脂肪源性干细胞输注剂量对抑制大鼠心脏移植急性排斥反应的影响

Effect of Adipose-derived Mesenchymal Stem Cells by Different Doses on the Acute Rejection in Rat Cardiac Transplantation Models

目的观察供体同系脂肪源性干细胞(ADSCs)对大鼠心脏移植急性排斥反应的抑制作用,并探讨其输注剂量对其抑制作用的影响。方法以Wistar、SD大鼠为供受体建立心脏颈部异位移植模型,术毕静脉输注不同剂量ADSCs。实验分组:(1)对照组,注射等体积3ml 0.9%Na Cl;(2)输注ADSCs,0.1×10~6/只组;(3)输注ADSCs,1×10~6/只组;(4)输注ADSCs,10×10~6/只组;每组12只。比较各组移植心脏存活时间和组织病理学改变,及检测血清中IL-2和IL-10的表达。结果对照组移植心脏平均存活6.2±1.2天,输注不同剂量ADSCs(0.1×10~6/只,1×10~6/只,10×10~6/只)各组移植心脏平均存活分别为10.0±1.3天,13.3±2.2天,21.0±4.9天。与对照组比较,输注各组移植心脏存活均得到显著延长(P<0.05)。病理学检查发现ADSCs输注组淋巴细胞浸润、心肌间质水肿、心肌细胞损害均较对照组减轻。与对照组比较,ADSCs输注组大鼠血清中IL-2浓度随着ADSCs浓度增加而逐步降低,且ADSCs 10×10~6/只组差异有统计学意义(P=0.021);IL-10浓度呈剂量相关性逐渐增高,各组间比较差异有统计学意义(P<0.05)。结论静脉输注供体同系ADSCs,可以延长大鼠颈部移植心脏的存活时间,减轻急性排斥反应的程度,并具有一定剂量依懒性;其中以10×10~6/只剂量的抗排斥作用最显著。

Objective To investigate effect of adipose-derived mesenchymal stem cells on the acute rejection in rat cardiac transplantation models and study the possible enhancement role by infusion more ADSCs.Methods Forty-eight SD rats bearing transplanted heart from Wistar rats were divided randomly into 4 groups （n=12）：①Control group：3ml 0.9%NaCl solution was injected through vena after heart transplantation. ②Treatment groups（Ⅰ,Ⅱ,Ⅲ）：0.1×10^6 ADSCs or 1×10^6 ADSCs or 10×10^6 ADSCs in 3ml 0.9%NaCl solution was injected respectively through vena after heart transplantation. One week after transplantation, 6 rats randomly from each group were anaesthetized to get blood and allografts. Grafts pathology was studied. Serum levels of IL-2 and IL-10 was detected by method of ELISA. The rest 6 rats in each group was observed survival time of grafts.Results The survival time of grafts：Control group was 6.2±1.2days, Treatment groupⅠ,Ⅱ,Ⅲ respectively was 10.0±1.3days, 13.3±2.2days, 21.0±4.9days（P〈0.05）.Pathology grade：It showed that lymphocyte infiltration, myocardial interstitial edema and myocardial cell damage were decreased in treatment groups compared with the control group.Concentrations of serum IL-2：It decreased gradually with the increase of ADSCs concentration. And compared with the control group, there was significant difference in ADSCs 10×10^6 treatment group（P〈0.05）. IL-10：It increased gradually with the increase of ADSCs concentration. And there was significant difference each other（P〈0.05）.Conclusion Intravenous infusion of donor ADSCs could prolong the survival time of the rat cardiac allograft and inhibit acute rejection with a dose-dependent lazy. And the dose of 10×10^6/showed that inhibition was the best.