Studies On The Preparation,Identification Of CP Ⅰ And The Protective Effect Of CP Ⅰ To Cells Damaged By UVA

In this paper,how to prepare CP I and identify physico-chemical property had been studied, and UVA oxidative damage L929 was perliminary discussed. These datas provided theoretic foundation and scientific proof for further research and development of this product.Pigskin was used as material. According to the comparation among the three kinds of crude extraction methods including acid dissolve,enzymolysis and neutral salt solution. The method of enzymolysis combined with HPLC was chosen to prepare CP I .The physico-chemical property of ,CP I was identified. The results showed that: CP I showed character absorption peak at 223nm; CP I had 18 kinds of amino acids, total count of amino acid was more than 90%; the concentration of protein was 1. 03 mg/ml; CP I was composed of a chain, & chain and Y chain by SDS-PAGE; One pure peak with HPLC was founded and its purity was 100%. The security of CP I according to the experiments of mice accute toxicity was evaluated, Guinea pig sensitization and cytotoxicity experiment in integer level and cell level. It showed that : the mice had no toxic reaction; the Guinea pig's points of injection had no erythema and dropsy; the cytotoxicity was lower than I class. The results proved that CP I had no poison side effects.CP I prevented UVA from oxidative damaging L929 was studied. The activity ofcells became weaker and weaker with exposure time. It was conspicuous that UVA damaged the activity of cells. The contents of MDA upgraded and the contents of GSH-px degraded. When adding different concentration of CP I, the contents of MDA decreased and that of GSH-px increased. The effect of 50 % concentration group was extremely remarkable (P<0. 01). It showed that CP I can counteract oxidative damage from UVA.In conclusion, the method of enzymolysis combined with HPLC to extract CP I from pigskin was proposed. This method showed the adventages of high performance, high selectivity and easily amplification. Acoording to the experiments of identifying the physico-chemical property and the biosafety of CP I, it was concluded that CP I had high bioactivity and no toxicity effect, and that CP I had the ability to protect cells from oxidative damnification.