摘要
目的探讨红细胞生成素(EPO)对慢性肾衰竭(CRF)大鼠外周血内皮祖细胞(EPC)的动员和肾小管周围毛细血管修复的影响。方法采用分阶段5/6肾切除制备大鼠CRF模型。成年雄性6周龄大鼠随机分为3组:假手术组、CRF模型组、EPO组,每组7只。从第3周开始,EPO组大鼠每次皮下注射重组人EPO 50U/kg,每周3次,共6周。8周时处死大鼠并取其外周血分离与培养EPC,并检测EPC的功能。取肾组织,采用逆转录聚合酶链反应(RT-PCR)和免疫组化方法检测肾组织血管内皮细胞生长因子(VEGF)mRNA和蛋白的表达。并通过免疫组化方法检测肾间质CD31的表达来计算肾小管周围毛细血管密度。结果与模型组比较,EPO组外周血EPC动员能力增强[(8.34±0.85)比(4.63±0.64)细胞/视野,P<0.05];外周血EPC增殖能力(MTT值)增强(0.49±0.08比0.18±0.04,P<0.05);外周血EPC黏附能力提高[(27.33±5.61)比(14.83±3.55)细胞/视野,P<0.05];外周血EPC形成血管腔样结构的能力提高[(18.44±3.75)比(9.81±2.23)血管腔/视野,P<0.05]。与模型组比较,EPO组肾间质小管周围毛细血管密度增加;肾组织VEGF mRNA及蛋白的表达增强(均P<0.05)。结论 EPO可动员CRF大鼠外周血EPC,并能促进肾小管周围毛细血管的修复。
Objective To investigate the effects of erythropoietin（EPO）on mobilization of peripheral endothelial progenitor cells（EPC）and repairment of peritubular capillary in rats with chronic renal failure（CRF）. Methods The rat CRF model was established by a two stage 5/6nephrectomy procedure. Rats were randomly divided into three groups（n=7/group）：sham operation,CRF model only,and CRF model＋EPO treatment group. Three weeks after the second stage of nephrectomy,rats in CRF model＋EPO treatment group received subcutaneous injection of human recombinant EPO 50U/kg each time and three times a week for 6weeks,then all rats in three groups were sacrificed. EPC were isolated and cultured from peripheral blood. The ability of cell mobilization,proliferation,adhesion and vasculogenesis in vitro was observed. The protein and mRNA expression levels of vascular endothelial growth factor（VEGF）in kidney were detected by reverse transcription polymerase chain reaction（RT-PCR）and immunohistochemistry method. Renal interstitium CD31 was examined by immunohistochemistry and the density of peritubular capillarycal was calculated. Results Compared with CRF model group,the ability of cell mobilization,proliferation,adhesion and vasculogenesis was promoted by administration of EPO in treatment group[mobilization：（8.34±0.85）vs（4.63±0.64）cells/HP,proliferation（MTT）：0.49±0.08 vs 0.18±0.04,adhesion：（27.33±5.61）vs（14.83±3.55）cells/HP,vasculogenesis：（18.44±3.75）vs（9.81±2.23）/HP,all P〈0.05]. Similarly the density of peritubular capillary in treatment group was increased（P0.05）,and the protein and mRNA expression levels of VEGF up-regulated（all P〈0.05）. Conclusion EPO could mobilize peripheral EPC and promote the repairment of peritubular capillary in CRF rat.
出处
《中华高血压杂志》
CAS
CSCD
北大核心
2014年第12期1167-1173,共7页
Chinese Journal of Hypertension
基金
福建省自然科学基金资助项目(2012J01343)
