摘要
目的 建立大鼠孤立肾保留肾单位手术（nephron-sparing surgery,NSS）动物模型,探讨脂肪来源的基质血管成分（stromal vascular fraction,SVF）对NSS急性肾损伤（acute kidney injury,AKI）的修复作用.方法 2014年10月至2015年2月,36只SD大鼠行右肾切除术,建立孤立肾动物模型,并采用随机数表法分为3组：①实验组,夹闭左侧肾动脉,阻断肾血流45 min,行左侧NSS,构建左侧孤立肾AKI模型.将预先制备的自体脂肪SVF肾组织内注射.②对照组,构建上述NSS后AKI模型,仅采用等量的PBS注射.③假手术组,充分游离左肾,不行NSS.于术后24、72 h分别收集血标本,检测血清肌酐（SCr）、尿素氮（BUN）水平;获取肾组织行HE、免疫组化检测,评价肾小管损伤程度和细胞增殖、凋亡及管周毛细血管新生特征.结果 NSS后24、72 h,实验组SCr分别为（95.30±4.48）、（60.72±8.30） μmol/L,低于对照组的（143.77±4.34）、（157.50±8.27） μmol/L,差异有统计学意义（P＜0.05）;实验组BUN分别为（21.90±1.96）、（15.50±1.45） mmol/L,低于对照组的（29.47±1.17）、（33.20±4.13） mmol/L,差异有统计学意义（P＜0.05）.实验组NSS后24、72 h肾小管损伤评分为1.8±0.5、2.0±0.5,亦低于对照组的2.7±0.5、3.8±0.6,差异有统计学意义（P＜0.05）;PCNA阳性细胞数为66.2±4.1、78.7±5.5,高于对照组的30.0±4.8、54.3±3.9,差异有统计学意义（P＜0.05）;凋亡细胞数为28.4±3.3、36.3±3.1,低于对照组的54.5 ±4.1、87.4±3.0,差异有统计学意义（P＜0.05）;实验组CD34阳性管周毛细血管数为98.3±11.0、135.8±13.0,高于对照组的54.4±8.4、44.0±6.1,差异有统计学意义（P＜0.05）,与假手术组85.5±8.1、114.9±10.3相比差异无统计学意义（P＞0.05）.结论 SVF可促进大鼠NSS相关性AKI的结构和功能恢复,可为NSS相关性AKI的防治提供一种新策略,有待进一步研究.
Objective To study the protective effect of adipose derived stromal vascular fraction （SVF） against acute kidney injury （AKI） after nephron-sparing surgery （NSS） in rats.Methods Left solitary kidney （SK） model was developed by right kidney nephrectomy in SD rats,which were divided into 3 groups randomly：experiment group （NSS ＋ SVF/PBS）,control group （NSS ＋ PBS）,and sham group.During NSS,renal artery was clamped for 45 minutes to induce AKI,after which SVF/PBS was administrated through intra-parenchymal injection in experimental group,while PBS was injected in control group.After 24 h and 72 h,blood sample was collected for the evaluation of serum creatinine （SCr） and blood urea nitrogen （BUN）.In addition,kidney tissues were retrieved for hematoxylin and eosin （HE） staining and immunohistochemical staining.Kidney injury score as well as characteristics of cells proliferation,apoptosis and angiogenesis of peri-tubular were also evaluated.Results At 24 h and 72 h after NSS,serum levels of SCr and BUN in experimental group were significantly lower than that in control group,but were still higher than that in sham group （P 〈 0.05）.HE staining indicated that injury score in experimental group was significantly lower than that in control group at 24h and 72h （1.8 ± 0.5 versus 2.7 ± 0.5,and 2.0 ± 0.5 versus 3.8 ± 0.6,P 〈 0.05）.The numbers of proliferative cells positive for PCNA in experimental group were significantly higher than that in control group at 24 h and 72 h （66.2 ± 4.1 versus 30.0 ±4.8,and 78.7 ± 5.5 versus 54.3 ± 3.9,P 〈 0.05）.Meanwhile,the numbers of apoptotic cells positive for TUNEL staining in experimental group were significantly lower than that in control group at 24 h and 72 h （28.4 ± 3.3 versus 54.5 ± 4.1,and 36.3 ± 3.1 versus 87.4 ± 3.0,P 〈 0.05）.Additionally,peritubular microvascular numbers of experimental animals were 98.3 ± 11.0 and 135.8 ± 13.0,which were significantly higher than 54.4 ± 8.4 and 44.0 ± 6.1 in control ones （P 〈 0.05）.Conclusions Administration of SVF could reduce AKI and accelerate the restoration of damaged kidney after NSS in rats,providing a new strategy for renal protection during NSS.
出处
《中华泌尿外科杂志》
CAS
CSCD
北大核心
2015年第12期925-929,共5页
Chinese Journal of Urology
基金
国家自然科学基金（81370853,81570613）
关键词
脂肪基质血管成分
孤立肾
保留肾单位手术
急性肾损伤
细胞移植
Stromal vascular fraction
Solitary kidney
Nephron-sparing surgery
Acute kidney injury
Cell transplantation
作者简介
通信作者：贾瑞鹏，Email：mipengj@163．com
