Studying Extracting Methods And Anti-aging Effect Of Hedysamn Polysaccharide On D-galactose Induced Aging Mouse Model

1. The Influence of Different Extracting Methods on Hedysamn Polysaccharide Antioxidation Activity in vitro.Objective:We screened for the extracting method from which Polysaccharide in high antioxidation activity in vitro obtained from the study of influence of different extracting methods on hedysamn polysaccharide antioxidation activity in vitro.Method:After preprocessing dry root of radix hedysari, the ultrasonic assisted, microwave-assisted and conventional hot water extraction methods were used to extract the polysaccharide, consequently, got HPS-C, HPS-M and HPS-H. Then, IR characterization scanning was used to determine the molecular weight of sample of HPS and the elimination ratio and reducing power of the removal of DPPH radical, hydroxyl radical and superoxide anion.Results:(1) Infrared spectrum analysis showed that the HPS-M, HPS-C and HPS-H extracted by the three different methods had absorption peak at the point of 3418cm-1,2935cm-1, 1618cm-1 and 1083cm-1, which were characteristic absorption peaks of polysaccharide, by which proving that the HPS-M, HPS-C and HPS-H are polysaccharide.(2) Determination of antioxidant activity in vitro experiments showed that the ability of HPS-M, HPS-C and HPS-H in removing hydroxyl radical, superoxide radicals and DPPH radical decreased in sequence, and antioxidant activity in vitro of the HPS-M is much higher than that of the HPS-C and HPS-H. At the same time, the HPS-M had the strongest reducing power (HPS-M>HPS-C＞HPS-H).Conclusions: Out of respect for hedysamn polysaccharides’bioactivity and extraction efficiency, microwave assisted as an effective method will be chosen for hedysamn polysaccharides extraction in the future.2. Anti-Immunosenescence effect of Hedysamn Polysaccharides on D-galactose Induced aging Mouse Model.Objective: D-galactose induced aging mouse model was establised to study the anti-immunosenescence effect of HPS.Method:Intraperitoneal injection of D - galactose was used to Kunming mice in clean grade to establish subacute senile mice model and were divided into 6 groups (n=10):Blank (normal saline group), model (D-galactose induced immunosenescence group), positive control (Vitamin E) and high, medium and low dose of hedysamn polysaccharides (HPS). Every group continuously accepted intraperitoneal injection of D-galactose (120 mg/kg) with exception of the blank group. The blank group was given saline, while the positive control group gave solution of vitamin E (50 mg/kg) and the three of high, medium and low dose of hedysamn polysaccharides group were given three different concentration solution (High dose group was given by 200mg/kg, medium dose group was given by 100mg/kg and low dose group was given by 50 mg/kg.), which lasted for 42 days continuously. Then the thymus index and spleen index were calculated. MTT was used to determine T-lymphocyte proliferation ability and T-lymphocyte subgroup level of CD3+T, CD3+CD4+T, CD3+CD8+T and CD3+CD28+T were assayed with flow cytometry. Determined the content of serum IL-2, IFN-γ, IL-4 and TNF-α in mice by ELISA.Results:(1) The model group obviously appeared aging signs. The various experiment data showed that the viscera index, T-lymphocyte proliferation and measurement of T cell subsets, levels of serum IL-2, IFN-γ, IL-4 and TNF-α of the model group were obviously different from that of the blank group.(2) High, medium and low dose of HPS group were all found the increasing thymus index and spleen index, which meant HPS had certain inhibition effect on immune organ atrophy of aging mice model and can restore T-lymphocyte proliferation ability, by raising the CD3+T cells, CD3+CD4+T cells and CD3+CD8+T cells in different levels in mice and increasing the number of the CD3+CD28+T cells obviously to regulate T lymphocytes.(3) Levels of serum cytokine IL-2, IFN-γand IL-4 in mice from the high, medium and low dose of HPS group have been increased significantly, but the expression of TNF-α was reduced in serum.Conclusions:High, medium and low dose of HPS group were all found the increasing thymus index and spleen index in subacute aging mice, which meant HPS had certain regulating effects on T-lymphocytes. HBS had certain anti-aging and immune regulation effects by effectively regulating T-lymphocytes to improve cellular immunity and humoral immunity function of sub-acute senile mice. Between the three doses of HPS groups, the anti-aging effect of the high and middle dose group were much obvious.