Study On The Effects Of The MRNA Expression Of EPO, EPOR And GATA-1 Of The Marrow-suppressed Mice Treated By Three Kinds Of Therapies

OBJECTIVE:By detecting the effects to the level of peripheral blood RBC, HGB, PLT and hematopoietic cytokine EPO, EPOR and GATA-1 mRNA expression in the marrow gathered from Marrow suppression mice after chemotherapy in the ways of tonifying kidney, tonifying spleen, invigorate the circulation of blood with Liuweidihuang Tang,Buzhongyiqi Tang,Fufangdanshen Yin, probe into the therapeutical effect to Marrow suppression blood-insufficiency mice at molecular biology level, and then reveal the mechanism of promoting hematopoiesis of bone marrow suppression hemopenia mice in the ways of tonifying kidney,tonifying spleen,invigorate the circulation of blood to provide objective evidence and theoretical support for the Chinese medicine in prevention and treatment of making better choices of the ways of tonifying kidney,tonifying spleen,invigorate the circulation of blood to the patients with Marrow suppression Blood deficiency after chemotherapy.METHODS:100 mice were randomly divided into 5 groups, namely normal control group, model control group, Liu Wei Di Huang Tang group, Buzhong Yiqi Tang group and compound Danshen Decoction group, with 20 rats in each group. The mice were intraperitoneal injected after routine disinfection with 100mg/kg cyclophosphamide in the 9th,11th,13th days during the experiment.On the 14th and 28th, RBC,HGB and PLT in the conventional blood collected from bone marrow suppression mice model were tested by automated blood analyzer, while the mRNA expression level of EPO, EPOR and GATA-1 in the spleen were quantified by real-time fluorescence quantitative PCR.RESULTS:1. Compared with normal control group, the conventional blood RBC, HGB, PLT in the model control group were obvious decreased (P<0.05), showing that the model was successful. On the 14th day of the experiment, PLT was increased with statistically significant (P<0.05) in the Buzhongyiqi group, Liuweidihuang group, compound Danshen Decoction group compared with control group, while RBC and HGB were not. On the 28th day, RBC, HGB, PLT were significantly higher in the Buzhong Yiqi Tang group, Liuwei Dihuang Tang group and compound Danshen Decoction group with statistically significant (P<0.05) compared with the control group.2. On the 14th day of the experiment, compared with normal control group, the EPO and EPOR mRNA expression level in the model control group were little significant difference (P>0.05), while Liuwei Dihuang Tang group and Buzhong Yiqi Tang group significant difference and higher expression(p<0.01);Compared with model control group, the EPO and EPOR mRNA expression level of Liuwei Dihuang Tang group and Buzhong Yiqi Tang group were higher and the difference was significant(p<0.05). On the 28th day of the experiment, compared with normal control group, the model control group,Liuwei Dihuang Tang group and Buzhong Yiqi Tang group were significant difference with each other group(p<0.05); Compared with model control group, the EPO and EPOR mRNA expression level of Liuwei Dihuang Tang group and Buzhong Yiqi Tang group were higher expression and significant difference(p<0.05), while that has no statistical significance in the compound Danshen Decoction group and the level of EPO mRNA was influenced significantly by Liuwei Dihuang Tang.3. On the 14th day of the experiment, compared with normal control group, the GATA-1 mRNA expression level in the model control group was significant difference (P<0.05), while even higher in Liuwei Dihuang Tang group and Buzhong Yiqi Tang group and significant difference(p<0.01);Compared with model control group, the GATA-1 mRNA expression level in Liu Wei Di Huang Tang and Bu Zhong Yi Qi Tang were both increased and significant difference (P<0.05).On the 28th day of the experiment, compared with model control group, the GATA-1 mRNA expression level in the Liuwei Dihuang Tang group and Buzhong Yiqi Tang group were obvious higher and significant difference(P<0.05), while compound Danshen Decoction group with no statistical significance.CONCLUSION:1. Tonifying kidney,tonifying spleen,invigorate the circulation of blood can accelerate the recovery of the Peripheral Blood in the bone marrow suppression mice,and invigorate the circulation of blood Compound Danshen Yin can increase the RBC,HGB and PLT strongly.2. Liuwei Dihuang Tang and Buzhong Yiqi Tang both can accelerate the recovery of the Peripheral Blood in the bone marrow suppression mice through the promotion of EPO expression, and up-regulate the expression of GATA-1 and EPOR, and then GATA-1 up-regulate the expression of EPOR and GATA-1 proteins, thus the cells in the Peripheral Blood can get a better survival environment,proliferation and differentiation.3. Invigorate the circulation of blood Compound Danshen Yin can accelerate the recovery of the Peripheral Blood in the bone marrow suppression mice, not through improving EPO,EPOR and GATA-1 mRNA expression in bone marrow hematopoietic stem cells, but by improve the bone marrow micro-environment or other means to promote hematopoietic function to be restored, however the mechanism needs further study.