猪Ⅱ型胶原及胶原肽抗关节炎的研究

Mechanism of Porcine Collagen Type Ⅱ and Collagen Peptide on Anti-arthritis

摘要:

本实验以猪鼻骨为原料，采用盐析法提取天然Ⅱ型胶原蛋白，采用碱性蛋白酶酶解制备胶原肽，并测定胶原蛋白的纯度，分析胶原蛋白及胶原肽的理化性质。通过构建小鼠CIA动物模型，研究Ⅱ型胶原蛋白及胶原肽对类风湿性关节炎的作用效果。结果表明，提取所得蛋白的胶原蛋白含量为82.86%，在紫外波长230 nm下有最大吸收峰，280 nm处几乎无吸收峰，分子量大于300KD，具备天然Ⅱ型胶原蛋白特征；酶解得胶原肽不含Ⅱ型胶原蛋白，分子量小于20 KD；Ⅱ型胶原蛋白能够降低小鼠关节指数、降低关节变形率，明显降低小鼠血清中IL-17、MMP-3和TNF-α水平（p<0.05），并能显著提高T细胞刺激指数（P<0.05），对B细胞刺激指数无明显效果（p>0.05）；组织病理学显示Ⅱ型胶原蛋白能够减轻关节破坏程度，抑制软骨退化；而胶原肽不具有以上作用。

Abstract:

Collagen was extracted from pig cartilage by salting out, and collagen peptide was hydrolyzed by alkaline protease. Collagen purity and the physicochemical properties of collagen and peptide were analyzed. The effect of anti-arthritic was studied by constructing CIA model. 82.86% of collagen was obtained, with molecular weight of more than 300 KD.The maximum absorption peak of UV was at 230 nm, and no obvious absorption peak was observed at 280 nm. Therefore the extracted collagen possessed the characteristics of natural collagen type Ⅱ, while the molecular weight of collagen peptide was less than 20 KD and no collagen type Ⅱ was observed. Collagen type Ⅱ could reduce index score and joint deformation rate, and significantly reduce IL-17, MMP-3 and TNF-α levels. In addition, Collagen type Ⅱ could significantly increase the T cell stimulation index (P＜0.05), but had no effect on B cell stimulation index (p＞0.05). Tissue sections showed that Collagen type Ⅱ could reduce destruction of joint and inhibit cartilage degradation. However, no such funcions were found with collagen peptide .