间充质干细胞和内皮祖细胞联合输注对造血干细胞移植后损伤肺的影响

Effects of Combined Infusion of Mesenchymal Stem Cells and Endothelial Progenitor Cells on Lung Injury after Hematopoietic Stem Cell Transplantation

目的:探索骨髓间充质干细胞(mesenchymal stem cell,MSC)和内皮祖细胞(endothelial progenitor cell,EPC)联合输注对造血干细胞移植(hematopoietic stem cell transplantation,HSCT)后损伤肺的影响。方法:实验分为正常对照(normal)、单纯照射(TBI)、骨髓细胞移植(BMT)、骨髓移植联合EPC输注(BMT+EPC)、骨髓移植联合MSC输注(BMT+MSC)、骨髓移植联合EPC和MSC输注(BMT+EPC+MSC)共6组,建立HSCT模型,于移植后d 30脱颈处死,取肺组织,待测。RT-PCR检测VEGF,IL-18,IL-12b的mRNA表达水平,Western blot法检测NLRP3蛋白表达水平,HE染色观察肺组织病理学形态,免疫组织化学观察MPO,CD146+细胞表达情况。结果:VEGF基因在BMT+EPC+MSC组的表达水平明显高于其他各组(P<0.01),而BMT+EPC、BMT+MSC和BMT组基因表达水平相近,且均高于正常对照组(P<0.05)。IL-18,IL-12b基因的表达水平在BMT组最高,BMT+EPC+MSC组最低,与对照组之间比较差异有统计学意义(P<0.05)。HSCT能引起NLRP3蛋白表达增高,BMT+EPC+MSC组可明显降低肺细胞NLRP3蛋白水平,趋于正常。与正常组织相比,BMT+EPC+MSC组能更有效地改善肺组织结构,MPO+细胞表达低,CD146+细胞表达高。结论:联合输注MSC和EPC能促进HSCT后损伤肺的毛细血管新生,减轻炎症反应,促进肺的修复,优于单一的治疗方法。

Objective:To investigate the effects of combined infusion of mesenchymal stem cells(MSC)and endothelial progenitor cells(EPC)on lung injury after hematopoietic stem cell transplantation(HSCT).Methods:The experiment was divided into normal control group,irradiation group,bone marrow cell transplantation group(BMT group),BMT+EPC group,BMT+MSC group and BMT+EPC+MSC group.The model of HSCT was established,on the 30th day after transplantation,the mice were sacrificed.Then lung tissue was taken for testing.The mRNA expression levels of VEGF,IL-18,IL-12b were detected by RT-PCR,and protein expression level of NLRP3 was detected by Western blot.The expression of MPO and CD146 was observed by immunohistochemistry assay.Results:The expression level of VEGF gene in BMT+EPC+MSC group was significantly higher than that in other groups(P<0.01).The expression level of IL-18 and IL-12b gene was the highest in BMT group and the lowest in BMT+EPC+MSC group,and the difference was statistically significant(P<0.05).HSCT could increase the expression of NLRP3 protein,and the BMT+EPC+MSC could significantly reduce the level of NLRP3 protein in lung cells,tending to normal.Compared with normal tissues,the BMT+EPC+MSC could improve the lung tissue structure more effectively,the expression of MPO positive cells was lower,and the expression of VEGF positive cells was higher.Conclusions:The combined infusion of MSC and EPC can promote capillary regeneration,alleviate inflammation and promote lung repair after HSCT,which is superior to single EPC or MSC infusion.