血管内皮生长因子受体1在七氟烷预处理减轻大鼠心肌缺血再灌注损伤中的作用:离体实验

Role of vascular endothelial growth factor receptor 1 in reduction of myocardial ischemia reperfusion injury by sevoflurane preconditioning in rats

目的通过离体大鼠心肌缺血再灌注损伤模型,探讨血管内皮生长因子受体1(vascular endothelial growth factor receptor 1, VEGFR1)在七氟烷预处理减轻心肌缺血再灌注损伤中的作用。方法采用随机数字表法将48个SPF级雄性SD大鼠心脏分为4组(n=12):对照组(Sham组)、心肌缺血再灌注组(I/R组)、七氟烷预处理组(SevoPC组)和VEGFR1拮抗剂组(MF1组)。采用Langendorff灌流装置60mmHg恒压灌注30分钟,停止灌注45分钟,恢复灌注120分钟建立离体大鼠心肌缺血再灌注损伤模型。观察恢复灌注120分钟期间,心脏左室发展压(left ventricular developed pressure, LVDP)、左室舒张末压(left ventricular end-diastolic pressure, LVEDP)、左室内压最大上升速率(+dp/dtmax)和左室内压最大下降速率(-dp/dtmax)的变化情况;检测恢复灌注120分钟后心肌梗死面积、冠脉流出液中cTnI(cardiac troponin-I, cTnI)、TNF-α(tumor necrosis factor-α, TNF-α)和IL-6 (interleukin 6, IL-6)含量,以及心肌VEGFR1表达。结果与I/R组比较,SevoPC组再灌注期间LVDP和±dp/dtmax上升、LVEDP下降;再灌注120分钟后心肌梗死面积减少[(17.6±2.3)%v.s.(31.3±4.2)%;t=3.698,P=0.011]、冠脉流出液中cTnI含量减少[(52.4±5.1)v.s.(110.7±4.5)ng/ml;t=4.125, P<0.001]、TNF-α水平降低[(16.8±1.7)pg/ml v.s.(37.6±2.3)pg/ml;t=3.917,P <0.001]和IL-6水平降低[(11.6±1.5)v.s.(18.5±1.4)pg/ml;t=3.602,P=0.012],心肌VEGFR1表达上调[(1.05±0.06)v.s.(0.32±0.05);t=4.165, P <0.001]。与Sevo PC组比较,MF1组再灌注期间LVDP和±dp/dtmax下降、LVEDP上升;再灌注120分钟后心肌梗死面积增加[(33.6±5.1)%v.s.(17.6±2.3)%;t=3.974,P=0.002],冠脉流出液中c Tn I含量增加[(124.3±3.9) v.s.(52.4±5.1)ng/ml;t=3.846,P <0.001]、 TNF-α水平升高[(33.7±1.6)v.s.(16.8±1.7)pg/ml;t=3.707,P <0.001]和IL-6水平升高[(19.4±1.1) v.s.(11.2±1.5)pg/ml;t=3.589,P=0.016],心肌VEGFR1表达降低[(0.21±0.03)v.s.(1.05±0.06);t=4.215, P<0.001]。结论七氟烷预处理可能通过上调心肌VEGFR1的表达,抑制心肌炎性反应减轻大鼠心肌缺血再灌注损伤。

Objective To investigate the role of vascular endothelial growth factor receptor 1 （VEGFR1） in reduction of myocardial ischemia reperfusion（I/R） injury by sevoflurane preconditioning. Methods Forty-eight isolated adult male rat hearts were randomly divided into 4 groups （n=12 each） sham operation group （Group Sham）,ischemia reperfusion（I/R） injury group（Group I/R）, sevoflurane preconditioning group （Group SevoPC）. VEGFR1 inhibitor group （Group MF1） using Langendorff heart assay. In Group Sham, isolated hearts were treated with preoxygenated KH constant perfusion for 195 min. In Group I/R, isolated hearts suffered ischemia for 45 min after 30 min-perfusion and received reperfusion for 120 min so as to establish I/R models. In Group SevoPC, isolated hearts were treated by KH with 3% sevoflurane constant perfusion for 15 min after 15 min-KH perfusion. In group MF1, isolated hearts were treated by KH with 10 μmol/L MF1 and KH with 3% sevoflurane constant perfusion for 15min. Cardiac function parameters were measured during the whole period of reperfusion including left ventricular end-diastolic pressure （LVEDP）, left ventricular developed pressure （LVDP）, maximum LVDP increase rate （dp/dtmax）, and maximum LVDP decrease rate （-dp/dtmax） . The release of cardiac troponin I （cTnI）, tumor necrosis factor-α （TNF-α） and interlrukin 6 （IL-6） in coronary effluent, myocardial infarct size, the cardiac infarction size, and the expression of VEGFR1 in myocardium were evaluated after 120-min reperfusion. Results Compared with Group I/R, the cardiac functions in Group SevoPC significantly improved （LVDP ↑ , LVEDP ↓,±dp/dtmax ↑ ） during 120 min-reperfusion. In Group SevoPC, the infarct size [（17.6±2.3） % v.s. （31.3±4.2） %], the level of cTnI [（52.4±5.1） v.s. （110.7±4.5） ng/ml], TNF-α [（16.8±1.7） v.s. （37.6±2.3） pg/ml] and IL-6 [（11.6±1.5）v.s. （18.5±1.4） pg/ml] reduced by up-regulation of VEGFR1[（1.05±0.06）v.s. （0.32±0.05）].Compared with Group SevoPC, cardiac functions （LVDP ↓ , LVEDP ↑ , ±dp/dtmax ↓ ） in Group MF1 were impaired during 120 min-reperfusion. Moreover, in Group I/R, myocardial infarct size[（33.6±5.1） % v.s. （17.6±2.3） %], the level of cTnI [（124.3±3.9） v.s. （52.4±5.1） ng/ml], TNF-α[（33.7±1.6）v.s. （16.8±1.7） pg/ml]and IL-6[（19.4±1.1） v.s. （11.2±1.5） pg/ml]increased by down-regulation of VEGFR1[（0.21±0.03） v.s. （1.05±0.06）]. Conclusion Sevoflurane preconditioning alleviates cardiac ischemia reperfusion injury in rats via up-regulating of VEGFR1 and anti-inflammation.