Peanut Skin Ameliorates Mice Thrombocytopenia Induced By Cyclophosphamide

Objective:Peanut skin is a common used Traditional Chinese Medicine in treating thrombocytopenia while actually the exact active component is still unknown. This study applied mice thrombocytopenia model induced by cyclophosphamide as a tool to screen the active fraction of peanut skin and trace its active component. In this way, we could understand what is in peanut skin really acts upon thrombocytopenia and primely study its rule in ameliorating thrombocytopenia.Method:Established a mice thrombocytopenia model induced by cyclophosphamide, and applied to found out an active fraction by analyzing the effects of peanut skin total extract, n-butanol fraction, ethyl acetate fraction and petroleum ether fraction on blood cells count, clotting time, bone marrow nucleated cell count, and bone marrow DNA content. The highly contained compound in this active fraction was then separated, and used mice thrombocytopenia model again to verify the pharmacological effect of this compound by analyzing blood cells count, bleeding time, and bone marrow DNA content. The mechanism of this compound acts on thrombocytopenia mice was studied by measuring blood cells count, bleeding time, thrombin-antithrombin complex content in sera, glutathione S-transferase, glutathione peroxidase, superoxide dismutase activity and malondialdehyde content in liver tissue and also by observing the histomorphology of liver.Result:Mice thrombocytopeina model was successfully established by continuous subcutaneous injection of 100 mg/kg cyclophosphamide for 3 days.This established mice thrombocytopeia model was used to screen the effect of peanut skin on platelet. The results demonstrated that peanut skin total extract medium dose (16g/kg, crude drug dose equivalent), high dose (32g/kg, crude drug dose equivalent), ethyl acetate fraction low dose (8g/kg, crude drug dose equivalent), medium dose (16g/kg, crude drug dose equivalent) could prevent thrombocytopenia and increase platelets count significantly. Peanut skin total extract low dose (8g/kg, crude drug dose equivalent) and high dose could decrease spleen index. Peanut skin total extract low, medium, high dose and ethyl acetate fraction high dose (32g/kg, crude drug dose equivalent), n-butanol fraction (32g/kg, crude drug dose equivalent) could decrease thymus index. Ethyl acetate medium dose could increase mice bone marrow DNA content.The pharmacodynamic test conducted to evaluate the effects of peanut skin ethyl acetate fraction and compound PNE-2 on thrombocytopenia mice showed that peanut skin ethyl acetate fraction low dose (8g/kg, crude drug dose equivalent), medium dose (16g/kg, crude drug dose equivalent), PNE-2 medium dose (120g/kg, crude drug dose equivalent) could raise white blood cells count; ethyl acetate fraction medium dose (16g/kg, crude drug dose equivalent), PNE-2 medium dose (120g/kg, crude drug dose equivalent), high dose (240g/kg, crude drug dose equivalent) could raise platelets count.But unfortunately, there was no platelet ameliorated effect of PNE-2 in the final experiment. Platelets, white blood cells count and thrombin-antithrombin complex content of model group mice dropped dramaticly, indicating a successful mice thrombocytopenia model. But PNE-2 showed no effect on those indexes. PNE-2 could prolong shortened bleeding time of thrombocytopenia model mice. Hepatic tissue glutathione S-transferase, glutathione peroxidase and superoxide dismutase activity in model group mice tended to be lower than those of control group, but with no statistically significance. There was no difference of malondialdehyde between groups.Pathological section of mice livers were stained with hematoxylin and eosin. Livers of model group mice damaged seriously, showing hepatocyte diffuse focal degeneration, cellular necrosis with karyorrhexis and inflammatory cells invasion. PNE-2 seemed to have no improvement effect on liver damage with cell degeneration and necrosis. But actually in PNE-2 treated groups, like A and C dose groups, Kupffer’s cells were seen all aroud while B dose group triggered inflammatory reaction vigorously.Conclusion:Peanut skin can remarkably ameliorate mice thrombocytopenia induced by cyclophosphamide. Peanut skin ethyl acetate fraction is the active fraction and PNE-2 separated from this fraction is the active compound. Its acting mechanisim might be related with its potentiation effect of immunological function, acceleration of hazard matter clearance, and promotion of platelet regeneration.