内皮祖细胞修复缺血再灌注大鼠肾损伤  

Endothelial progenitor cells repair ischemia-reperfusion injury in rats

作　　者：周丽娜[1] 王玉新[1] 方林[2] 武挺[1] 余毅[3] 

机构地区：[1]厦门市第二医院肾内科,福建省厦门市361021 [2]厦门大学附属中山医院泌尿外科,福建省厦门市361004 [3]解放军南京军区福州总医院血液净化室,福建省福州市350025

出　　处：《中国组织工程研究》2014年第32期5146-5151,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:内皮祖细胞在损伤信号的诱导刺激下能被招募到血管损伤局部,分化为成熟内皮细胞,参与血管新生和内皮修复。目的:验证内皮祖细胞能否促进缺血再灌注大鼠肾损伤的修复。方法:抽取大鼠外周血,采用密度梯度离心的方法分离、培养内皮祖细胞。22只雄性SD大鼠随机分为3组,缺血再灌注组采用右肾摘除左肾蒂夹闭左肾动脉、静脉40 min后松开制备缺血再灌注模型;正常对照组除未给予左肾动静脉夹闭外,余操作同缺血再灌注组模型组;内皮祖细胞组动物同样制备缺血再灌注模型,夹闭40 min后从左肾动脉移植内皮祖细胞(5×109 L-1,共1 mL)至左肾,缺血再灌注组予注射同等剂量的细胞培养液。于术后第1天收获3组大鼠肾脏和血标本。流式细胞仪及细胞免疫荧光鉴定内皮祖细胞表面标志(CD34/血管内皮生长因子受体2);测定血尿素氮和肌酐;行免疫组织化学观察各组CD34表达情况。结果与结论:与正常对照组比较,缺血再灌注组大鼠的尿素氮、肌酐均明显升高,肾组织CD34表达减少(P<0.05);给予内皮祖细胞治疗后,缺血再灌注组大鼠的尿素氮、肌酐较治疗前明显下降,肾组织血管内皮细胞标志物CD34的表达较治疗前明显增加(P<0.05)。结果证实内皮祖细胞移植可促进缺血再灌注大鼠肾脏损伤的修复。BACKGROUND： Endothelial progenitor cells are recruited into local vascular injury under the injury-induced stimulation, and then differentiate into mature endothelial cells that are thereby involved in angiogenesis and endothelial repair. OBJECTIVE： To investigate whether endothelial progenitor cells can alleviate renal injury and improve renal function of ischemia/reperfusion injury（I/R） rats. METHODS： Peripheral blood samples extracted from Sprague-Dawley rats were used to isolate and culture endothelial progenitor cells using density gradient centrifugation. Twenty-two male Sprague-Dawley rats were randomized to three groups： I/R group, normal control group, and endothelial progenitor cells group. In the I/R and endothelial progenitor cells groups, the right kidney was removed and the renal artery and vein of the left kidney were occluded for 40 minutes to establish I/R models in the rats, and then endothelial progenitor cells（5×109/L, totally 1 mL） or solvent was transplanted via the artery of the left kidney into the left kidney. In the normal control group, the experimental procedure was same as that in the I/R group except for occlusion of theartery and vein of the left kidney. Renal and blood samples from three groups were collected at day 1 after operation. Peripheral blood CD34 and vascular endoethelial growth factor receptor 2 expressions were determined using flow cytometry and immunofluorescence methods, serum creatinine and urea nitrogen were tested, and immunohistochemistry observation was used for CD34 observation. RESULTS AND CONCLUSION： Compared with the normal control group, serum creatinine and urea nitrogen levels were significantly increased, and tubulointerstitial CD34 expression was decreased in the I/R group（P 〈 0.05）. Endothelial progenitor cells treatment largely decreased the levels of serum creatinine and urea nitrogen, and increased CD34 expression（P 〈 0.05）. These findings indicate that transplantation of endothelial progenitor cells contributes to ren

关 键 词：内皮细胞 再灌注损伤 肾 血尿素氮 干细胞 移植 内皮祖细胞 外周血 缺血再灌注损伤 肾脏 尿素氮 肌酐 CD34 

分 类 号：R394.2[医药卫生—医学遗传学]