Study On Isolation And Anti-Aging Activity Of Antler Polysaccharide

Cornu Cervi Pantotrichum is the young horn of sika deer（Cervus nipport Temminck）or red deer（Cervus elaphus Linnaeus）,which is not ossified and densely hairy.The cornu Cervi Pantotrichum is sawed in summer and autumn,processed and dried in the shade or oven.Both male and female reindeer（Cervus rangifer Tarandus）have horns on their heads,which are mainly distributed in the Arctic region of the Northern Hemisphere and mostly grow in Daxing’anling region in China.Cornu Cervi Pantotrichum of reindeer antler,like that of sika deer and red deer,has high and similar medicinal value.Cornu Cervi Pantotrichum is a precious traditional Chinese medicine with a long history,which is mainly produced in Jilin and Liaoning.In recent years,because people’s research on cornu Cervi Pantotrichum mainly focuses on cornu Cervi Pantotrichum protein,the research on antler polysaccharide is not deep enough.Therefore,the purpose of this paper aims to extract and separate antler polysaccharide,and compare and analyze the differences of infrared spectra of the antler polysaccharide of sika deer（SDAPO）,reindeer（RDAPO）and red deer（RAPO）.What’s more,the structure of the antler polysaccharide components is characterized,and the antioxidant activity of the antler polysaccharide is studied.Caenorhabditis elegans is used as a model organism to study the anti-aging effect of the antler polysaccharide,which provides reference for finding new anti-aging drugs.In this paper,the hot water extraction method was used to extract antler polysaccharide.After degreasing,protein removal,pigment removal,dialysis and other operations,the SDAPO,RDAPO and RAPO were extracted.The extraction rates of SDAPO,RDAPO and RAPO measured by the phenol-concentrated sulfuric acid method were 3.38%,3.24%and 3.01%,respectively,of which the SDAPO content was the highest.Three components（SDAPO1,SDAPO2 and SDAPO3）were obtained by DEAE-52 ion exchange chromatography and Sephadex-G100 separation and purification.The purity of the components was acceptable by UV full wavelength scanning method,and they were collected for further analysis.Infrared spectroscopy combined with infrared second derivative method was used to analyze the functional group composition and differences of the three kinds of antler polysaccharide.The results showed that the functional group composition of SDAPO,RDAPO and RAPO was basically similar,but RDAPO and RAPO had a strong absorption peak at 1634 cm^-1,while SDAPO had a unique absorption peak at1618 cm^-1,and RDAPO had a broad absorption band at 1498 cm^-1.Therefore,it can be identified and provide reference for the identification and quality evaluation of antler polysaccharide from different deer.The structure of antler polysaccharide was characterized.The chemical analysis and structure characterization of SDAPO components were carried out by gel filtration method,Fourier transform infrared（FT-IR）spectroscopy,SEM and HPLC.The results showed that the three components had typical polysaccharide structure.Their molecular weights are 1.85×10⁵,1.54×10⁵ and 1.25×10⁵Da,respectively.Their surfaces are smooth and present irregular stereoscopic structures.SDAPO2 and SDAPO3 are composed of mannose,glucosamine hydrochloride,ribose,glucuronic acid,galacturonic acid,galactosamine hydrochloride,glucose and galactose.SDAPO1does not contain glucuronic acid and galacturonic acid.This study clarified the structure-activity relationship among the components,and provided reference for further study of antler polysaccharide.The antioxidant activities of SDAPO and total antler polysaccharide from different deer were determined in vitro,and DPPH free radical scavenging activities,ABTS free radical scavenging activities,hydroxyl free radical scavenging activities and total iron ion reducing power were determined respectively.The results showed that the SDAPO components and the total antler polysaccharide from different deer had antioxidant activity in vitro,and the antioxidant capacity was lower than that of Vc.Among SDAPO components,SDAPO3 has the lowest IC₅₀ value for scavenging free radicals and the strongest antioxidant activity,followed by SDAPO2,and SDAPO1 has relatively weak antioxidant capacity.The antioxidant activity of SDAPO is the strongest,followed by RAPO and RDAPO is the weakest among the total antler polysaccharide from different deer.This result provides a reference for the screening of natural active antioxidants.The anti-aging activities of SDAPO,RDAPO and RAPO were evaluated by using Caenorhabditis elegans as model organisms.The results showed that in the high dose group（800μg/m L）,SDAPO,RDAPO and RAPO groups prolonged the average life span of nematodes by 25.46%,12.70%and 15.69%,respectively.Among them,SDAPO has the best effect on prolonging the life span of Caenorhabditis elegans.Three kinds of antler polysaccharide all improved the swallowing and motor ability of Caenorhabditis elegans,but there was no significant change in oviposition ability.The SDAPO,RDAPO and RAPO groups significantly prolonged the life span of Caenorhabditis elegans under heat tolerance and oxidative stress（P<0.05）.Compared with the control group,the three antler polysaccharide groups had significant scavenging effect on ROS in Caenorhabditis elegans.These results indicate that the three kinds of antler polysaccharide have anti-aging effects,SDAPO has stronger anti-aging effect than RAPO,and RDAPO has weaker anti-aging effect.