蚕蛹活性肽调控LOVO细胞代谢的途径及构效关系研究

Silkworm pupa protein hydrolysates were proved with extensive activity on anti-tumor and, preliminary findings from our team indicated that the hydrolysates has a significant role to inhibit proliferation of LoVo cells, but its structure, anti-tumor molecular mechanisms and structure-activity relationship had been rarely reported. In the project, peptides of anti-tumor activity, which is from silkworm pupa protein, was prepared by controllable enzymolysis and sreened by activities on inhibit LoVo cells proliferation in vitro. Active peptide was isolated and purified by ultrafiltration and chromatography. The regulation on molecules expression levels of apoptosis family and related signal transduction pathway induced apoptosis in LoVo cells were studied. Moreover, the possible molecular mechanisms of antitumorigenic activities were elucidated. At the same time, Secondary structure, molecular weight, chain conformation and three-dimensional structure morphology structure characteristics in the solution were explored by using the multidimensional NMR, mass spectrometry, HPLC system and other advanced means. Quantitative structure-activity relationship (QSAR) theory was used to reveal structure-activity relationship on anti-tumor activity. This project provide theoretical and experimental basis for peptides of anti-tumor activity, which is from silkworm pupa, in the pharmaceutical and health care industry. It is important to utilize silkworm resources efficiently in China.

蚕蛹蛋白酶解产物具有广泛的抗肿瘤活性，项目组前期研究结果表明，蚕蛹酶解产物具有显著的抑制LoVo细胞增殖作用，但其具体的活性肽结构、作用机制及其构效关系仍需进一步研究，相关研究鲜有报道。本项目以蚕蛹蛋白为原料，拟可控酶解和体外LoVo细胞增殖抑制活性筛选制备目标活性肽，通过超滤、色谱层析等多种技术进一步分离、纯化蚕蛹抗肿瘤活性肽，从细胞凋亡途径切入，寻找影响LoVo细胞代谢的相关代谢途径，阐明可能的抗肿瘤分子机制。同时，采用多维核磁共振、高效液相系统、质谱等先进手段并结合定量构效关系理论，探讨抗肿瘤活性肽的结构、分子量、在溶液中的链构象以及立体三维结构形貌等结构特征，揭示其结构与抗肿瘤活性之间的构效关系。本项目不仅为蚕蛹蛋白制备抗肿瘤活性肽用于医药和保健行业提供理论和实验依据，也对高效利用我国具有独特优势的蚕蛹资源发展具有重要意义。

蚕蛹蛋白酶解产物具有广泛的抗肿瘤活性，本项目以蚕蛹蛋白为原料，获得抑制肿瘤细胞增殖的高活性肽段组分，并确定了活性肽段组分抑制肿瘤胞增殖活性与抗氧化指标的相关性，其中DPPH自由基清除能力与抑制肿瘤细胞增殖具有显著的相关性。在此基础上，明确了活性肽段组成与增殖抑制活性的构效关系，并通过研究活性肽段对MGC-803细胞的结构观察、细胞周期、凋亡途径相关因子的影响，基本阐明了SPPHs可能的抗肿瘤分子机制。本项目不仅为蚕蛹蛋白制备抗肿瘤活性肽用于医药和保健行业提供理论和实验依据，也对高效利用我国具有独特优势的蚕蛹资源发展具有重要意义。