rHuEPO对盲肠结扎穿孔术+检验性失血大鼠急性肾损伤的保护作用

Protective effects of recombinant human erythropoietin in rats with acute kidney injury under cecal ligation and puncture and blood loss during laboratory test

目的探讨重组人促红细胞生成素(r Hu EPO)对盲肠结扎穿孔术(CLP)+检验性失血大鼠急性肾损伤的保护作用。方法 160只SD大鼠CLP术后按照随机数表法取均分为四组(对照组、小剂量组、中剂量组和大剂量组),每组各40只,四组均关腹后0 h、24 h、48 h尾静脉放血1 ml,小剂量组0 h皮下注射3 000 U/kg+生理盐水1 ml(24 h、48 h皮下注射生理盐水1 ml),中剂量组0 h、24 h皮下注射r Hu EPO 3 000 U/kg+生理盐水1 ml(48 h皮下注射生理盐水1 ml),大剂量组0 h、24 h、48 h皮下注射3 000 U/kg+生理盐水1 ml,对照组0 h、24 h、48 h皮下注射生理盐水1 ml,分笼饲养,72 h每组各取5只经心脏取血2 ml,检测血红蛋白(Hb)、尿素氮(BUN)、肌酐(Cr)、肿瘤坏死因子(TNF-α),检测肾脏丙二醛(MDA)、髓过氧化物酶活性(MPO)、核转录因子(NF-κB)表达和末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸缺口标记(TUNEL)阳性细胞数。结果大中剂量组BUN、Cr、TNF-α、MDA、MPO、NF-κB、TUNEL阳性细胞数分别与对照组比较差异均有统计学意义(P<0.05),大剂量组较中剂量组差异有统计学意义(P<0.05),而小剂量组上述指标分别与对照组比较差异则均无统计学意义(P>0.05),四组的Hb比较差异无统计学意义(P>0.05)。结论 r Hu EPO能够降低盲肠结扎穿孔术+检验性失血所致急性肾损伤大鼠TNF-α、MDA、MPO、NF-κB表达和TUNEL阳性细胞数,对脓毒症和检验性失血双重打击急性肾功能损伤具有一定的保护作用,且呈剂量依赖性。

Objective To discuss the protective effects of recombinant human erythropoietin （rHuEPO） in rats with acute kidney injury under cecal ligation and puncture （CLP） and blood loss during laboratory test. Methods One hundred and sixty SD rats were averaged into four groups （control group, low-dose rHuEPO group, me-dian-dose rHuEPO group and high-dose rHuEPO group） through the random number table after CLP. 1 ml of tail vein blood each was taken 0 h, 24 h, 48 h after the abdomen closure. rHuEPO 3 000 U/kg＋saline 1 ml was injected subcuta-neously in low-dose rHuEPO group at the time of 0 h , and 1 ml of saline was injected respectively at 24 h and 48 h. rHuEPO 3 000 U/kg＋saline 1 ml was injected subcutaneously in median-dose rHuEPO group at the time of 0 h and 24 h, and 1 ml of saline was injected at 48 h. rHuEPO 3 000 U/kg＋saline 1 ml was injected subcutaneously in high-dose rHuEPO group at the time of 0 h, 24 h and 48 h. The control group was injected with normal saline 1 ml each at the time of 0 h, 24 h and 48 h. Rats were separated in different cages for 72 h, then 5 out of each group were selected and drew 2 ml of blood each from the hearts to test hemoglobin hemoglobin （Hb）, urea nitrogen （BUN）, creatinine （Cr）, tu-mor necrosis factorα（TNF-α）, as well as the expression of malonaldehyde （MDA）, myeloperoxidase （MPO） and nu-clear factor-κB （NF-κB） in their kidneys, and also, the amount of terminal dexynucleotidyl transferase （TdT）-mediat-ed dUTP nick end labeling （TUNEL） positive cells. Results The median-dose rHuEPO and high-dose rHuEPO group showed statistically significant differences respectively with the control group in the test results of BUN, Cr, TNF-α, MDA, MPO, NF-κB and TUNEL positive cells （P〈0.05）. The differences in the above indexes between the high-dose rHuEPO group and the median-dose rHuEPO group were statistically significant （P〈0.05）, while the differ-ences between the low-dose rHuEPO group and the control group was not different （P〉0.05）. The four groups showed no statistically significant difference from each other inHb （P〉0.05）. Conclusion rHuEPO could decrease the expres-sion of MDA, MPO, NF-κB and the TUNEL positive cells in the rats with acute kidney injury under the circumstanc-es of CLP and blood loss during laboratory test. It has a certain extent of protective effects for the acute kidney injury caused by sepsis and blood loss during laboratory test, which also demonstrates a feature of dose-dependence.