Effect Of Chinese Herbal Compound On Myelosuppressive Mice Hematopoiesis And Immune Function

Objective:To develop an efficient and low-cost Chinese herbal medicine immunopotentiator, the model of myelosuppression mice were established to confirm the efficiency of Chinese herbal compound(CHC) on hematopoietic and immune from biology, hematology, histology and immunology.Method:(1) Clean Kunming mice were selected in the trial by age of6-8weeks and quality range in18-22g.10mice (an equal split of♀and♂) in model group were taken intraperitoneal injection80mg/kg of cyclophosphamide at9:00a.m. one time per day for3consecutive days; an equal number of mice from control group were taken injections with an equal volume of physiological saline (0.9%wt/vol NaCl) by the same means. Blood samples were taken by removing the mouse eye and anticoagulated the bloodby EDTA-K2for the determination of blood physiological parameters at24hours post-injection. And the spleen and thymus were sampled for measuring the coefficient of organ.(2) There were130mice as described above were used to establish bone marrow suppression mouse model; and100of these mice were selected by spirit, body condition and divided into five groups (Ⅰ, Ⅱ， Ⅲ, Ⅳ, Ⅴ) by gender and quality,20mice (an equal split of♀and♂) in every group. Another20healthy mice (an equal split of♀and♂) were selected for the control group (Ⅵ). Mice in group Ⅰ, Ⅱ, Ⅲ were orally taken Chinese herbal compound respectively in doses (equivalent to crude drug content) of20g/kg,10g/kg,5g/kg; group Ⅳ were orally taken66mg/kg APS; Control group Ⅴ and Ⅵ were orally taken distilled water. The traditional Chinese medicine decoction was diluted with distilled water as dose of0.2ml (10g body quality) according to the corresponding dose of every group, and10mice(an equal split of♀and♂) in a group were given gavage at9:00a.m. one time per day for3consecutive days and another10mice for7consecutive days. Samples were taken at24h post-gavage the last time.(3) The blood physiological parameters and the mice bone marrow nucleated cells (BMC)were detected by Abacus Junior Vet Hematology Analyzer; and the reticulocytes (Ret) were counted with microscope by stained with1%brilliant cresyl blue salt.(4) The erythrocyte C3b receptors rosette and immune complex rosette was measured for evaluation the red cell immune function.(5) Serum immunoglobulins IgG, IgM, cytokines IL-2, IL-4, IFN-γ, IL-3, IL-6and EPO content were detected by ELISA method.(6) The pathology change of immune organ was observed by histological methods in spleen, thymus and bone marrow.Results:(1) The indexes of spleen and thymus and the content of BMC. WBC. GRA. LYM, PLT were decreased significantly in myelosuppressive mice; RBC, HCT and HGB were decreased slightly.(2) BMC in all mice were first increased and then decreased in the trial period, and BMC in the myelosuppressive mice changed significantly (P<0.01). BMC in group Ⅰ for orally taken CHC3consecutive days were increased significantly than that of the model group (P<0.01); and BMC in healthy control group for orally taken medication7consecutive days were increased significantly than that of the model group (P <0.01), and increased significantly than those of in group Ⅰ、Ⅱ and Ⅳ (P<0.05).(3) CHC groups (group Ⅰ, group Ⅱ, group Ⅲ) and astragalus polysaccharides could improve various types white blood cells, such as GRA, LYM and MID, and improve the GRA%and MID%, decrease LYM%; and increase Ret, RBC, PLT, increase the content of HGB in the bone marrow suppression mice. The dose of lOg/kg CHC for7days had best effectiveness on the bone marrow suppression mice.(4) The C3bRR of all mice were lower than those of before medication (P<0.01); and ICR of all mice changed slightly (P>0.05).(5) The content of IgG, IgM in all doses of CHC mice were increased after medication, and the effect of group II for7d was the best.(6) The content of IL-2in CHC group mice maintained at a high level, and the content of IFN-γ increased firstly and then decreased, and the reduction of IL-4and IL-4/IFN-γ were inhibited to a certain extent. The effect of group Ⅱ for7d was the best.(7) The contents of IL-3and EPO in CHC mice were increased, and the reduction of IL-6in the CHC mice were inhibited to some extent, the effect of middle dose CHC for7d was the best.(8) CHC can improve the clinical symptoms of the model mice and inhibit the decrease of the body weight. The quality and viscera index of model mouse spleen and thymus were increased in CHC group; and the pathological tissue morphology of the spleen, thymus and bone marrow were improved. The effect of middle dose medication for7d was the best.Conclusion:(1) The model of myelosuppression can be successfully established by daily intraperitoneal injection of80mg/kg cyclophosphamide for three days.(2) Chinese herbal compound can improve the hematopoietic function of myelosuppressive mice. The BMC, peripheral blood Ret, GRA, LYM, MID, RBC and PLT of the mice treated with Chinese herbal compound increased; hematopoietic cytokine IL-3and EPO and IL-6 concentration increased, the effect of10g/kg dose medication for7d was the best.(3) Chinese herbal compound can improve the immune function of myelosuppressive mice. It can resume C3bRR rate, make serum IgG, IgM, IL-2, IL-4content and IL-4/IFN-γ ratio increased, the spleen, thymus mass increased and the organ coefficient improved, promote the recovery of immune organ morphology.