关键词: 鹿茸多肽, 胶原蛋白, 细胞外调节蛋白激酶, 转化生长因子β1

Abstract: Objective: To investigate the effects of pilose antler polypeptide on the abilities of proliferation and collagen secretion of mouse embryonic fibroblasts NIH/3T3, and to clarify the relevant mechanisms.Methods: The NIH/3T3 cells were treated with different doses(1.56,3.13,6.25,12.50,25.00,50.00,100.00, and 200.00 mg·L^-1) of pilose antler polypeptide as experimental groups,the cells treated with 0 mg·L^-1 pilose antler polypeptide were used as blank control group, and the cells treated with 50.00 μg·L^-1 basic fibroblast growth factor(bFGF) were used as positive control group. MTT assay was used to detect the survival rates of NIH/3T3 cells in various groups. ELISA assay was used to detect the collagen secretion of NIH/3T3 cells in various groups.Wound healing assay was used to detect the migration abilities of NIH/3T3 cells.Western blotting method was performed to detect the expression levels of p-ERK 1/2 in the NIH/3T3 cells in various groups. Immunofluorescence method was used to detect the expression levels of transforming growth factor-β1(TGF-β1) in the NIH/3T3 cells in various groups.Results: Compared with blank control group,the survival rates of NIH/3T3 cells in positive control group and 6.25,12.50,25.00,50.00,100.00,200.00 mg·L^-1 pilose antler polypeptide groups were markedly increased (P<0.05 or P<0.01). Compared with blank control group, the levels of type Ⅰ collagen protein in the culture solution of the NIH/3T3 cells in positive control group and 6.25,12.50,25.00,and 50.00 mg·L^-1 pilose antler polypeptide groups were markedly increased (P<0.05 or P<0.01), and the levels of type Ⅲ collagen protein in the culture solution of the NIH/3T3 cells in positive control group and 12.50 and 25.00 mg·L^-1 pilose antler polypeptide groups were markedly increased (P<0.05). Compared with blank control group,the scratch healing rates of NIH/3T3 cells,and the expression levels of p-ERK 1/2 in the NIH/3T3 cells,and the expression levels of TGF-β1 in the NIH/3T3 cells in positive control group and 12.50 mg·L^-1 pilose antler polypeptide groups were markedly increased (P<0.05 or P<0.01).Conclusion: Pilose antler polypeptide can promote the proliferation, and collagen secretion of NIH/3T3 cells and increase the migration ability,which may be achieved by activating the phosphorylation of ERK 1/2 and increasing the expression of TGF-β1.

Key words: pilose antler polypeptide, collagen protein, extracellular signal-regulated kinase, transforming growth factor-β1