重组人促红细胞生成素联合骨髓间充质干细胞移植对大鼠体外循环肾炎症反应的影响  

Effects of recombinant human erythropoietin and bone marrow mesenchymal stem cell transplantation on renal inflammatory response following cardiopulmonary bypass in rats

作　　者：刘凯[1] 李宗杰[1] 吴海卫[1] 王常田[1] 许飚[1] 李德闽[1] 

机构地区：[1]南京大学医学院附属金陵医院(南京军区南京总医院)心胸外科,南京医学硕士研究生210002

出　　处：《医学研究生学报》2016年第4期364-368,共5页Journal of Medical Postgraduates

基　　金：南京军区医学科技创新课题(ZX27)

摘　　要：目的促红细胞生成素(erythropoietin,EPO)及骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)均对肾损伤炎症反应有保护和修复作用,但两者联合应用保护CPB所致肾炎症反应少见报道。文中观察重组人EPO联合BMSCs移植对体外循环(cardiopulmonary bypass,CPB)大鼠肾炎症反应的影响。方法 40只雄性SD大鼠随机数字表法分为5组(n=8):假手术组、CPB组、EPO组、BMSCs组、EPO+BMSCs组,假手术组只建立CPB模型,不行CPB,其余4组均行CPB,流量100 m L/(kg·min)转流并维持1 h,转流后BMSCs组经颈静脉输注BMSCs1.5×106;EPO组颈静脉注射EPO 3000IU/kg;EPO+BMSCs组EPO、BMSCs联合应用;CPB组和假手术组颈静脉输注相同容量的等渗盐水。转流结束后24 h麻醉处死实验动物,检测大鼠血清中肌酐(Cr)、尿素氮(BUN)含量;采用HE染色观察肾组织形态;ELISA法测定血清IL-6、IL-10水平,Western Blot检测肾组织中TNF-α、IGF-1的表达。结果与假手术组比较,CPB组大鼠血清Cr、BUN、IL-6、IL-10和肾组织中TNF-α表达量均升高,而IGF-1表达下降,差异均有统计学意义(P<0.05)。EPO、BMSCs和EPO+BMSCs组血清Cr、BUN水平、IL-6、肾组织中TNF-α均较CPB组下降,而IL-10、肾组织中IGF-1水平较CPB组升高(P<0.05),EPO+BMSCs组血清Cr[(30.26±0.11)μmol/L]、BUN[(6.69±0.20)mmol/L]、IL-6[(401.7±6.8)pg/m L]水平低于EPO组[(42.76±2.24)μmol/L、(9.47±0.50)mmol/L、(533.7±7.0)pg/m L]和BMSCs组[(43.84±3.03)μmol/L、(8.80±0.32)mmol/L、(609.4±6.1)pg/m L],而EPO+BMSCs组IL-10水平[(1055.2±2.8)pg/m L]高于EPO组[(535.1±3.0)pg/m L]和BMSCs组[(544.7±2.6)pg/m L],差异均有统计学意义(P<0.05)。结论 EPO和BMSCs对大鼠CPB肾损伤具有保护作用,效果好于单一使用EPO或BMSCs,减轻CPB对肾的炎症反应可能是其保护机制。Objective The aim of the study was to investigate the effects of recombinant human erythoropoietin（ EPO） and bone marrow mesenchymal stem cell（ BMSC） transplantation on renal inflammatory response following cardiopulmonary bypass（ CPB）.Methods Forty sprague-Dawley male rats were randomly divided into five groups（ n = 8） ： shame operation group,CPB group,EPO group,BMSC group and EPO ＋ BMSC group. CPB model was built in shame operation group without CPB. The other four groups underwent CPB,following by jugular vein infusion of 1. 5 × 106 BMSCs after an hour＇ s 100 L / kg / min bypass. Jugular vein infusion of 3000 IU / kg EPO was done in EPO group,while the combination of EPO with BMSCs was infused in EPO ＋ BMSC group. The same volume of isotonic saline solution was infused via jugular vein in CPB group and shame operation group respectively. Rats were sacrificed at 24 hours after CPB termination. Blood samples were collected for the determination of creatinine（ Cr） and urea nitro（ BUN） levels. HE staining was applied in the examination of renal tissues. ELISA was used in the determination of serum interleukin 6（ IL-6） and interleukin 10（ IL-10） levels and western blot was taken to test the expressions of tumor necrosis factor（ TNF-α） and insulin-like growth factor 1（ IGF-1）. Results In CPB group,the levels of Cr,BUN,IL-6 and the expression of TNF-α were increased,while IL-10 level and of IGF-1 expression were decreased（ P〈 0. 05）. TNF-α expression was increased while IGF-1 expression was decreased in renal tissue（ P〈 0. 05）. HE staining results showed the renal injury in EPO ＋ BMSC group was significantly lower than those in EPO group,BMSC group and CPB group,along with the decrease in the levels of Cr,BUN,IL-6,the increase in IL-10 level（ P〈 0. 05）,as well as the decline of TNF-α expression and the rise of IGF-1 expression（ P 〈0. 05）. Conclusion The combination of EPO and BMSCs which reduces renal inflammatory response following CPB ha

关 键 词：体外循环 肾损伤 骨髓间充质干细胞 重组人促红细胞生成素 炎症因子 

分 类 号：R654.1[医药卫生—外科学]