本研究利用膠原酵素水解物(collagen enzyme-hydrolysed product complex, CEHPC)處理3T3老鼠纖維母細胞，分析細胞的生長及增生狀況、並偵測膠原蛋白第一型及第三型(collagen I 及collagen III) mRNA表現與細胞培養基內膠原蛋白的含量、同時進行膠原酵素水解物對基質金屬蛋白酶(MMP)-1、MMP-2及MMP-9的酵素活性抑制測定與其基因表現量的分析，在膠原酵素水解物的作用下，亦分析MMP抑制劑(TIMP-1、TIMP-2、TIMP-3及TIMP-4) 的基因表現量。本研究證明膠原酵素水解物在15%以內的濃度，處理72小時，對於3T3纖維母細胞均有80%以上的存活率；在10%的濃度下，於72小時內，與控制組相較可以促進膠原蛋白含量的增加，若處理纖維母細胞10~16小時，膠原蛋白的累積量可增加90%以上。因此，我們可以合理推測膠原酵素水解物(CEHPC)應具有促進真皮層纖維母細胞合成膠原蛋白的功效，可用於對抗細小皺紋，做為抗老化化粧保養品的有效成份。 This research employed collagen enzyme-hydrolysed product complex (CEHPC) to treat cultivated 3T3 mouse fibroblasts and analyzed cytotoxicity of CEHPC and the condition of cell proliferation. In this study, we detected mRNA level of collagen (collagen I and III), and determined the content of collagen present in medium. Furthermore, we investigated that the effects of CEHPC on activities and gene expression of MMPs (MMP-1, MMP-2, and MMP-9) and TIMPs (TIMP-1, TIMP-2, TIMP-3, and TIMP-4). The data presented here showed that 72 h treatment of fibroblasts in the culture medium with 15% of CEHPC could give rise to more than 80% survival rate of 3T3 fibroblasts, as well as 10% of CEHPC up-regulated the content of collagen in culture medium. By 10~16 h treatment with 10% of CEHPC, the accumulation of collagen increased more than 90% compared with the control. Therefore, we reasonably suggested that CEHPC has the capacity to promote fibroblasts to synthesize collagen. We thought that CEHPC can be utilized as an effective ingredient of anti-aging cosmetics for anti-wrinkling.
