| Potentilla anserine L. is a kind of plant which belongs to Rosacrae, Potentilla and is a typical stoloniferous clonal herbage in tibetan plateau, and potentilla anserina L. is the root tuber of Potentilla anserine L. used for Tibetan medicinal herb. Potentilla anserine L. polysaccharide (PAP) is one of its mainly active components from Potentilla anserine L. and have immunoregulation effects, so Potentilla anserine L. is popularly known in folk medicine for a variety of pharmacological effects, such as satisfy one's thirst, invigorating the spleen and benefiting the stomach, and tonifying Qi and nourishing blood. It was shown that crude extracts from Potentilla anserine L. had the effects on exhibiting immunomodulatory, anti-inflammatory and antibacterial activities, while it has been suggested that, among the active principles the water-soluble polysaccharides may greatly contribute to the biological activity of many Potentilla anserine L. extracts. The study was carried out to investigate the immunomodulatory effects of PAP. The mice peritoneal macrophages were collected, the phagocytotic function of mice peritoneal macrophages was measured with neutral red absorption test and the NO releasing by using Griess reagent; Neutrophilic leukocyte was collected, Reactive oxygen species (ROS) stimulated by opsonized zymosan (OZ), was detected by flow cytometry, and then the activity of myelopemxidase (MPO) measured; the immunosuppressive mice treated with Cy by intraperitoneal injection to investigate the immunomodulatory effects for PAP on the mice with PAP (50, 100, 200 and 400 mg/kg) or PAP (50, 100, 200 and 400 mg/kg) together with Cyclophosphamide (Cy, 100 mg/kg). The phagocytotic function of mononuclear phagocytic system was detected with carbon clear up methods, the levels of IL-6 and INF-γin serum were detected with ELISA,meanwhile, the activity of lysozyme in serum and the activity of the activity of SOD, GPx, CAT in spleen tissue were measured. The results were as follow:(1) PAP was able to potentialize phagocytotic function for peritoneal macrophage in a concentration-dependent manner, especially in the concentration of PAP 100μg/ml(P﹤0.01)in vitro.(2) PAP had promoted siginificantly the activities of MPO in a dose dependent manner, especially in the concentration of PAP 200, 400μg/ml(P﹤0.01)in vitro.(3) PAP could increasing the ROS production of neutrophil actived by Opsonized Zymosan (OZ) under Cy-induced immunosuppression by adjusting siginificantly the intensity of neutrophil respiratory burst.(4) PAP could siginificantly potentialize production of nitric oxide (NO) in vitro in a concentration dependent manner, especially in the concetration for PAP 200, 400μg/ml(P ﹤0.01)in vitro.(5) PAP could upgrate significantly the K value for clearance of carbon particles in mice treated with PAP (50,100,200 or 400mg/kg) alone in vivo, and increased significantly the K value for clearance of carbon particles in mice treated with Cy together with PAP (50, 100 or 200 mg/kg ).(6) PAP in the 4 concentration levels all of could siginificantly increased the activity of lysozyme in mice serum (P < 0.05).(7) PAP could promote the levels of IL-6 in serum, and PAP could overcome the immunological inhibition for cyclophosphamide so as to increase the level of IL-6 in the concetration of PAP 200, 400 mg/kg(P﹤0.01); PAP could promote the level of IFN-γin serum from mice treated with PAP (200 mg/kg) or PAP (200 mg/kg) together with Cy (100 mg/kg).(8) PAP had antioxidant effects in immune system by enhancing the antioxidant enzyme activity of SOD and CAT in vivo.In summary, the present reasearch demonstrated that PAP had the beneficial immunomodulatory activity, which could activate and/or prime macrophages and thus to enhance the modulation of phagocytosis activies, could also increase the production of NO from macrophage, enhance the activities of MPO and production of ROS from neutrophil. Moreover, PAP was able to raise the levels of IL-6 and IFN-γand lysozyme in serum, and had antioxidant effects in immune system by enhancing the antioxidant enzyme activity of SOD, CAT in vivo. It was suggested that PAP could enhance the immune defense function by modulating the immune function of immunocytes in vitro and Cyclophosphamide-induced immunosuppressed mice.
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