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目的:通过酶联免疫吸法检测不同严重程度充血性心力衰竭模型大鼠尿液水通道蛋白2浓度的变化,并与假手术组进行比较。方法:实验于2000-01/2002-01在解放军第一军医大学南方医院动物实验室完成。选用雄性成年SD大鼠42只。取大鼠26只,采用左冠状动脉结扎制备慢性心力衰竭大鼠模型。以左心室梗死面积≥20%为充血性心力衰竭模型造模成功(心力衰竭组,n=13),而左心室梗死面积<20%为心肌梗死心功能代偿组(代偿组,n=13)。其余16只大鼠为对照组:未行冠状动脉结扎。采用BeckmanC×3仪器测定血清钠浓度,采用冰点抑制法测定尿渗量,监测24h尿量。采用双抗体夹心酶联免疫吸附法测定大鼠尿水通道蛋白2浓度。结果:进入结果分析数量保持为42只。①心力衰竭组和代偿组大鼠24h尿量和血钠浓度明显低于对照组(P<0.05～0.01),尿渗量明显高于对照组(P<0.05～0.01)。②对照组大鼠术后4和6周尿液水通道蛋白2水平明显低于其他2组(P<0.05～0.01),代偿组明显低于心力衰竭组(P<0.05,0.01),代偿组和心力衰竭组术后6周明显高于4周(P<0.05)。结论:①水通道蛋白2是心力衰竭时水潴留和低钠血症的关键靶蛋白。②酶联免疫吸附法检测尿液水通道蛋白2浓度,可以有效反映心力衰竭时水潴留和低钠血症状况。
OBJECTIVE: To detect the changes of urinary aquaporin 2 concentrations in rats with congestive heart failure of different severity by enzyme-linked immunosorbent assay (ELISA) and compare with sham operation group. Methods: The experiment was performed at Animal Laboratory of Nanfang Hospital, the First Military Medical University of Chinese PLA from January 2000 to January 2002. 42 male adult SD rats were used. Twenty-six rats were used to establish the chronic heart failure rat model by ligation of the left coronary artery. Left ventricular infarction area ≥ 20% for the congestive heart failure model successful (heart failure group, n = 13), and left ventricular infarction area <20% of myocardial infarction compensatory group (compensatory group, n = 13). The remaining 16 rats as control group: no coronary ligation. Serum sodium concentration was measured by Beckman C × 3 instrument. Urinary osmolality was measured by freezing point inhibition method and urine output was monitored 24h. Urinary aquaporin 2 concentration in rat was measured by double antibody sandwich enzyme-linked immunosorbent assay. Results: The number of entry analysis results remained at 42. ① The 24h urine volume and serum sodium concentration in heart failure group and compensatory group were significantly lower than those in the control group (P <0.05 ~ 0.01), and the urinary osmolality was significantly higher than that in the control group (P <0.05 ~ 0.01). ② The levels of urinary aquaporin 2 in the control group at 4 and 6 weeks after operation were significantly lower than those in the other two groups (P <0.05 ~ 0.01), and the compensatory group was significantly lower than those in the heart failure group (P <0.05 and 0.01) The patients in compensation group and heart failure group were significantly higher than those in 4 weeks after operation (P <0.05). Conclusion: Aquaporin 2 is the key target of water retention and hyponatremia in heart failure. ② ELISA detection of urinary water channel protein 2 concentration, can effectively reflect the state of heart failure, water retention and hyponatremia.