移植前输注间充质干细胞对脑死亡供鼠移植肾炎症细胞浸润及TNF-α和IL-1β表达的影响

Effect of the Infusion with Mesenchymal Stem Cells Before Transplantation on the Renal Inflammatory Cell Infiltration and the Expression of TNF-α and IL-1β in Brain-dead Donor Rats

目的探讨移植前输注间充质干细胞(MSC)对脑死亡供鼠移植肾炎症细胞浸润及TNF-α和IL-1β表达的影响。方法将健康雄性大鼠随机分为活体供肾移植组、脑死亡供肾移植组、MSC输注组,每组10只大鼠,通过采用抽血和肾脏摘除组织切片,采用全自动生化分析仪、Masson’s Trichrome、TUNEL、蛋白质印迹法和免疫荧光检测3组大鼠的生化指标、炎性浸润、肾上皮细胞凋亡、TNF-α和IL-1β等相关性。结果3组大鼠移植后生命体征(AMP、HR)在加压前、中后差异均无统计学意义(P>0.05),肾血流动力学差异有统计学意义(P<0.05);术后各时间点,脑死亡供肾移植组血清肌酐高于活体供肾移植组(P<0.001),MSC输注组血清肌酐水平低于脑死亡供肾移植组,但高于活体供肾移植(P<0.05);活体供肾移植组肾组织排列整齐,脑死亡肾移植组上皮细胞出现肿胀,中性粒细胞浸润严重,MSC输注组与脑死亡肾移植组相比肾小管水肿减轻,有极少量中性粒细胞炎性浸润;棕色表示凋亡的上皮细胞,活体供肾移植组平均凋亡率19.5%,脑死亡供肾移植组肾上皮细胞平均凋亡最严重(43.0%),与脑死亡供肾移植组相比,MSC输注组上皮细胞有所好转,凋亡率为29.4%;脑死亡供肾移植组大鼠肾组织中TNF-α蛋白表达(14.89±2.17)明显最高于活体供肾移植组(5.13±1.62),TNF-α蛋白在MSC输注组的表达少于脑死亡供肾移植组(7.42±1.08);脑死亡供肾移植组IL-1β阳性表达明显高于活体供肾移植组,MSC输注组IL-1β阳性表达稍多于脑死亡供肾移植组。结论移植前输注MSC具有减少脑死亡大鼠肾炎性浸润状态,抑制肾上皮细胞凋亡,高含量的MSC减少TNF-α和IL-1β表达,减少肾排除现象发生。

Objective To investigate the effect of the infusion with mesenchymal stem cells(MSC)before transplantation on the renal inflammatory cell infiltration and the expression of TNF-αand IL-1βin brain-dead donor rats.Methods The healthy male rats were randomly divided into the living donor kidney transplantation(LDKT)group,brain-dead donor kidney transplantation(BDDKT)group and MSC-infusion group with 10 rats in each group.The blood was extracted and the kidney was removed for the tissue sections.The levels of biochemical indexes,inflammatory infiltration,renal epithelial cell apoptosis,TNF-a and IL-1βin the three groups were detected by the automated biochemical analyzer,Masson’s Trichrome,TUNEL,Western blotting and immunofluorescence to analyze the correlations.Results After transplantation,there were no significant differences in such vital signs as AMP and HR between the three groups before,during and after compression respectively(P>0.05).The renal hemodynamics were statistically different between the three groups(P<0.05).At each time point after surgery,the serum creatinine level of the BDDKT group was significantly higher than that of the LDKT group(P<0.001),and that of the MSC-infusion group was significantly lower than that of the BDDKT group and significantly higher than that of the LDKT group(P<0.05).The renal tissues were well arranged in the LDKT group,the epithelial cells were swollen and the neutrophil infiltration was serious in the BDDKT group,and the renal tubular edema was reduced and there was a small amount of neutrophil inflammatory infiltration in the MSC-infusion group when compared with the BDDKT group.Brown indicated the apoptotic epithelial cells.The average apoptosis rate of renal epithelial cells in the LDKT group was 19.5%,and that in the BDDKT group was 43.0%and the highest among the three groups.Compared with the BDDKT group,the apoptosis of epithelial cells in the MSC-infusion group was improved with the apoptosis rate of 29.4%.The TNF-αprotein expression of the kidney tissue in the BBDKT group(14.89±2.17)was significantly higher than that in the LDKT group(5.13±1.62)and higher than that in the MSC-infusion group(7.42±1.08).The positive expression of IL-1βin the BBDKT group was significantly higher than that in the LDKT group and slightly higher than that in the MSC-infusion group.Conclusion The infusion with MSC before transplantation can improve the nephropathic infiltration state of brain-dead donor rats and inhibit the apoptosis of renal epithelial cells.The high content of MSC reduces the expression of TNF-αand IL-1βand the incidence of renal exclusion.