脐血源EPC联合造血干细胞移植在造血重建和GVHD中作用研究

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针对脐血源内皮祖细胞(endothelial progenitor cell,EPC)与造血干细胞(hematopoietic stem cell,HSC)联合移植方式进行研究,并探讨移植后的造血细胞重建及移植物抗宿主病(graft versus host disease,GVHD)的影响。构建人-鼠移植EPC与HSC模型,采用移植HSC的小鼠为参考对比,分析移植后的体内植入及造血重建状况,并分析小鼠有无GVHD相关症状,最后采用病理对GVHD进行实验验证。EPC与HSC联合移植小鼠组(n=10)的实验组,生存的平均时间约54 d,长时间生存率接近70%;单纯移植小鼠组(n=10)的实验组中,生存的平均时间约51 d,长时间生存率达到60%;EPC与HSC联合移植小鼠组与单纯移植小鼠组进行比较,生存时间有显著的增高趋势,但经统计学分析两者比较无显著差异性(p=0.314)。在血细胞恢复实验中,联合移植组相比较单纯组,随着时间的增加,HBG、WBC及PLT的恢复值均有增高趋势。联合移植组小鼠骨髓中人CD45比例为(18.21±10.57)%,单纯组骨髓中人CD45嵌合比例为(13.27±6.36)%,无明显统计学差异性(p=0.308);在联合移植组脾脏中的嵌合比例为(5.14±2.97)%,在单纯移植组比为(4.91±7.56)%,无显著统计学差异(p=0.840)。在联合移植组中检查到2只小鼠带有GVHD病症,表现是轻中度活动度明显减少和体重有下降趋势;单纯移植组检查到3只小鼠带有GVHD病症,表现为活动度下降、体重下降、弓背姿势及耸毛现象;共同点为带有GVHD症状的小鼠均有骨髓与小肠不同损伤情况。本研究初步认为脐血源内皮祖细胞与造血干细胞联合移植的方式,能够对移植后的造血系统重建及移植物抗宿主病产生一定的积极影响,但是效果并不显著。 To investigate the combined transplantation of endothelial progenitor cells (EPCs) and hematopoietic stem cells (HSC) in umbilical cord blood and to investigate the relationship between hematopoietic cell reconstitution and graft versus host disease GVHD) effects. The human and mouse transplanted EPC and HSC models were constructed. The transplanted HSCs were used as a reference to compare the in vivo implantation and hematopoietic reconstitution after transplantation, and to analyze whether the mice had GVHD-related symptoms. Finally, pathological examination of GVHD was performed . In the experimental group co-transplanted with EPC and HSC (n = 10), the mean survival time was about 54 days, and the long-term survival rate was close to 70%. In the experimental group of pure transplanted mice (n = 10) The average time was about 51 days, and the long-term survival rate was 60%. The survival time of EPC and HSC transplantation group was significantly higher than that of pure transplantation group, but there was no significant difference between the two groups Sex (p = 0.314). In the blood cell recovery experiment, the recovery values ​​of HBG, WBC and PLT tended to increase with the increase of time in the combined transplantation group compared with the simple group. The ratio of human CD45 in the bone marrow of the combined transplantation group was (18.21 ± 10.57)%, while that of the human bone marrow group in the simple group was (13.27 ± 6.36)%, with no significant difference (p = 0.308) The percentage of chimerism in the spleen was (5.14 ± 2.97)% in the spleen group and (4.91 ± 7.56)% in the simple graft group, with no significant difference (p = 0.840). Two mice with GVHD were detected in the combined transplantation group with a significant reduction in mild to moderate activity and a tendency to decrease in body weight; three mice with GVHD were found to have a decreased activity in the transplant group , Weight loss, arched posture and hair-raising phenomenon; the common point is that mice with GVHD symptoms have different injuries of bone marrow and small intestine. This study suggests that the combined transplantation of umbilical cord blood-derived endothelial progenitor cells and hematopoietic stem cells may have some positive effects on the reconstruction of hematopoietic system and graft-versus-host disease after transplantation, but the effect is not significant.
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