The Pharmacodynamics Evaluation Of Anoectochilus Roxburghii Active Ingredient

Objective:For many years, the tumor is one of insurmountable disease in the world.The traditional treatment is difficult to effectively prolong the life quality of patient. In recent years, traditional Chinese medicine(TCM) was proved to have good development prospect. Anoectochilus roxburghii is a representative. This experiment is intended to extract flavones, polysaccharide from Hedyotis diffusa, and verify the effects of the extract on tumor cells in vitro and the effects on immune function.Methods:Ethanol is used to extract flavonoids and obtain the powder by freeze-drying. For polysaccharide waterextraction and alcohol precipitation is the common method. The content is determined by UV. and cytotoxicity of HDW extract on A549 cells is measured by MTT assay.The phagocytic index and spleen coefficient on mice are study through lavage for 15 days. Get lymphocyte from mice spleen and treated with flavonoids and polysaccharide extract, Detecting the activity of cells by MTT assay then calculation proliferation rate of lymphocyte.Results:(1)1 g Anoectochilus roxburghii obtain 0.073 g crude flavonoids powder or0.031 g crude polysaccharide powder. Flavonoids content of flavonoids powder is 0.083mg·mg-1 and 6.082 mg·g-1 of the herb. Polysaccharide content of polysaccharide powder is0.117 mg·mg-1 and 3.667 mg·g-1 of the herb.(2)IC50 of Flavonoids extractive on A549 cells is189.204μg·mL-1, but polysaccharide don`t show up any inhibition.(3)Different doses(200, 400, 800 mg·kg-1·d-1) of flavonoids make different phagocytic indexs and spleen coefficiens. They are 0.034, 0.033, 0.030 and 6.468, 6.672, 6.836, respectively. The date of polysaccharide are 0.032, 0.040, 0.047 and 4.988, 4.664, 5.202.(4)Treated with different concentrations(40, 80, 160, 320 μg·mL-1) of flavonoids and polysaccharide on lymphocyte,proliferation rates of lymphocyte are 0.742, 0.770, 0.755, 0.186 and 0.252, 0.496, 0.534,0.347.Conclusions:(1)The contents of flavonoids and polysaccharide in Anoectochilus roxburghii are less than the datas in literature. Lacking extraction time, lossing material from transfer and reading error maybe cause that.(2)Flavonoids extractive can inhabit thegrowth of A549 cells and have a dose-dependent relationship. But polysaccharide extractive didn`t show any toxic, a specific amount of polysaccharide are helpful for tumo cells to grow.(3)Both flavonoids extractive and polysaccharide can enhance phagocytosis of macrophage, improve spleen coefficiens and promote growth of lymphocyte. The differents are that flavonoids extractive has stronger effect as a same concentration as polysaccharide extractive, but polysaccharide extractive has a better dose correlation.The reason is that high temperature make part of polysaccharide chain fracture when concentrate the water extracts.