瘢痕组织中胶原蛋白提取及纯化研究

Extraction and Purification of Collagen from Scar Tissue

目的优化实验条件,建立提取瘢痕胶原蛋白的可靠方法。方法选择2015年1月~2016年12月本院20例手术整形切除增生性瘢痕的标本进行研究,用胃蛋白酶的消化方式、用盐酸溶解法提取瘢痕胶原,用氯化钠盐析法制成胶原蛋白膜,用吸收光谱分析、氨基酸成分分析对胶原蛋白进行鉴定。结果 20例胶原标本提取胶原蛋白,通过氨基酸成分分析甘氨酸占30%以上,吸收光谱分析在230 nm处有吸收波峰,证实提取物为来自皮肤瘢痕组织Ⅰ、Ⅲ型胶原蛋白。结论用酶消化、酸溶解、盐析法提取瘢痕胶原蛋白,方法可靠,操作简单,提取胶原蛋白纯度高,符合Ⅰ、Ⅲ型胶原蛋白特征,制成胶原蛋白膜为进一步研究瘢痕提供了生物学模型。

Objective To optimize experimental conditions and establish a reliable method for extracting scar collagen.Methods From January 2015 to December 2016,20 specimens of surgically resected hypertrophic scars were selected from the hospital.Pepsin digestion method was used to extract scar collagen by hydrochloric acid dissolution method,and collagen was prepared by sodium chloride salting out method.The protein film was identified by absorption spectroscopy and amino acid composition analysis.Results Collagen was extracted from 20 collagen samples, and glycine accounted for more than 30% by amino acid composition.Absorption peaks were observed at 230 nm by absorption spectrum analysis,and the extracts were confirmed to be collagen I and III from skin scar tissue.Conclusion The scar collagen was extracted by enzyme digestion,acid dissolution and salting out.The method was reliable,simple,and the purity of collagen extraction was high,which was consistent with the characteristics of type Ⅰ and Ⅲ collagen.The preparation of collagen membrane provides a biological model for the further study of scar.