Study On Extraction Process Of Proanthocyanidins From Wild Pyracantha And The Anti-aging Effect Of Proanthocyanidins

Based on studies of proanthocyanidins , Spectrophotometry was used to optimizing the extraction process of proanthocyanidins from wild pyracantha fortuneana fruit ; Mouse skin fibroblasts was studied for its cell culture method.Then the skin fibroblasts was used as administration carrier to observe the effects of proanthocyanidins from wild pyracantha fortuneana fruit on cell culture.1.Response surface method was used to optimizing the extraction process of proanthocyanidins from wild pyracantha fortuneana fruit . On the basis of single-factor experiments and plackett-burman design successively.Box-Behnken center-united experimental design principles were used to devise the experiments and the response surfaces analysis of 3 factors,including water to material ration,ethanol concentration and pH of extracting solvent were adopted to further optimize extracting process.The results showed that the optimum parameters are:ratio of water to material is 12 to 1,ethanol concentration 73%, pH 3.6,extracting temperature and time,,80℃and 60min respectively,the yield of proanthocyanidin will arrive 93.446 mg/g under two times extraction.2.Mouse skin fibroblasts were cuitured by modified trypsin digestion method.Cell cultured needed 7days to get to more than 85% confluence.D-uring the subculturing.We can get a large of fibroblasts for the subsequent experiments.3.HE staining experiment was used to determine the cell morphology: cell which wo cultured were long spindle and polygonal. Cytoplasm was palered and nuclei blue.The results show that cultured cells were fibroblasts.4.Trypan blue staining was used to detect cell proliferation, cells began to grow from 2 days,and cells tend to balance after 5 days.It is 7.5×104个/mL.The results show that the growth of mouse skin fibroblasts complied with the general trend of cell growth. 5.The skin fibroblasts was used as administration carrier to observe theeffects of proanthocyanidins on cell proliferation ,activity of superoxide dis-mutase(SOD) and content of malondialdehyde(MDA) in cell.Treatment grou-p compared with the negative control group results showed that procyanidinscould promote mouse skin fibroblast cell survival and proliferation (P <0.001). and to promote the total intracellular SOD activity (P <0.001),r-eduction of MDA's Generation (P <0.01).In summary,Pyracantha can significantly improve fruit proanthocyanidins cell proliferation,can enhance the total intracellular SOD activity and impr-ove the ability of scavenging free radicals and reduce the MDA content of cells.So Pyracantha procyanidins on cell have some anti-aging effect.