| Objective:The aim of this study was to explore the effects of Bushen Jianpi Quzhang Granules on Relationship between the ratio of Th1/Th2 and IFN-γ/T-bet and IL-4/STAT6 signalling pathways,T-bet,GATA-3,and the cytokines IFN-γ,TNF-α,IL-2,IL-4 and IL-10 expression in aplastic anemia model mice as well as investigating the possible underlying mechanisms of aplastic anemia of Thl/Th2 and the treatment of Bushen Jianpi Quzhang Granules,to provide theoretical evidence and support of Clinical treatment of aplastic anemia.We observe the clinical effect of "tonifying kidney invigorating spleen and removing barrier granules" on the treatment of aplastic anemia on the basis of previous research.Then observed the changes of baseline data,bone marrow hyperplasia,WBC,HGB,PLT,treatment efficiency,blood routine and adverse reactions in three groups of subjects,in order to evaluate the efficacy and safety of drugs.Methods:(1)Basic research:All 90 BALB/c female mice were clean class.15 healthy mice were selected as normal group(A group),after the successful modeling(based on the results of preliminary experiment,the seventh day after the model,the detection of mice were accord with successful model requirement,the.success rate was 100%)removed 10 dead mice,the left 65 mice with successful modeling were randomly divided into the control group(B group),low-does group(C group),middle-does group(D group),high-does group(E group)and ciclosporin(Sandimmun)group(F group),13 mice in each group.The mice of C,D and E groups were given Bushen Jianpi Quzhanggxue Granules 0.78g/kg.d,1.56g/kg.d and 3.12g/kg.d respectively.The mice of F group were lavaged with ciclosporin 0.5ml,while the B group was given the same amount of normal sodium.Using medicine when the fifth day after making the model which indicating successful by blood routine examination and marrow pathology.The mice of C,D,E group were lavaged by Traditional Chinese medicine,CsA group was lavaged by CsA,B group was lavaged by equivalence saline,twice a day,continuous use of the drug for 15 days.Tesing Blood RT by orbital vein blood collection on the seventh day after making models and the fifteenth day after using drug.In the end,the fifteenth day after taking durg,the bone marrow were taken for pathological examination.Th1/Th2 balance was evaluated by flow cytometry.Meanwhile,T-bet and GATA-3 mRNA expression were assayed by RT-PCR,and protein expression was assayed by Western blot.Further,IFN-γ,TNF-α,IL-2,IL-4 and IL-10 expression in blood were detected by ELISA.(2)Clinical study:According to the inclusion criteria and exclusion criteria,120 patients with aplastic anemia were randomly divided into the Bushen Jianpi Granule group,the control group,the combination of Chinese medicine and Western medicine,and 40 cases in each group.Results:1.Bushen Jianpi Quzhang Granules improved the general state of the Mice.2.Bushen Jianpi Quzhang Granules and F group enhanced the count of WBC,RBC,HGB and PLT,D group of TCM was superior to C group of TCM and E group of TCM and F group of TCM(P<0.05).3.Using the pathological histology to detecte the bone marrow found that the pathological state of D group of TCM were improvd,other groups had no distinct change.4.Bushen Jianpi Quzhang Granules and F group produced higher GATA-3mRNA,STAT-6mRNA expression and lower T-betmRNA,expression.Besides,D group of TCM was superior to C group of TCM,E groupp of TCM and F group(P<0.05).There were no statistical difference among C group of TCM,E groupp of TCM and F group(P>0.05).5.Bushen Jianpi Quzhang Granules and F group produced higher GATA-3 and STAT-6 protein expression and lower T-bet protein expression.Besides,D group of TCM was superior to C group of TCM,E groupp of TCM and F group(P<0.05).There were no statistical difference among C group of TCM,E groupp of TCM and F group(P>0.05).6.Bushen Jianpi Quzhang Granules and F group produced higher expression of IL-4 and IL-10,lower expression of IFN-γ,TNF-αand IL-2.Also,D groupof TCM was superior to C group of TCM,E groupp of TCM and F group(P<0.05).There was no statistical difference among C group of TCM,E groupp of TCM and F group(P>0.05).7.It f-ound that Chinese herbal medicine and Western medicine combined with the patient in the treatment group after WBC,HGB and PLT,were