Department of Marine Bio-pharmacology, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China.
Department of Marine Bio-pharmacology, College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China.
Int J Biol Macromol. 2023 Aug 30;247:125772. doi: 10.1016/j.ijbiomac.2023.125772. Epub 2023 Jul 8.
Collagens from marine sources have been used widely in food, cosmetics and tissue engineering application due to their excellent functional and biological properties. In the present study, a novel protein, collagen from iris squid skin (SSC) was characterized, grafted with polyethylene-glycol (PEG) and Acid-Green 20 (AG) and was investigated the molecular signaling pathways in L-929 fibroblast cells along with their structural peptide analogs. SDS-PAGE and IR spectrum of SSC analysis showed the typical structure of type I collagen. The fibroblast proliferation was evaluated for SSC, SSC grafted PEG (SSC-PEG) and their structural analogs including Gly-Pro-Leu-Gly-Leu-Leu (PEP1), Gly-Pro-Leu-Gly-Leu-Leu-Gly-Phe-Leu (PEP2), Gly-Pro-Leu-Gly-Leu-Leu-Gly-Phe-Leu-Gly-Pro-Leu (PEP3) and Gly-Pro-Leu-Gly-Leu-Leu-Gly-Phe-Leu-Gly-Pro-Leu-Gly-Leu-Ser (PEP4). The optimal concentration of SSC and its derivative was 0.07 μ mol/L. The fibroblast growth-promoting factors were promoted by all the treatment groups by accelerating the PI3K/AKT and Ras/RAF/MAPK signaling pathways in L-929 cells, and inhibiting the secretion of apoptotic factors. Compared to the control group, mRNA and protein expression of AKT in the PI3K/AKT and Ras in Ras/RAF/MAPK signaling pathway were accelerated significantly by PEP4, respectively, while the Bax value was significantly lower (P < 0.01). The promoting effect of PEP1, PEP2, PEP3 and PEP4 on L-929 cells was closely related to the length of the peptides. Therefore, this study disclosed that PEP1, PEP2, PEP3 and PEP4 were novel analogs that greatly promote the proliferation of L-929 cells through PI3K/AKT and Ras/RAF/MAPK signaling pathways.
由于其出色的功能和生物学特性，来自海洋来源的胶原蛋白已广泛应用于食品、化妆品和组织工程领域。本研究中，对一种新型蛋白质——乌贼皮胶原蛋白（SSC）进行了表征，将其与聚乙二醇（PEG）和 Acid-Green 20（AG）接枝，并研究了其在 L-929 成纤维细胞中的分子信号通路及其结构肽类似物。SSC 的 SDS-PAGE 和 IR 图谱分析表明其具有典型的 I 型胶原蛋白结构。通过对 SSC、接枝 PEG 的 SSC（SSC-PEG）及其结构类似物（包括 Gly-Pro-Leu-Gly-Leu-Leu（PEP1）、Gly-Pro-Leu-Gly-Leu-Leu-Gly-Phe-Leu（PEP2）、Gly-Pro-Leu-Gly-Leu-Leu-Gly-Phe-Leu-Gly-Pro-Leu（PEP3）和 Gly-Pro-Leu-Gly-Leu-Leu-Gly-Phe-Leu-Gly-Pro-Leu-Gly-Leu-Ser（PEP4））进行了成纤维细胞增殖实验。SSC 及其衍生物的最佳浓度为 0.07μmol/L。所有处理组均通过加速 L-929 细胞中的 PI3K/AKT 和 Ras/RAF/MAPK 信号通路以及抑制凋亡因子的分泌来促进成纤维细胞生长促进因子的产生。与对照组相比，PI3K/AKT 信号通路中 AKT 的 mRNA 和蛋白表达以及 Ras/RAF/MAPK 信号通路中的 Ras 均显著加快（P<0.01），而 Bax 值显著降低（P<0.01）。PEP1、PEP2、PEP3 和 PEP4 对 L-929 细胞的促进作用与肽的长度密切相关。因此，本研究表明，PEP1、PEP2、PEP3 和 PEP4 是新型类似物，通过 PI3K/AKT 和 Ras/RAF/MAPK 信号通路极大地促进了 L-929 细胞的增殖。
