参附注射液对大鼠肺缺血再灌注损伤肺组织的保护作用

Protective effect of shenfu injection on lung injury induced by pulmonary ischemia reperfusion in rats

目的:研究参附注射液(shenfu injection,SF)对大鼠肺缺血再灌注损伤(lung ischemia reperfusion injury,LIRI)中肺组织细胞凋亡的保护作用,并探讨其可能机制。方法:雄性SD大鼠42只,随机分为假手术组(Sh组)、肺缺血再灌注组(IR组)和参附注射液组(SF组),建立在体单侧肺缺血再灌注模型,缺血45min后再灌注3、6h。SF组于阻断肺门前30min腹腔注射SF10mL/kg。于实验结束点取肺组织,分别以Annexin-V-PI双染法通过流式细胞仪检测细胞凋亡率,免疫组化二步法检测caspase-3表达,同时测定丙二醛(MDA)浓度和超氧化物歧化酶(SOD)活性,以及光镜和电镜检查。结果:与Sh组比,IR组肺组织细胞凋亡率、caspase-3表达、MDA浓度升高,SOD活性下降,差异均有显著性(P<0.01);SF组较IR组细胞凋亡率、caspase-3表达、MDA浓度下降,SOD活性升高,差异亦均有显著性(P<0.01)。形态学观察发现SF组肺组织损伤明显轻于IR组。结论:SF可能通过下调caspase-3表达而阻止细胞凋亡,从而减轻LIRI中肺组织损伤。

Objective To investigate the protective effect of shenfu injection （SF） on lung injury induced by lung ischemia reperfusion in rats. Methods Unilateral lung in vivo ischemia reperfusion model in rats was established by clamping the left pulmonary hilum for 45 minutes and then releasing of 3 and 6 hours, respectively. Forty-two male Sprague-Dawley rats were divided randomly into three groups： sham operation group（Sh group）, ischemia reperfusion group （IR group） and shenfu injection group （SF group）. Each group was redivided into two subgroups of 3 hours and 6 hours. Shenfu injection （10 mL/kg） was intraperitoneally injected 30 minutes before ischemia in SF group. The apoptosis rate in lung tissue was detected by the way of Annexin-V-PI in flow cytometer. The expression of caspase-3 was measured by immunohistochemical stain, and the mean optical density（OD） values of positive fields were examined quantitively by image analysis system. The concentration of malondialdehyde （MDA） and the activity of superoxide dismutase （SOD） were obtained. The morphological and ultrastructure changes of lung tissue were also observed. Results Compared with IR group, the apoptosis rate, the expression of caspase-3 and the concentration of MDA of long tissue were significantly decreased in SF group but higher than that in Sh group （P 〈 0.01）, and the activity of SOD was obviously increased in SF group but lower than that in Sh group （P 〈 0.01）. The pathological and ultrastructure changes were better in SF group than in IR group, too. Conclusion SF may effectively inhibit the apoptosis rate of lung tissue during lung ischemia reperfusion injury, and extenuate the lung injury degree by the possible mechanism of decreasing the concentration of MDA, increasing the activity of SOD and reducing the expression of caspase-3 in early period of lung ischemia reperfusion injury.