肢体缺血再灌注致兔肺组织损伤及葛根素的干预作用

Lung injury of rabbit induced by reperfusion of limb ischemia and the interference effect of puerarin

目的观察肢体缺血再灌注致兔肺组织损伤时抗氧化系统的变化,细胞凋亡数、Bc l-2和Bax蛋白表达,及葛根素的干预作用。方法健康家兔60只,随机分3组:A组(n=20)为假手术组,B组(n=20)为缺血再灌注组;C组(n=20)为葛根素治疗组。实验结束时,取肺组织测定超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)、黄嘌呤氧化酶(XOD)活性和丙二醛(MDA)、一氧化氮(NO)含量,同时计算肺系数及进行肺病理学检查,应用原位缺口末端标记技术(TUNEL)和免疫组织化学方法,观察肢体缺血4h再灌注4h肺脏细胞凋亡数及Bc l-2和Bax蛋白的表达。结果肢体缺血4h再灌注后4h肺组织SOD、GSH-PX活性和NO含量显著下降,XOD活性和MDA含量明显增高,肺系数升高,肺组织病变明显。葛根素的干预治疗可明显缓解上述变化。假手术对照组肺脏见极少量凋亡细胞,缺血再灌注组、葛根素治疗组肺脏均可见大量凋亡细胞,但葛根素治疗组凋亡细胞数比缺血再灌注组明显减少。假手术对照组肺脏组织Bc l-2呈阳性表达,基本未见Bax蛋白表达,其余2组可见大量肺脏组织出现Bc l-2和Bax蛋白阳性表达,但葛根素治疗组肺脏的Bax蛋白阳性表达明显较缺血再灌注组减少。结论家兔肢体缺血再灌注后,肺抗氧化功能下降、肺脏细胞凋亡增多及Bc l-2、Bax蛋白表达上调,葛根素的干预治疗对这种肺损伤有明显的保护作用。

Objective To investigate the change of antioxidation system, the number of cell apoptosis and the expression of Bax and Bcl-2 in lung injury of rabbit after reperfusion of limb ischemia and the interference effect of puerarin. Methods Sixty rabbits were randomly divided into 3 groups. Group A were sham-operated animals ; group B were ischemia-reperfusion animals; group C, animals treated by puerarin. At the end of the experiment, the animals were killed and the left lung extraced in order to detected the antioxidant status of lung tissue. Lung index was calculated as a maker of edema. The lung histopathologic changes in experimental rabbits were observed. Immunohistochemieal studies and TUNEL were done to evaluate cell apoptosis. The activity of SOD and GSH-PX as well as XOD , the content of MDA and NO as well as the expressions of Bax and Bcl-2 in lung tissue were determined. Results In the rabbit model with bilateral hind limb （IR） injury, the SOD, GSH-PX activity and NO content were significantly decreased, and the XOD activity , MDA level and lung index were increased. Conversely, rabbits treated with puerarin before the onset of the ischemic period, the XOD and MDA levels were significantly reduced,as well as SOD , GSH-PX and NO levels elevated , and pulmonary edema significantly less . In addition, the histopathologic detection showed that more severe lesion in IR group rabbits than the rabbits treated with puerarin. Puerarin could suppress XOD activity, reduce the MDA production, inhibit Bax expression and cell apoptosis, and induce Bcl-2 expression in lung tissue. Conclusion Puerarin has protective effect on lung injury of rabbit with limb ischemia-reperfusion which is due to its elevating the ability of antioxidisity and its anti-lipid peroxidation reaction.