七氟醚对大鼠肾缺血-再灌注损伤后水通道蛋白-3的影响

Effect of sevoflurane on aquaroprins 3 after the renal ischemia-reperfusion injury in rat model

目的探讨吸入麻醉剂七氟醚对急性肾缺血-再灌注损伤(IRI)的保护作用及与水通道蛋白-3(AQP3)的相关性。方法SD大鼠随机分为假手术组(C组)、对照组(I/R组)和七氟醚后处理组(Spo组),每组8只。缺血45 min再灌注2 h建立IRI模型。记录术后24 h尿量,检测尿比重。术后24 h,处死大鼠,采血并取肾皮质。测定尿素氮(BUN)和血清肌酐(Cr)水平。免疫组织化学及蛋白质印迹法(Western blot)法对肾组织AQP3进行定性和定量分析。应用SPSS 15.0统计软件分析,数据采用均值±标准差(Mean±SD)表示,采用完全随机设计的单因素方差分析,多组之间的两两比较采用t检验。结果(1)术后24 h尿量,与C组[(5.4±0.2) ml]比较,I/R组[(16.3±9.1) ml]和Spo组[(10.8±5.6) ml]明显增高,差异有统计学意义(I/R组F=11.472,P<0.01,Spo组F=7.429,P<0.05),且Spo组低于I/R组,差异有统计学意义(F=2.120,P<0.05)。术后24 h尿比重,与C组(3.074±0.003)比较,I/R组(3.014±0.012)和Spo组(3.034±0.022)明显降低,差异有统计学意义(I/R组F=188.235,P<0.01,Spo组F=25.964,P<0.01)。(2)与C组[(7.033±0.440) mmol/L]比较,I/R组[(20.530±1.690) mmol/L]和Spo组[(12.300±2.370) mmol/L]血清BUN显著升高,差异有统计学意义(I/R组F=477.867,P<0.01,Spo组F=38.195,P<0.01),且Spo组相对I/R组更低,差异有统计学意义(F=63.952,P<0.05)。与C组比较[(28.7±4.3)μmol/L],I/R组[(122.3±9.5)μmol/L]和Spo组[(82.0±13.2)μmol/L]血清Cr显著升高,差异有统计学意义(I/R组F=644.543,P<0.01,Spo组F=117.922,P<0.01),且Spo组相对I/R组更低,差异有统计学意义(F=49.124,P<0.01)。(3)Western blot及免疫组织化学结果显示,I/R组和Spo组AQP3蛋白均下调,Spo组(419.8±74.2)高于I/R组(138.4±31.4),差异有统计学意义(F=97.586,P<0.05)。结论七氟醚后处理对大鼠急性肾IRI具有保护作用并与上调AQP3明显相关。

Objective To investigate the protective effect of inhaled sevoflurane on renal ischemia-reperfusion injury (IRI) and the its relationship with aquaporins-3 (AQP3) in rat model. MethodsSD rats (n=8) were allocated randomly and equally to operating group (group C), ischemia reperfusion group (group I/R), sevoflurane postconditioning group (group Spo). IRI models were established by 45 min of renal ischemia and 2 h of reperfusion. All animals were killed 24 h after IRI. Serum urea nitrogen (BUN) and creatinine (Cr) were detected. 24 h urine was collected to observe the change of the urine volume and urine specific gravity. The quantity of AQP3 in renal inner medulla was detected by immunohistochemistry and Western blotting. Results(1) 24 h urine volume was significantly increased in group I/R [(16.3±9.1) ml] and group Spo [(10.8±5.6) ml] as compared with group C [(5.4±0.2) ml](Group I/R: F=11.472, P<0.01, Group Spo: F=7.429, P<0.05). The 24 h urine specific gravity was significantly reduced in group I/R (3.014±0.012) and group Spo (3.034±0.022) as compared with group C (3.074±0.003)(Group I/R: F=188.235, P<0.01, Group Spo: F=25.964, P<0.01).(2) BUN in group I/R [(20.530±1.690) mmol/L] and group Spo [(12.300±2.370) mmol/L] was significantly increased as compared with group C [(7.033±0.440) mmol/L](Group I/R: F=477.867, P<0.01, Group Spo: F=38.195, P<0.01). Cr level in group I/R [(122.3±9.5)μmol/L] and group Spo [(82.0±13.2)μmol/L] was significantly increased as compared with group C [(28.7±4.3)μmol/L](Group I/R: F=644.543, P<0.01, Group Spo: F=117.922, P<0.01). The renal function changes in BUN and Cr were significantly improved in group Spo as compared with group I/R (F=63.952, 49.124, P<0.01).(3) As compared with group C (861.6±85.4), the protein expression of AQP3 in group I/R (138.4±31.4) and group Spo (419.8±74.2)(F=505.385, P<0.05) was significantly down-regulated (P<0.05). There protein expression of AQP3 was significant higher in group Spo than group I/R (F=97.586, P<0.01). ConclusionSevoflurane postconditioning could effectively protect renal function against IRI, which was significantly correlated with the up-regulation of of AQP3.