摘要目的 探索异基因造血干细胞移植联合内皮祖细胞(EPC)输注对骨髓炎性损伤的影响.方法 6～8周龄BALB/c(H-2Kd)小鼠经致死剂量全身照射(TBI)预处理后建立异基因骨髓移植模型(BMT组),并在BMT基础上同时输注体外培养的EPC,建立骨髓联合EPC移植模型(EPC组).移植后第7、14、21、28天分别获取各组小鼠骨髓细胞,流式细胞术检测EPC培养比率及两组嵌合率变化,通过骨髓细胞和外周血细胞计数比较两组骨髓造血情况,Western blot法检测骨髓巨噬细胞表面特异性标志CD68表达水平;实时定量PCR及Western blot法分析骨髓炎性复合体nlrp1、nlrp6、caspase-1mRNA及蛋白表达水平.结果 移植后第7天BMT组嵌合率为(83.69±1.26)％,明显低于EPC组的(91.65±2.77)％(P＜0.05).在移植后第10、15、20天,EPC组小鼠外周血中的WBC、网织红细胞、PLT均高于BMT组(P值均＜0.05),在第7、21天EPC组骨髓中原始红细胞较BMT组高(P值均＜0.05).HE染色显示,移植后第7、14、21天,EPC组骨髓微环境恢复较BMT组快.Western blot检测显示EPC组骨髓CD68蛋白表达水平较BMT组低.EPC组nlrp1、nlrp6、casepase-1 mRNA及蛋白表达水平在移植后各时间点接近正常表达水平,与BMT组相比,差异均有统计学意义(P值均＜0.05).结论 异基因造血干细胞移植联合输注EPC可减少炎性细胞的浸润及减少炎性复合体的活化,促进骨髓造血重建.

abstractsObjective To explore effects of allogeneic hematopoietic stem cell transplantation (HSCT) in combination with infusion of endothelial progenitor cells (EPC) on bone marrow inflammatory injury.Methods 6-8 weeks BALB/c (H-2Kd) mice after lethal dose of irradiation (TBI) were subjected to allogeneic bone marrow transplantation (BMT group) or co-transplantation of EPC (EPC group).Samples of bone marrow cells of mice in each group on days 7,14,21,28 after transplantation were obtained to detect EPC cultural and cell chimeric rates by flow cytometer.Mice were sacrificed on days 7,14,21 and 28 post HSCT to analyze bone marrow pathology by H&E staining,the infiltration of macrophages and neutrophils by Western blot,validation expression levels of inflammatory complexes nlrp1、nlrp6 and its downstream molecules casepase-1 by Q-PCR and Western blot.Results Cell chimeric rate on day 7 after transplantation in EPC group [(91.65 ± 2.77) ％] was significantly higher than in BMT group [(83.69 ± 1.26)％].Alleviated osteomyelitis injury and inflammatory cell infiltration in EPC group were observed when compared with BMT mice.Also significant reductions of the levels of nlrpl、nlrp6、casepase-1 transcription complexes in EPC mice were noted when compared with BMT ones.Conclusion Co-transplantation of HSC and EPC could alleviate inflammatory cell infiltration and activation of the complex to promote the repair of bone marrow.