脂肪间充质干细胞对肾缺血再灌注损伤的保护作用

Protective effect of autologous adipose-derived mesenchymal stem cells on renal ischemia reperfusion injury

目的观察自体脂肪间充质干细胞(adipose-derived mesenchymal stem cells,ADMSCs)对缺血再灌注(ischemia-reperfusion,IR)所致的急性肾损伤的保护及修复作用。方法将成年雄性SD大鼠24只随机分为3组:对照组,模型组,治疗组,每组8只。大鼠在IR建模前14 d,取其大网膜周围的脂肪组织制备自体ADMSCs,并在建模前1d测定大鼠的血肌酐、尿量及尿白蛋白/肌酐比值作为对照。模型组和治疗组制备缺血再灌注肾损伤模型,对照组只打开腹腔。建模1h后,将自体ADMSCs经大鼠尾静脉移植到治疗组,对照组和模型组给予同等体积的生理盐水。细胞移植24 h、72 h后取大鼠血清和尿液检测血肌酐、尿量及尿白蛋白/肌酐比值;在72 h后取肾脏组织通过HE染色进行病理组织学观察;免疫印迹(Western blot)法检测肾脏组织内血红素氧化酶1(heme oxygenase-1,HO-1)及磷酸酰胺腺嘌呤二核苷酸醌氧化还原酶1(NAPDH quinineoxidoreductase-1,NQO-1)的相对表达水平。结果治疗组血清肌酐水平和尿白蛋白/肌酐比值较模型组明显减轻(P<0.05);病理组织学观察显示,模型组大鼠肾脏肾小管存在形态学损伤,而治疗组大鼠肾脏病理损伤明显轻于模型组;Western blot分析显示治疗组抗氧化标志物NQO-1和HO-1的蛋白表达水平显著高于模型组(P<0.05)。结论 ADMSCs经大鼠尾静脉移植后,对IR所致的肾损伤具有明显的修复作用,其修复肾脏损伤机制可能与抗氧化应激有关。

Objective To observe the protective effects of tail veil injection of autologous adi- pose-derived mesenchymal stem cells（ADMSCs） on the renal ischemia reperfusion（IR） injury of rats. Methods Adult male Sprague-Dawley （SD） rats（n = 24） were equally randomized into sham operation group（sham control）, IR injury model group（IR plus saline only）, and cell therapy group（IR plus in- travenous administration of 1.0 × 10^6 autologous ADMSCs）, n = 8 each group. Fourteen days before IR, the adipose tissue around the greater omentum was cut for the preparation of ADMSCs. Twenty- four h before modeling, serum creatinine, urine volume and urinary albumin/creatinine ratio were e- valuated as blank controls. The renal IR injury model was set up. In the sham group, the middle inci- sion was cut and sutured. One h after IR, ADMSCs were transplanted through tail vein of rats in cell therapy group, and the same volume of normal saline was injected into the rats of sham operation group and model group. The renal functional indexes including serum creatinine, urine volume and uri- nary albumin/creatinine ratio were detected by biochemical method 24 h and 72 h after IR. The patho- logical changes were observed by HE staining, and the relative expression of HO-1 and NQO-1 was detected by Western blotting. Results The results of biochemical detection（serum creatinine and uri-nary albumin/creatinine ratio） showed that the kidney function of rats in cell therapy group was signif- icantly better than in model group（P〈0. 05）. The histopathological observation showed that the obvi- ous morphological damage was observed in the renal tubule in model group, and amelioration was ob- served in cell therapy group. Western blotting showed notably higher expression of NAD（P）H qui- none oxidoreductase 1 （NQO1） and HO-1 proteins, two indicators of anti-oxidative capacity, in cell therapy group than in model group. Conclusions ADMSCs therapy minimized kidney damage after IR injury through suppressing oxidative stress.