| It is widely recognized that Sophora subprostrate plays multiple roles in pharmacological and biological function as a traditional Chinese herbal medicine. The study was carried out to extract polysaccharides from Sophora subprostrate(SSP), work out the quality standard and investigate immuomodulatory function, free radical pharmacology of an effective constituent from the SSP. The results are as follows:Sophora subprostrate was treated in different periods, temperatures and solid-liquid ratio at microwave , the supernatants were treated with alcohol and freeze dehydration to obtain the polysaccharide, the extraction condition was 60℃, 90 min and 1:15 of solid-liquid ratio. the SSP yield was 5.54%. Drying shrinkage, monosaccharide, heavy metals, protein content all fit Pharmacopoeia of the People's Republic of China, The polysaccharides especial apices were seen by the infrared spectrum scan. The quality standard of SSP was also established.In order to investigate the immunomodulation and antioxidation functions of SSP, the mice were treated with dexamethasone (Dex,1.25 mg/kg) by subcutaneous injection On Day 1,3,5 alone or together with SSP by intraperitoneal injection at dose of 50, 100 or 200 mg/kg once daily for 7days .Chicken splenic lymphocytes were cultured in medium alone or with SSP at the final concentration of 50, 100, 200, 400μg/mL in vitro for 4, 8, 12, 24 h respectively to evaluate the effects of SSP on signal transduction, immune function and antioxidant ability on chicken. The supernatants at different culture periods were analyzed for levels of 6-keto-prostaglandin F1α(6-keto- PGF1α), thromboxane B2(TXB2) and nitric oxide. The cells were collected to determine contents of cAMP and cGMP. SSP at 50, 100 or 200 mg/kg were administrated by intramscular injection to chicken on Day 14. The spleen index, thymus index, bursa of fabricius index, SOD,MDA,T-AOC,GSH and NO in plasma were measured on Day 21, 28 ,35, 42 respectively. The results showed that SSP stimulated IFN–γsecretion of murine splenic lymphocytes at concentrations of 50, 100, 200 or 400 mg/L in vitro. SSP increased the levels of IL-6 and TNF-αin immunosuppressed mice induced by subcutaneous injection with dexamethasone at 1.25 mg/kg. SSP by intraperitoneal injection significantly raised spleen index, lysozyme activity in the immunosuppressed mice. SSP also decreased the level of IL-10 in mice serum. SSP significantly raised the activity of restrain hydroxy radical(·OH), content of hydrogen dioxide (H2O2) and level of reduced glutathione (GSH). The contents of nitric oxide (NO) was decreased. These findings suggested that SSP, a natural polysaccharide, may play an important role in regulating immunological functions in mice and possessing pronounced antioxidant activity. SSP at the final concentration of 100μg/mL increased the concentration of 6-keto-PGF1α, TXB2 and the production of NO in supernatants for 8 h culture. The concentration of intracellular cAMP was raised too. These results suggested that SSP was able to enhance immune function of the chicken splenic lymphocytes by regulating signal transduction mediated by arachidonic acid or cAMP/cGMP in chicken immunocytes. SSP could increase immune index and SOD, MDA, NO levels in plasma, T-AOC and GSH levels were raised then reduced. It suggested that SSP could regulate the immune function and antioxidant ability in chicken.ConclusionThis study developed a method to extract the polysaccharide from Sophora subprostrate, and this technique improved the yield of SSP. The quality standard of SSP was established. SSP may play an important role in regulating immunological functions and possess pronounced antioxidant activity in mice. SSP was able to enhance immune functions of the chicken splenic lymphocytes by regulating signal transduction mediated through arachidonic acid or cAMP/cGMP in chicken immunocytes and modulate the levels of reactive oxygen species in chicken.
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