Effects Of Zuo Gui Bu Sui Sheng Xue Recipe On IFN-?/STAT1/T-bet Signaling Pathways In AA Mice Induced By Cyclophosphamide And Busulphan

Objective AA mice were induced by cyclophosphamide plus with busulfan,randomly divided into 5 groups: Control group,AA group,ZGBSSX recipe group(ZG group),Stanozolol group(ST group),ZGBSSX recipe combined with Stanozolol group(SG group).The following methods were used to observe the pharmacodynamic effects of the ZGBSSX and to explore its potential mechanism.Blood routine counts of white blood cell(WBC),red blood cell(RBC),blood platelet(PLT)and hemoglobin(Hb);Biopsy of bone marrow;Western blot was used to detect the expression of T-bet,STAT1,p-STAT1 and IFN-? protein in AA mice.Methods 100 specific pathogen-free ICR male mice,randomly divided into control group(n=20)and AA group(n=80).AA group: cyclophosphamide(40mg· kg-1· d-1),busulfan(20mg· kg-1· d-1),intraperitoneal injection in Day1,4,7,10 respectively.And then randomly divided the AA group into AA group(n=20),ZG group(n=20),ST group(n=20),SG group(n=20).The treatment groups continuous intragastric administration for 15 days;Control group and AA group uninterrupted intragastric administration of equal volume saline for 15 days.Extraction of blood from the medial canthus vein on day 15,20,25 and use the blood detect blood routine and ELISA.The general status and body weights of each group were recorded and observed during administration.At Day 25,all mice were killed.The right femur and spleen of each mouse were taken for pathological examination of bone marrow spleen tissue,to observe the pathological changes.The level of T-bet,STAT1,p-STAT1,IFN-? protein in the spleen tissues was detected in Western blot,to explore the mechanism of ZGBSSX recipe.The experimental data were statistically processed by SPSS19.0.Results1.General status: After the treatment of ZGBSSX,the mental state,appetite and fur glossiness of mice were improved.2.Body weights: Compared with the control group,the body weights of the AA group decreased significantly,with significant statistical difference(P < 0.05).The body mass of ZG group,ST group and SG group all increased in different degree(P < 0.05).3.Peripheral blood: Compared with the control group,the number of WBC,RBC,PLT,and Hb of AA group were all decreased significantly(P < 0.05).Compared with the AA group,the WBC,RBC,PLT and Hb of each treatment group increased to different degrees(P < 0.05)4.Biopsy of bone marrow Compared with the control group,AA group displayed reduced hematopoietic cells and abundant fatty cells,the proliferation of the bone marrow mononuclear cells(BMNC)was extremely decreased,the RBC,WBC and the megakaryocyte decreased to a different extent.Compared with AA group,all treatment groups had different degrees of recovery,fatty cells decreased obviously,RBC,WBC,megakaryocyte raised in varying degrees.5.Western blot The expression of T-bet,IFN-?,p-STAT1 protein in the AA group was significantly higher than that of the control group,and the expression of STAT1 protein was lower(P<0.01).Compared with the AA group,the expression of T-bet,IFN-?,p-STAT1 protein decreased,and the expression of STAT1 protein increased(P<0.01).Conclusion 1.As the results showed in this study we can conclude that the ZGBSSX can improve the general status of AA mice model,increase the body weight,reduce the pathological damage to bone marrow and spleen tissue,increase the number of RBC,WBC,PLT and Hb in peripheral blood,increase the number of BMNC.SG group is better than ZG group and ST group.2.It is possible that ZGBSSX improve the hematopoietic function of the bone marrow of AA mice by regulating IFN-?/STAT1/T-bet related signaling pathways.This study provides an experimental basis for the treatment of AA by TCM.