CAJ | 학술논문

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.
목적 연구기복예괄응대신장손상적작용급내피조세포(EPC)동원궤제.방법 40지SD대서채용수궤수치표수궤분성무기복조、기복손상조(Pp)、장기기복예괄응조(P-L)、중기기복예괄응조(P-M)、단기기복예괄응조(P-S)5조,매조8지.무기복조기복압력:0 mmHg(1 mmHg =0.133 kPa);Pp조기복압력:15 mmHg,충기60 min;P-L、P-M、P-S조재장시간기복전급여충기、방기예처리,지후처리동Pp조,P-L조충기시간:25 min,방기시간:10 min;P-M조충기시간:15 min,방기시간:10 min;P-S조충기시간위5 min,방기시간위10 min.술후24 h탈경추처사대서,취대서정맥혈급신조직.혈생화급병이학관찰대서신공능변화;류식세포술검측대서신장EPC귀소특정;면역조화기술검측대서신장EPC증식;Western인적법검측상관혈관원성생성인자적표체.결과 여Pp조비교,P-L、P-M、P-S조대서신공능명현개선[신소관평분:(1.88±0.35)、(1.63±0.52)、(1.75±0.46)비(2.38±0.52)분,균P＜0.05],P-M、P-S조신공능개선교P-L명현(균P＜0.05).류식세포술검측결과현시,P-L、P-M、P-S조신장EPC함량고우Pp조(2.18％±0.14％、2.87％±0.29％、2.90％±0.24％비1.73％±0.19％,균P＜0.05),기중P-M、P-S조교P-L조경고(균P＜ 0.05).면역조화검측결과현시,P-L、P-M、P-S조대서신장EPC증식명현고우Pp조,기중P-M여P-S조차이무통계학의의,단교P-L조증강(균P＜0.05).P-L、P-M、P-S조신장조직중가견기질세포연생인자1α(SDF-1α)단백고표체(균P＜0.05),기중P-M、P-S조표체경현저(균P＜0.05).결론 기복예괄응가동원EPC귀소,촉진내피세포증식급혈관증생,대신장손상산생수복작용.기중,중、단시간예괄응방안교장시간예괄응가대신장산생경가적보호작용.
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 expression of stromal cell derived factor 1-α in pretreated kidneys versus Pp group (all P ＜ 0.05).And P-M and P-S groups increased markedly.Conclusions Pneumoperitoneum-mediated preconditioning protects against kidney injury by promoting EPC homing and enhancing endothelial cell and vascular proliferations.And short and medium-term preconditioning protocols are more effective for protecting kidneys.