摘要:
Objective To explore the effect of faeces bombycis extract combined with cyclosporine A treatment on proliferation of hematopoietic progenitor cells in aplastic anemia(AA)rat model as well as their influence on the differentiation of myeloid PU.1 and erythroid differentiation of JAK2/STAT5 signaling pathway.Methods The Wistar rats were randomly divided into five groups,model group,faeces bombycis extract group,cyclosporine A (CsA)group,combination group and normal group.Hematopoietic stem cells of rats were extracted in vitro and were directionally induced to differentiate into hematopoietic progenitor cells of granulocytes and erythrocytes.The colony forming units(CFU)formed by hematopoietic progenitor cells of each group were counted.Total mRNA and protein were extracted and the levels of PU.1 mRNA and protein were detected with real-time polymerase chain reaction(qPCR)and Western blotting.Erythroid cell mRNA was extracted and the levels of JAK2,STAT5,GATA1 mRNA were detected with qPCR.Results ① Compared with the model group,assay of colony forming unit-granulocyte macrophage(CFU-GM),assay of colony forming unit-erythrocyte(CFU-E)and burst forming unit-erythrocyte (BFU-E)in each treatment group were increased (all P< 0.05),and CFU-E,BFU-E and CFU-GM in the combination group were significantly increased(all P<0.05).Compared with the model group,PU.1 mRNA and protein levels of granulocytes in each treatment group were increased in a varying degree,while those in the combination group were increased significantly as compared with the other treatment groups(all P<0.05).③ Compared with the model group,JAK2,STAT5 and GATA1 mRNA levels of erythroid cells in each treatment group were increased;and those in the combination group present a remarkable growth compared with the other treatment groups.Conclusion Faeces bombycis extract may promote bone marrow function of AA rat model through increasing the expression of PU.1,JAK2,STAT5 and GATA1 while faeces bombycis extract and cyclosporine A have a synergistic effect on bone marrow function of AA rat model.%目的 探讨蚕砂提取物联合环孢菌素A(Cyclosporine A,CsA)对再生障碍性贫血(aplastic anemia,AA)大鼠模型造血祖细胞增殖的影响,及对粒系增殖分化PU.1和红系增殖分化JAK2/STAT5信号通路的影响.方法 将Wistar大鼠随机分为正常组、模型组、蚕砂组、CsA组、联合组,共5组.体外提取各组大鼠造血干细胞,定向诱导其向粒单核系、红系分化,对各组造血祖细胞集落形成单位(colony forming units,CFU)计数.提取粒单核系祖细胞总RNA和蛋白,采用实时定量PCR(real-time PCR,qPCR)和Western blot技术检测PU.1 mRNA及蛋白表达情况,提取红系祖细胞总RNA,采用qPCR技术检测细胞JAK2、STAT5、GATA1 mRNA表达情况.结果 ①各治疗组红系造血祖细胞集落形成单位(colony forming unit-erythrocyte,CFU-E)、红系爆式集落形成单位(burst forming unit-erythrocyte,BFU-E)和粒单核系造血祖细胞集落形成单位(colony forming unit-granulocyte macrophage,CFU-GM)数目较模型组均有不同程度增高,差异有统计学意义(均P<0.05);其中联合组CFU-E、BFU-E、CFU-GM升高明显(均P<0.05).②各治疗组粒单核系细胞PU.1 mRNA和蛋白含量较模型组均有不同程度升高,其中联合组升高明显(均P<0.05).③各治疗组红系细胞JAK2、STAT5、GATA1 mRNA表达较模型组有不同程度升高,联合组表达高于其他各干预组.结论 蚕砂提取物可能通过增加PU.1 mRNA和蛋白表达及JAK2、STAT5、GATA1 mRNA表达促进AA大鼠骨髓造血作用,且蚕砂提取物联合CsA对AA大鼠骨髓造血具有协同增效作用.