Extraction Of Active Components From Antler Velvet And Deer Fetus And A Study On Its Anti-senescence Effect

Antler velvet and deer fetus are rich in active ingredients,used as traditional anti-aging medicines with a wide range of pharmacological effects.There is a positive meaning for promoting the development of related functional products and deer industry,using of modern biology methods to research their anti-aging effects.In this study,we extracted the water and alcoholic active ingredients from the dry antler velvet,fresh antler velvet and deer fetus,respectively,and carried a comprehensive study of their anti-aging effects through such respects as measuring the toxicity and apoptosis of 3T3-L1 cells,the vitro antioxidant capacity,the life cycles of Caenorhabditis elegans and several related physiological and biochemical indicators of natural aging C57BL/6 mice.The results are as follows:1.Due to the fact that the vast majority of the active ingredients of antler velvet and deer fetus are water and alcoholic soluble,so we chose water and ethanol as extraction solvent.The extraction rate of freeze-dried water soluble substance of dry antler velvet were 3.944%,14.202%,45.652%,wherein the total protein contents were 28.87%,71.11%,58.21%,respectively.Detected by SDS-PAGE,its protein molecular weight distribution were in the range of 26-180 kDa.The alcoholic soluble extraction of three materials were purified by rotary evaporation under reduced pressure to obtain yield of 3.83%and 4.29%,11.44%,which the total phospholipid contents were 14.877 mg/ml,23.378 mg/ml,35.962 mg/ml.2.Considering that the toxicity of human plasma associated with vitro cytotoxicity,we detected the cytotoxicity of six groups of extracts to 3T3-L1 cells.The results showed that water soluble substance had no cytotoxicity when the concentration below 0.5 mg/ml,even the situation appears to promote cell proliferation,so did the alcoholic soluble extraction with less concentration of 0.1 mg/ml.Combined with flow cytometry to analyze cell apoptosis,we hypothesized that the extraction might have the potential to inhibit cell spontaneous apoptosis.3.To detect the vitro antioxidant capacity of antler velvet and deer fetus extracts,DPPH scavenging and CUPRAC law were applied.Antioxidant capacity of extracts exhibited a dose-dependent relationship in different concentrations.When chose 0.5 mg/ml of WFA and 0.1 mg/ml of EF as the positive groups,DPPH clearance rates were 42.17%,21.58%,and TEAC values were 325.56 ?M,113.36 ?M,respectively.4.Caenorhabditis elegans model was chose on researching the anti-aging effects of deer products extracts at the first time,and the study showed that 0.5 mg/ml of water soluble extract of fresh antler velvet and 0.1 mg/ml of ethanol soluble substance of deer fetus could prolong the life of the nematode significantly.The average and maximum lifespan of EF group were 25.16 days,34.36 days,while the WFA group were 26.64 days,36.44 days,respectively.In addition,the extracts two group could improve mobility of nematodes,also increased their capacity of resisting to environmental heat stress.Statistical results of nematode fecundity showed that the extracts might play a role in anti-aging through inhibiting reproductive capacity of nematode.We had the evidence to prove that the intervention of two groups extracts to life cycle of nematode sought to be independent of OD value of OP50.5.We detected changes in the serum or liver and brain related to aging indicators after natural aging C57BL/6 mice were fed the extracts of deer products for 21 days.Compared with the control group,the concentrations of T-CHO and TG in serum were reduced,so were the MDA content in serum,liver and brain tissue,while the vitality of SOD,GSH-Px and CAT had a different degree of improvement.The study found that anti-aging effects of water soluble extracts of fresh antler velvet and ethanol soluble extracts of deer fetus were most obvious.The mechanism might be related to these two extracts could improve the body's antioxidant capacity,promote cell proliferation,inhibit cell spontaneous apoptosis,regulate metabolism in vivo biochemical processes and so on.The study had laid a theoretical foundation for further researching on antiaging function of antler velvet and deer fetus,also provided a theoretical basis for the development of the relevant anti-aging products.