND METHODS: Changes in the activity of the promoters were investigated using a HEK293FT cell line transfected with pVEGF-SEAP or pHsp70-SEAP. The cells were cultured in dishes containing peptides separated using reverse-phase high performance liquid chromatography. RESULTS: The study demonstrated that the silver carp skin-derived peptide extract exerts both an inhibitory effect on the VEGF gene promoter and activating effect on the hsp70.1 gene promoter. Higher biological activity was recorded in the case of a freshly prepared peptide extract compared to one stored at 4°C for three months. CONCLUSIONS: The silver carp skin-derived collagen peptides influence VEGF and hsp70.1 gene promoters' transcriptional activity.
Assuntos
Proteínas de Choque Térmico HSP70/genética , Peptídeos/farmacologia , Regiões Promotoras Genéticas/genética , Pele/metabolismo , Transcrição Gênica/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/genética , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Animais , Carpas , Colágeno/metabolismo , Relação Dose-Resposta a Droga , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Células HEK293 , Humanos , TransfecçãoRESUMO
BACKGROUND: Low-temperature techniques that prevent protein denaturation are being used to extract collagen from fish skin for cosmetic purposes. These extracts contain collagen with its triple helix structure preserved, as well as a number of other proteins. OBJECTIVES: The aim of the study was to investigate collagen extracts from the skin of silver carp for the presence of small-molecule peptides. MATERIAL AND METHODS: Liquid chromatography-mass spectrometry (HPLC-MS) was performed to analyze collagen extracts from silver carp skin for the presence of small-molecule peptides. RESULTS: A large number of different peptides were detected in the silver carp skin collagen extracts analyzed. Among the smaller peptides, the most abundant were those of 7-29 aminoacids originating from the following proteins: collagen Iα1, collagen Iα2, collagen Iα3, collagen VIα3, decorin, lumican, histone H2A, histone H2B and histone H4. CONCLUSIONS: The study demonstrated that, in addition to high-molecular-weight collagen proteins, acidic collagen extracts acquired from the skin of silver carp at temperatures up to 16°C also contain considerable amounts of small 7-29 amino-acid peptides. The application of these peptides could therefore be expected to result in beneficial clinical effects in patients in need of reconstructive treatment.