rhEPO对大鼠肾缺血再灌注损伤TLR4/NF-κB通路的影响  

Recombinant human erythropoietin attenuates renal ischemia reperfusion injury in rats through inhibiting TLR4 /NF-κB signal transduction pathway

作　　者：李清初[1] 陈超[1] 尹友生[1] 韦家智[1] 李康慧[1] 覃源[1] 

机构地区：[1]桂林医学院附属医院肾内科,广西桂林541001

出　　处：《第三军医大学学报》2014年第17期1817-1821,共5页Journal of Third Military Medical University

基　　金：广西壮族自治区卫生厅课题(Z2008293)~~

摘　　要：目的探讨不同时段相同剂量重组人促红细胞生成素(recombinant human erythropoietin,rhEPO)对大鼠肾缺血再灌注损伤TLR4/NF-κB信号通路的影响。方法将70只雄性SD大鼠按随机数字表法分假手术组,缺血再灌注组(IR),注射rhEPO在缺血前6 h组(P6)、缺血前2 h组(P2)、缺血前0 h组(P0)、缺血后2 h组(A2)、缺血后6 h组(A6),每组10只。测定血尿素氮(BUN)、肌酐(Cr)水平,HE染色观察肾组织病理学形态,免疫组织化学法检测肾组织TLR4、NF-κB p50蛋白表达与分布,RT-PCR法检测肾组织TLR4、NF-κB mRNA表达。结果 IR组的血清BUN、Cr,肾小管Paller评分,TLR4、NF-κB p50蛋白表达及肾组织TLR4、NF-κB mRNA表达较假手术组均升高(P<0.01);P6、P2、P0、A2、A6组的Paller评分、BUN和Cr的血清浓度、TLR4及NF-κB的蛋白质和mRNA表达较IR组减少(P<0.01),每组的5项监测指标趋势一致;rhEPO干预组中P6、P2组的5项监测指标均较P0、A2、A6组减少(P<0.01),P6和P2间及P0、A2和A6组间的各项指标比较差异无统计学意义(P>0.05)。结论 rhEPO干预能缓解肾缺血再灌注损伤所致的急性肾损伤,可能是通过抑制TLR4/NF-κB通路起作用,rhEPO在肾缺血前注射比肾缺血后注射对肾脏保护作用更明显。Objective To determine the effect of recombinant human erythropoietin （rhEPO） injected at the same dose but different time points on toll-like receptor 4/nuclear factor-kappa B （TLR4/NF- κB） signal transduction pathway in rats after renal ischemia reperfusion injury. Methods Totally 70 male SD rats were randomly divided into 7 groups （n = 10） , sham operation group （S group）, ischemia reperfusion group （IR group） , the group receiving rhEPO in 6 h before ischemia （1）6 group, 2 000 U/kg rhEPO via tail vein）, the group receiving rhEPO in 2 h before isehemia （ P2 group）, the group receiving rhEPO in 0 h before ischemia （P0 group） , the group receiving rhEPO in 2 h after ischemia （A2 group）, and the group receiving rhEPO in 6 h after isehemia （A6 group）. The IR injured model was established by resecting the right kidney and blocking left renal artery for 45 min. In 24 h after blocking, all animals were killed and their blood samples and left kidneys were collected. The renal pathological changes were observed by HE staining. Serum levels of BUN and Cr were determined. The expression and distribution of TLR4 and NF-κB p50 in renal tissue were determined by immunohistochemistry assay. Reverse transcriptional-polymerase chain reaction （RT-PCR） was performed to detect the mRNA levels of TLR4 and NF-κB. Results Serum levels of BUN （40.63 ± 4.45 ） and Cr （166.25 ±7.80）, the paller scores of renal tubule pathologic damage （251.83 ± 12.65 ）, the protein expression of TLR4 and NF-κB p50 （211. 758 ± 17. 720 and 212. 492 ± 17. 989）, and mRNA expression of TLR4 and NF-κB （2. 191 ± 0. 046 and 0. 771 ＋±0. 069） were significantly higher in IR group than S group （P 〈 0.01 ）. While, these indexes were obviously lower in the P6, P2, P0, A2 and A6 groups than the IR group （P 〈0.01 ）. Moreover, the changing trend of the 5 test items was accordant in each group. Among theadministrated groups by rhEPO, the 5 test items were markedly lower in P6 and

关 键 词：肾脏 缺血再灌注损伤 重组人促红细胞生成素 TOLL样受体4 NF-ΚB 

分 类 号：R364.12[医药卫生—病理学] R692.02[医药卫生—基础医学]