气腹预适应介导内皮祖细胞对大鼠肾脏损伤的作用观察  

Effects of pneumoperitoneum preconditioning on endothelial progenitor cells and renal protective mechanism in rats

作　　者：冯雨明[1] 贾瑞鹏[1] 吴然[1] 吴剑平[1] 周六化[1] 朱佳庚[1] 李文成[1] 

机构地区：[1]南京医科大学附属南京医院(南京市第一医院)泌尿外科,210006

出　　处：《中华医学杂志》2015年第16期1248-1252,共5页National Medical Journal of China

基　　金：国家自然科学基金（81070597）;江苏省卫生厅科教兴卫工程领军人才与创新团队科研课题（IJ201107）;江苏省六大人才高峰资助项目（2011-WS-093）

摘　　要：目的 研究气腹预适应对肾脏损伤的作用及内皮祖细胞（EPC）动员机制.方法 40只SD大鼠采用随机数值表随机分成无气腹组、气腹损伤组（Pp）、长期气腹预适应组（P-L）、中期气腹预适应组（P-M）、短期气腹预适应组（P-S）5组,每组8只.无气腹组气腹压力：0 mmHg（1 mmHg =0.133 kPa）;Pp组气腹压力：15 mmHg,充气60 min;P-L、P-M、P-S组在长时间气腹前给予充气、放气预处理,之后处理同Pp组,P-L组充气时间：25 min,放气时间：10 min;P-M组充气时间：15 min,放气时间：10 min;P-S组充气时间为5 min,放气时间为10 min.术后24 h脱颈椎处死大鼠,取大鼠静脉血及肾组织.血生化及病理学观察大鼠肾功能变化;流式细胞术检测大鼠肾脏EPC归巢特征;免疫组化技术检测大鼠肾脏EPC增殖;Western印迹法检测相关血管源性生成因子的表达.结果 与Pp组比较,P-L、P-M、P-S组大鼠肾功能明显改善[肾小管评分：（1.88±0.35）、（1.63±0.52）、（1.75±0.46）比（2.38±0.52）分,均P＜0.05],P-M、P-S组肾功能改善较P-L明显（均P＜0.05）.流式细胞术检测结果显示,P-L、P-M、P-S组肾脏EPC含量高于Pp组（2.18％±0.14％、2.87％±0.29％、2.90％±0.24％比1.73％±0.19％,均P＜0.05）,其中P-M、P-S组较P-L组更高（均P＜ 0.05）.免疫组化检测结果显示,P-L、P-M、P-S组大鼠肾脏EPC增殖明显高于Pp组,其中P-M与P-S组差异无统计学意义,但较P-L组增强（均P＜0.05）.P-L、P-M、P-S组肾脏组织中可见基质细胞衍生因子1α（SDF-1α）蛋白高表达（均P＜0.05）,其中P-M、P-S组表达更显著（均P＜0.05）.结论 气腹预适应可动员EPC归巢,促进内皮细胞增殖及血管增生,对肾脏损伤产生修复作用.其中,中、短时间预适应方案较长时间预适应可对肾脏产生更佳的保护作用.Objective To explore the pneumoperitoneum-mediated renoprotective effects of preconditioning,mobilizing and homing of endothelial progenitor cells （EPCs） in rats.Methods A total of 40 rats were randomized by a numerical table into 5 groups of gasless （C）,pneumoperitoneum injury （Pp）,long-term pneumoperitoneum preconditioning （P-L）,mid-term pneumoperitoneum preconditioning （P-M） and short-term pneumoperitoneum preconditioning （P-S）.C group had a pneumoperitoneum pressure of 0 mmHg; Pp group 15 mmHg,time 60 min; P-L,P-M,P-S groups were deflated and deflated preconditioning before pneumoperitoneum,then the same as Pp group,P-L group：inflation time was 25 min,gas discharge time 10 min; P-M group：15 min,10 min; P-S group：5 min,10 min.At 24 h postoperation,the animals were sacrificed by destroying cervical spine.And the specimens of venous blood and kidneys were harvested.Also the extent of renal injury,the homing of EPCs,the proliferation and angiogenesis of renal endothelial cell and the expression of angiogenic growth factor were analyzed.Results Compared with Pp group,P-L,P-M and P-S groups exhibited significant improvements in renal function,morphology and histological score（1.88 ±0.35,1.63 ±0.52,1.75 ±0.46 vs 2.38 ±0.52,all P 〈0.05）.The histological scores of P-M and P-S groups improved significantly versus P-L group （both P 〈 0.05）.P-M and P-S groups showed no significant difference in histological score （P 〉 0.05）.The number of EPCs in kidneys increased in P-L,P-M and P-S groups versus Pp group （2.18% ± 0.14%,2.87% ± 0.29%,2.90% ± 0.24% vs 1.73% ± 0.19%,all P 〈 0.05）.The EPCs numbers of P-M and P-S groups were more than that of P-L group （both P 〈 0.05）.And no significant difference existed between P-M and P-S groups （P 〉 0.05）.Compared with Pp group,EPCs of P-L,P-M and P-S groups markedly increased in kidneys.No significant difference existed between P-M and P-S groups,but P-L group was the lowest.Also there was an up-regulated expressio

关 键 词：气腹 人工 适应 生物学 肾 再灌注损伤 内皮祖细胞 

分 类 号：R692[医药卫生—泌尿科学]