微透析技术实时检测肾热缺血后葡萄糖变化的动物实验研究

目的探讨应用微透析技术研究兔肾热缺血再灌注损伤后肾皮质葡萄糖浓度变化规律的可行性。方法新西兰大耳白兔20只,其中实验组10只,麻醉后游离出右肾及其动静脉,将微透析探针置入肾背侧皮质内,复方氯化钠溶液持续灌注平衡60min后,连接电化学分析系统,阻断肾动静脉60min,然后开放动静脉,观察60min。利用电化学方法实时在线检测肾透析液中葡萄糖含量的变化。对照组10只,仅游离出肾动静脉而不结扎,同样留置微透析针取样,连续检测葡萄糖浓度,比较2组在正常灌注期、缺血期和再灌注期葡萄糖浓度的变化。结果葡萄糖电极的电流响应与其浓度呈良好的线性关系,微透析针的回收率在（63.6±2.1）%。实验组兔肾皮质正常状态下测得的透析液葡萄糖浓度为（1.89±0.37）mmol/L,缺血期葡萄糖浓度为（0.69±0.12）mmol/L（LSD检验,P=0.000）,缺血期较自身正常灌注期降低（36.7±2.4）%;再灌注期葡萄糖浓度为（0.62±0.14）mmol/L（LSD检验,P=0.000）。与对照组比较,实验组缺血期（t=-11.975,P=0.000）和再灌注期（t=-11.993,P=0.000）葡萄糖浓度显著降低。结论应用微透析与活体在线电化学技术相结合的方法可以方便、灵敏检测肾缺血再灌注损伤后葡萄糖水平,可以较好地实时反映肾皮质缺血状态。

Glucose Metabolism in the Warm Ischemia/Reperfusion Injured Kidney of Rabbits: In Vivo Microdialysis and Real-Time Electrochemical Detection

Objective To investigate the change of glucose metabolism in warm ischemia/reperfusion injured kidney of rabbits in vivo by microdialysis and real-time electrochemical detection. Methods A total of 20 New Zealand rabbits were divided into experiment and control groups （ 10 in each）. After anesthesia by intravenous injection of harbitone, an oblique incision under the 12th rid was made in the animals to expose the right kidney and the right renal artery and vein. Afterwards, a microdialysis probe was placed into the dorsal renal cortex along the long axle of the kidney. After 60 minutes to equilibrate, an electrochemical detection system was used to continuously determine the concentration of glucose in the renal cortex before ischemia. Then, ischemia/reperfusion injury model was established by clamping the renal pedicle for 60 minutes followed by perfusion for 60 minutes. The control group received the same intervention without clamping the renal pedicle. The glucose concentration of the microdialysis samples were analyzed before, during, and after the isehemia. Results The reaction of the glucose electrode to the electric current was linearly correlated with the concentrations of glucose, and the rate of microdialysis probe recycling was （63.6 ± 2. 1 ） %. The concentration of glucose in the renal cortical interstitial samples were （ 1.89 ±0.37） , （0. 69 ±0. 12） , and （0. 62 ±0.14） mmol/L respectively before, during, and after the warm ischemia. During the ischemia phase, the mean concentration of glucose decreased by （36. 7 ±6 2.4） % from the base level （ LSD test, P = 0. 000）. The glucose concentration of the experimental group was significantly lower than that in the control at both ischemia and reperfusion phases （ t = - 11. 975, P = 0. 000 ; t = - 11. 993, P = 0. 000, respectively）. Conclusions In vivo microdialysis combined with electrochemical detection provides a sensitive and real-time method for measuring the glucose concentration in warm ischemia/reperfusion injured kidney, which can suggest the ischemic condition of the renal cortex.