大鼠肾脏外科性缺血时间与急性肾损伤的剂量-反应曲线特征研究  

Dose-response curves characteristics between surgical ischemia time and renal injury

作　　者：王雷[1] 陈健文[2] 李怀康 李修彬 徐俊楠 焦启龙 袁清 黄燕 黄庆波[1] 王保军[1] 马鑫[1] Wang Lei;Chen Jianwen;Li Huaikang;Li Xiubin;Xu Junnan;Jiao Qilong;Yuan Qing;Huang Yan;Huang Qingbo;Wang Baojun;Ma Xin(Department of Urology,the Third Medical Centre,Chinese PLA(People′s Liberation Army)General Hospital,Beijing 100039,China;Department of Nephrology,the First Medical Center,Chinese PLA General Hospital,Chinese PLA Institute of Nephrology,Beijing 100853,China)

机构地区：[1]解放军总医院第三医学中心泌尿外科医学部,北京100039 [2]解放军总医院第一医学中心肾脏病医学部解放军肾脏病研究所,北京100853

出　　处：《中华实验外科杂志》2021年第11期2204-2207,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81970665、82100713、81970594、81802804);解放军总医院"国家杰出青年"培育专项(2020-JQPY-002);解放军总医院军事医学青年专项(QNF19068)。

摘　　要：目的探索大鼠肾脏的热缺血时间与急性肾损伤的剂量-反应曲线特征。方法建立大鼠单侧肾脏缺血再灌注急性损伤模型(切除对侧肾脏),设置20、24、28、32、36、40、44 min缺血时间组及假手术组,通过血清生化检测、PAS组织切片过碘酸雪夫染色、原位缺口末端标记法(TUNEL)凋亡检测、活性氧水平检测、透射电镜观察等方法评估急性肾损伤程度,绘制缺血时间与急性肾损伤的剂量-反应曲线,组间资料的差异比较采用独立样本t检验。结果 36 min组血清肌酐[(344.60±58.59) μmol/L比(94.10±28.91) μmol/L,t=9.393,P<0.01]、尿素氮[(49.33±15.24) mmol/L比(17.74±6.92) mmol/L,t=4.622,P<0.01]、急性肾小管坏死(ATN)评分[(3.86±0.65)分比(1.38±0.35)分,t=8.158,P<0.01]、TUNEL凋亡评分(0.59±0.06)分比(0.22±0.09)分,t=8.577,P<0.01]、活性氧阳性细胞比例[(0.74±0.13)比(0.23±0.06),t=7.977,P<0.01]、丙二醛含量[(0.75±0.66) nmol/mg比(0.51±0.11) nmol/mg,t=3.828,P<0.01]均显著高于32 min组;36 min组线粒体损伤高于32 min组。32 min及以下的缺血时间组无损伤或为轻度损伤,40、44 min缺血时间组均为重度损伤。结论大鼠肾脏外科性缺血诱发急性肾损伤的剂量-反应曲线为S形,曲线在32~36 min之间陡峭上升,32 min以内的大鼠单侧肾脏血流阻断不诱发重度急性肾损伤,造成重度急性肾损伤存在缺血时间"临界点"。Objective To investigate the characteristics of dose-response curve of warm ischemia time that causes acute kidney injury in mice.Methods Unilateral kidney ischemia-reperfusion injury mouse model(with contralateral nephrectomy)was established,different ischemia time(20,24,28,32,36,40,44 minutes)were selected for each group.After 24 h reperfusion,acute kidney injury was evaluated via functional and histological analysis,and reactive oxygen species(ROS)production level.comparison was performed using student′s t-test.Results Functional analysis indicated that,both serum creatinine and blood urea nitrogen(BUN)level in 36 minutes group was significantly increased when compared with 32 minutes group[serum creatinine,(344.60±58.59)μmol/L vs.(94.10±28.91)μmol/L,t=9.393,P<0.01;BUN,(49.33±15.24)mmol/L vs.(17.74±6.92)mmol/L,t=4.622,P<0.01].Histological analysis showed that,36 minutes group exhibited more heavily pathological tubular injury:tubular injury score in 36 minutes and 32 minutes group were 3.86±0.65 and 1.38±0.35,respectively(t=8.158,P<0.01);and terminal-deoxynucleotidyl transferase mediated nick end labeling(TUNEL)positive cells in 36 minutes and 32 minutes group were 0.59±0.06 and 0.22±0.09(t=8.577,P<0.01).ROS positive cell proportion in 36 minutes group was significantly higher than that in 32 minutes group(0.74±0.13 vs.0.23±0.06,t=7.977,P<0.01).And the concentration of malondialdehyde(MDA)in 36 minutes group was significantly higher than that in 32 minutes group[(0.75±0.66)nmol/mg vs.(0.51±0.11)nmol/mg,t=3.828,P<0.01].All the results above in 20,24,28 minutes exhibited much more slightly damage when compared with 40,44 minutes ischemia group.Conclusion Dose-response curve of WIT on acute renal injury was S-shaped with a steep ascending phase between 32 and 36 minutes,reperfusion that not excess of 32minutes did not observed severe acute kidney injury,and there is a threshold ischemia time in severe renal injury.

关 键 词：急性肾损伤 缺血时间"临界点" 氧化应激 剂量-反应曲线 

分 类 号：R699.2[医药卫生—泌尿科学]