Study On Structural Characterization Of Sika Deer Antler Polysaccharides And Their Effects On Liver Protection And Anti-aging In Vitro

The antler of sika deer is the young horn of sika deer（Cervus nipport Temminck）,which is rich in proteins,polysaccharides,and other substances.It has a wide range of physiological activities such as nourishing,antioxidant,and anti aging.The polysaccharide content of sika deer antler is rich,which is related to the efficacy of sika deer antler.Therefore,the purpose of this article is to analyze the structural characterization and anti aging activity of the extracted and purified sika deer antler polysaccharides,in order to clarify the pharmacological substances of the anti aging effect of sika deer antler,and also provide a reference for further research on sika deer antler polysaccharides.In this study,the total polysaccharide（SDAPO）of sika deer antler was obtained by hot water extraction and ethanol precipitation.Four components（SDAPO1,SDAPO2,SDAPO3 and SDAPO4）were separated by ion exchange chromatography（DEAE-52）and gel chromatography（Sephadex G-100）.The molecular weight,functional groups,monosaccharide composition,glycosidic bond configuration,and glycosidic bond bonding order of the four components were characterized and analyzed.SDAPO1,SDAPO2,SDAPO3,and SDAPO4 have typical polysaccharide structures with molecular weights of 1.57×10⁵,1.14×10⁵,9.85×10⁴and 7.92×10⁴Da,respectively.High performance liquid chromatography（HPLC）analysis showed that there were significant differences in the type and quantity of monosaccharides in SDAPO1,SDAPO2,SDAPO3,and SDAPO4.It is worth noting that compared to SDAPO2 and SDAPO3,SDAPO1 does not contain glucuronic acid and galacturonic acid,and SDAPO4 does not contain mannose,glucosamine hydrochloride,and galacturonic acid.Methylation analysis showed that the main polysaccharide fragments of SDAPO1,SDAPO2,SDAPO3,and SDAPO4 were→2,3)-Ribp（1→,→2,3）-Manp-（1→,→2,4）-Glc Ap-（1→,and→2,4）-Glc Ap-(1→.¹H NMR analysis showed that SDAPO1,SDAPO2,and SDAPO4 contained only one configurationβ-Pyranose,while SDAPO3 containsα-Pyranose andβ-pyranose.The results of this study described the structure-activity relationship between sika deer antler polysaccharides and their components,providing a reference for further research on sika deer antler polysaccharides.The scavenging activities of sika deer antler polysaccharide and its components against DPPH,ABTS,and hydroxyl radicals,as well as the total reduction ability against iron ions,were measured.The results showed that sika deer antler polysaccharide and its components had certain antioxidant activities in the concentration range of 1-10 mg/m L,among which SDAPO3 had higher scavenging activities against DPPH,ABTS,and hydroxyl radicals than other components of polysaccharides,with IC₅₀values of 4.60,2.10,and 3.20 mg/m L,respectively.The order of antioxidant capacity is SDAPO3>SDAPO2>SDAPO>SDAPO1>SDAPO4.The results of acute oral toxicity experiments in mice showed that there were no significant differences in body weight changes,organ index,and histopathological data analysis between the 8 g/kg dose of sika deer antler polysaccharide administered group and the blank control group（P>0.05）,indicating that sika deer antler polysaccharide is a safe and non-toxic natural antioxidant substance at the experimental dose.Screening alcoholic L02 hepatocyte injury models and exploring the effects of SDAPO,SDAPO1,SDAPO2,SDAPO3 and SDAPO4 on alcoholic hepatocyte injury.The results showed that the viability of L02 hepatocytes treated with 3%ethanol was55.87%,and this concentration of ethanol was selected as the alcoholic injury liquid for L02 hepatocytes.L02 hepatocytes were treated with sika deer antler polysaccharide and its components at 100-800μg/m L.The results showed that SDAPO1,SDAPO2 and SDAPO3 at 200μg/m L could promote the proliferation of L02 hepatocytes.The order of promoting proliferation was SDAPO3>SDAPO2>SDAPO1.Compared with alcohol injury model group,SDAPO2and SDAPO3 groups at 200μg/m L could significantly reduce alcohol-induced hepatocyte injury of L02（P<0.01）,the order of strength was SDAPO3>SDAPO2.In summary,the components of sika deer antler polysaccharide have an in vitro anti alcoholic liver injury effect,providing a theoretical basis for the study of sika deer antler polysaccharides’liver protective effect.In this study,we selected a model organism of Caenorhabditis elegans to study the anti aging ability of sika deer antler polysaccharides and their components.The results showed that compared with the blank control group,the treatment groups of sika deer antler polysaccharides（SDAPO,SDAPO1,SDAPO2,and SDAPO3）at three concentrations of 200,400,and 800μg/m L can extend the average lifespan of Caenorhabditis elegans,downregulate the ROS level in the nematode,and upregulate its swallowing and motor abilities,with the SDAPO3 administration group with a dose of 800μg/m L prolonged the average lifespan of Caenorhabditis elegans by27.01%,and had the best effect on prolonging the lifespan of Caenorhabditis elegans.To sum up,these research results show that the polysaccharide of sika deer antler has certain anti-aging activity,and also provides a reference for further research of polysaccharide of sika deer antler.The purpose of this study was to characterize the structure of Antinose Antinose polysaccharides and explore its antioxidant,liver protection and anti-aging effects in vitro.The results showed that each component contained mannose and uronic acid,which had free radical scavenging and reducing ability.SDAPO2 and SDAPO3 can reduce the liver cell injury of alcoholic L02,and have significant liver protection effect.SDAPO,SDAPO1,SDAPO2 and SDAPO3 can prolong the average life span of C.elegans,enhance its anti-stress ability,and have significant anti-aging activities.This study revealed the structural characteristics of sika deer antler polysaccharide,and proved its antioxidant,liver protection and anti-aging activities in vitro,which provided experimental basis for the study of physicochemical properties and pharmacological activities of sika deer antler polysaccharides.