increased to a certain extent compared with before treatment,after treatment and before treatment were statistically significant(P<0.05),and the therapeutic effect than the control group after treatment effect is more obvious(P<0.05).8.Our results are as follows:Chinese herbal medicine and Western medicine group there were 17 patients in remission occurred,the remission rate reached 42.5%,significantly improve the subjects of 12 people,significantly improve the rate of 30%,subjects had no effect after the treatment of 11,invalid rate reached 27.5%.The total effective rate of treatment was 72.5%;In the western medicine control group,18 patients relieved,and the remission rate reached 45%.4 of the subjects improved obviously,the rate of obvious improvement accounted for 10%.After treatment,there were 18 ineffective patients and 45%ineffective patients.The total effective rate of the control group was 55%;Only 17 patients in traditional Chinese medicine group were relieved,the remission rate was 42.5%,11 subjects were significantly improved,the rate of improvement was 27.5%,and 12patients were ineffective,the effective rate was 30%,and the total effective rate was 70%.All these indicated that the effect of Bushen Jianpi Qu granule combined with western medicine on aplastic anemia is better than that of other groups(P<0.05).9.The degree of bone marrow hyperplasia:comparative analysis of Chinese herbal medicine and Western medicine combined with the treatment group of patients after hyperplasia are low(including reduced and extremely reduced)accounted for 55%of the subjects with a control group reached 82.5%,the pure Chinese medicine group of subjects with low proliferation was 42.5%;the results show that compared favorably to improve the bone marrow hematopoietic function and treatment of Bushen Jianpi Quzhang granule combined with western medicine in the treatment of aplastic anemia after subjects before(P<0.05),and control group after treatment,the effect was compared between the Chinese and Western medicine combined with the treatment effect is better,more obviously(P<0.05).Conclusion:1.Bushen Jianpi Quzhang Granules raised the general state of the mice,and improved the state of blood RT and bone marrow pathology.2.The effect of regulating cytokine expression is associated with IFN-γ/T-bet and IL-4/STAT6 signalling pathways,Th1/Th2 imbalance and T-bet/GATA3 imbalance indicating that the therapeutic effect of Bushen Jianpi Quzhang Granules is complex contact and multiple targets,revealing the mechanism of aplastic anemia on the level of transcription.3.The abnormal change of IFN-γ/T-bet and IL-4/STAT6 signalling pathways and Thl/Th2 imbalance played a key role in The onset of the process of aplastic anemia with abnormal immune.4.Bushen Jianpi Quzhang Granules removed hematopoietic inhibition by reducing Immune hyperfunction through cutting down abnormal activation of IFN-γ/T-bet and raising the expression of IL-4/STAT6 signalling pathways improving Th1/Th2 imbalanceand adjusting the secretion of cytokines.5.By comparison with ciclosporin,Bushen Jianpi Quzhang Granules raise the general state of the mice signally,which point out kidney main bone marrow.6.Bushen Jianpi Quzhang worked as a potent way in aplastic anemia treatment,its effect was associated with drug dose.7.Through the comparative analysis found that before and after the treatment of WBC,HGB,PLT,efficiency of treatment,bone marrow hyperplasia degree index:WBC,HGB,PLT these indicators are increased in different degrees,and the Chinese herbal medicine and Western medicine group after treatment indicators to improve the situation than the other two groups of subjects a lot better;the effective rate of treatment:the total effective rate of the Chinese herbal medicine combined with the western medicine group was higher than the other two groups;the degree of myelodysplastic:the bone marrow hematopoiesis of the patients with the combination of Chinese herbal medicine and Western medicine after treatment is better than before the treatment.It shows that Bushen Jianpi Quzhi granule can accelerate the improvement of aplastic anemia,and its therapeutic effect is outstanding.Although a few patients have symptoms of discomfort,the overall medication is relatively safe and reliable. |
