摘要：
Objective To investigate the role of bone marrow mesenchymal stem cells (BMSCs) pretreated with erythropoietin (EPO) in the prevention of acute rejection after renal transplantation in rats. Methods BMSCs were divided into five groups: control group (without EPO), group A (pretreated with EPO at a final concentration of 10 IU/mL), group B (pretreated with EPO at a final concentration of 100 IU/mL), group C (pretreated with EPO at a final concentration of 500 IU/mL) and group D (pretreated with EPO at a final concentration of 1 000 IU/mL). In each group, the BMSCs were cultured for 24 h and 48 h. The proliferation rate of the BMSCs was determined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. The BMSCs were divided into two groups: BMSC group (without EPO) and EPO-BMSC group (pretreated with EPO at a final concentration of 500 IU/mL). After 48 h culture, Western blot was adopted to measure the expression level of CXC chemokine receptor (CXCR) 4 protein in BMSCs. Wistar rats were used as the donors, and SD rats were utilized as the recipients to establish the rat models with acute rejection after renal transplantation. The recipient rats were randomly divided into four groups (n=6 in each group) including the control group (without any intervention), EPO group (injection of 1 mL of solution containing 500 IU EPO via tail vein immediately after surgery), BMSC group (injection of 1 mL of solution containing 1×106/mL BMSCs via tail vein immediately after surgery) and EPO-BMSC group (injection of 1 mL of solution containing 1×106/mL BMSCs cultured in vitro with 500 IU/mL EPO via tail vein). The level of serum creatinine (Scr) level was determined by Scr detection kit. Western blot was used to detect the expression levels of interferon (IFN)-γ and interleukin (IL)-4 proteins. Results After 24 h culture, the proliferation rate of BMSCs did not significantly differ among all groups (all P>0.05). After 48 h culture, the proliferation rate of BMSCs in group C (pretreated with EPO at a final concentration of 500 IU/mL) was significantly higher than that in the control group (P0.05). At 5 d after operation, the levels of Scr in the EPO, BMSC and EPO-BMSC groups were significantly lower than that in the control group (all P0.05). At 1 d after surgery, compared with control group, the expression levels of IFN-γ protein and IFN-γ/IL-4 ratio in the renal tissues in the EPO, BMSC and EPO-BMSC groups were significantly decreased to varying extents (all P0.05).培养48 h,与对照组比较,C组(EPO终浓度500 IU/mL)的BMSC增殖率较高,差异有统计学意义(P0.05);术后5 d,与对照组比较,EPO组、BMSC组和EPO-BMSC组受体鼠的Scr水平均明显降低,差异均有统计学意义(均为P0.05).术后1 d,与对照组比较,EPO组、BMSC组及EPO-BMSC组受体鼠移植肾组织中IFN-γ蛋白表达水平和IFN-γ/IL-4均有不同程度降低,差异均有统计学意义(均为P<0.05).术后5 d,与对照组比较,EPO组、BMSC组和EPO-BMSC组中IFN-γ蛋白表达水平和IFN-γ/IL-4均有不同程度降低,差异均有统计学意义(均为P<0.05),其中EPO-BMSC组中IFN-γ蛋白表达水平和IFN-γ/IL-4均低于EPO组和BMSC组,差异均有统计学意义(均为P<0.05).结论 EPO预处理BMSC可预防肾移植术后急性排斥反应,保护移植肾功能.
