摘要
目的评价聚腺苷二磷酸核糖聚合酶-1(PARP-1)在大鼠肺缺血再灌注损伤中的作用及其与自噬的关系.方法清洁级健康雄性成年SD大鼠24只,体重200~240 g,8~12周龄,采用随机数字表法分为3组(n=8):假手术组(S组)、缺血再灌注组(I/R组)和缺血再灌注+PARP-1抑制剂组(I/R+PJ34组).S组仅开胸不阻断左肺门;I/R组和I/R+PJ34组采用阻断左侧肺门45 min,再灌注120 min的方法制备大鼠肺缺血再灌注损伤模型;I/R+PJ34组于缺血前30 min腹腔注射PJ3410 mg/kg,S组和I/R组给予等容量生理盐水.于再灌注120 min时取左肺组织,称重并计算湿重/干重(W/D)比值;采用HE染色法观察肺组织病理学结果;采用TUNEL法观察细胞凋亡情况,计算细胞凋亡指数;采用Western blot法检测缺血区肺组织PARP-1活化产物PAR、Bcl-2、Bax、微管相关蛋白1轻链3(LC3)和Beclin-1的表达,计算Bcl-2/Bax比值和LC3-Ⅱ/LC3-Ⅰ比值.结果与S组比较,I/R组和I/R+PJ34组肺组织W/D比值、病理学损伤评分、细胞凋亡指数和LC3-Ⅱ/LC3-Ⅰ比值升高,PAR和Beclin-1表达上调,Bcl-2/Bax比值降低(P<0.05);与I/R组比较,I/R+PJ34组肺组织W/D比值、肺组织病理学损伤评分、细胞凋亡指数和LC3-Ⅱ/LC3-Ⅰ比值降低,PAR和Beclin-1表达下调,Bcl-2/Bax比值升高(P<0.05).结论 PARP-1的激活参与了大鼠肺缺血再灌注损伤的过程,其机制可与增加自噬诱发细胞凋亡有关.
Objective To evaluate the role of poly( ADP-ribose) polymerase-1 ( PARP-1) in lung ischemia-reperfusion ( I/R ) injury in rats and the relationship with autophagy. Methods Twenty-four clean-grade healthy male Sprague-Dawley rats, aged 8-12 weeks, weighing 200-240 g, were divided into 3 groups (n=8 each) using a random number table method: sham operation group (S group), lung I/R group ( I/R group) and lung I/R plus PARP-1 inhibitor PJ34 group ( I/R+PJ34 group). The chest was only opened without clamping the left hilum of lung in group S. Lung I/R injury model was established by clam-ping the left hilum of lung for 45 min followed by 120 min reperfusion in I/R and I/R+PJ34 groups. PJ3410 mg/kg was intraperitoneally injected at 30 min before ischemia in group I/R+PJ34, while the equal volume of normal saline was injected in S and I/R groups. The rats were sacrificed at the end of reperfusion, and lungs were removed for microscopic examination of the pathological changes ( with a light microscope) which were scored and for determination of wet to dry weight ratio ( W/D ratio), cell apoptosis ( by TUNEL), ex-pression of PARP-1 activity markers ( PAR), Bcl-2, Bax, microtubule-associated protein 1 light chain 3Ⅰ( LC3-Ⅰ), LC3-Ⅱ and Beclin-1 ( using Western blot ). The apoptosis index, Bcl-2/Bax ratio and LC3-Ⅱ/LC3-Ⅰ ratio were calculated. Results Compared with S group, the W/D ratio, pathological scores, apoptosis index and LC3-Ⅱ/LC3-Ⅰ ratio were significantly increased, the expression of PAR and Beclin-1 was up-regulated, and Bcl-2/Bax ratio was decreased in I/R and I/R+PJ34 groups (P<0. 05). Compared with I/R group, the W/D ratio, pathological scores, apoptosis index, and LC3-Ⅱ/LC3-Ⅰratio were significantly decreased, the expression of PAR and Beclin-1 was down-regulated, and Bcl-2/Bax ratio was increased in I/R+PJ34 group ( P<0. 05). Conclusion PARP-1 activation is involved in lung I/R inju-ry in rats, and the mechanism may be related to increasing autophagy and inducing cell apoptosis.
作者
邢娜
邢飞
渠明翠
李艳娜
李平乐
张建文
张卫
Xing Na;Xing Fei;Qu Mingcui;Li Yanna;Li Pingle;Zhang Jianwen;Zhang Wei(Department of Anesthesiology,First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052,China)
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2019年第6期746-749,共4页
Chinese Journal of Anesthesiology
基金
河南省高等学校重点科研项目(19A320010).
