经肾动脉自体骨髓干细胞移植治疗急性肾小管坏死的实验研究

Treatment of acute tubular necrosis by autologous bone marrow stem cells transplantation through renal artery in the rabbits

目的:采用经肾动脉灌注移植兔自体骨髓干细胞,观察自体骨髓干细胞移植对缺血再灌注(I/R)后急性肾小管坏死肾功能的作用及其机制。方法:28只日本大耳白兔采集骨髓,分离自体骨髓干细胞,5-溴-2-脱氧尿嘧啶核苷(5-bromo-2-deoxyuridine,BrdU)标记。全部实验兔建立肾脏I/R后随机分为移植组和对照组。移植组于肾血流恢复后经肾动脉灌注自体骨髓干细胞悬液,对照组同样方法注射等体积生理盐水。分别于I/R前和I/R后第1、3、5、7、14、21、28 d静脉采血,检测血清肌酐(SCr)和尿素氮(BUN),相同时点取肾组织行普通病理及BrdU免疫组化观察。结果:I/R后第1 d和第3 d 2组动物SCr、BUN均达到最高水平,以后逐渐下降;第7 d后移植组SCr及BUN水平逐渐低于对照组,到实验观察结束时的第28 d,对照组SCr和BUN分别为(135.6±32.5)μmol/L和(10.9±2.5)mmol/L,移植组SCr和BUN分别为(90.1±11.1)μmol/L和(8.0±1.5)mmol/L,2组间有显著差异(P<0.05)。普通病理观察,I/R后肾小管上皮细胞变性、坏死甚至脱落,BrdU免疫组化染色显示移植组于细胞移植后第5、7 d肾小管BrdU阳性染色并持续至实验结束,对照组阴性。结论:骨髓干细胞移植可以在一定程度上加速急性肾小管坏死后肾功能修复,自体骨髓干细胞移植是1种加速急性肾小管坏死后修复的安全而有效的方法。

AIM： To observe the effects and location of autologous bone marrow stem cells （BMSCs） transplanted through renal artery into ischemic - reperfusion （I/R） injured kidney. METHODS： BMSCs were collected from rabbits after isolated and then labeled with 5 - bromo - 2 - deoxyuridine （BrdU）. Twenty - eight rabbits were subjected to clamping renal pedicles for 105 rain and divided into the transplantation group and control group randomly. BrdU labeled BMSCs or saline were injected into the kidney by renal artery, respectively. Before and after I/R at the 1st, 3rd, 5th, 7th, 14th, 21th and 28th d, the venous blood was collected to measure serum Cr and BUN. In the same time, renal tissue was collected for pathological and immunohistochemical study. RESULTS： After I/R, serum Cr and BUN levels in the rabbits in two groups became higher, and on the 1st and 3rd d after I/R, reached the highest level. On the 7th d the serum Cr and BUN levels in transplantation group were lower than those in control group. On the 28th d the levels of serum Cr （90. 1 ± 11.1 ） μmol/L and BUN （ 8.0 ±1.5 ） mmol/L in transplantation group were significantly lower than those in control group （ 135.6 ±32. 5） μmol/L and （ 10.9± 2. 5 ） mmol/L, respectively （ P 〈 0. 05 ）. Pathological observation of renal tissue showed the degeneration, necrosis and abscission in renal tubular epithelial cells. The BrdU positive staining by immuno- histochemical study was found in renal tubular in transplantation group on the 5th and 7th d after I/R, and maintained to the end of experiment, but no detection in control group. CONCLUSION： BMSCs transplantation through renal artery accelerates the repairment of renal functions after acute tubular necrosis by ischemic - reperfusion. Autologous BMSCs transplantation is a safe and valid method to accelerate the repairment after acute tubular necrosis.