丹参对大鼠肺缺血再灌注损伤保护作用的研究

1、丹参对大鼠肺缺血再灌注损伤保护作用的研究         11-02-13 10:43:00     编辑：studa20                         作者：程键，程青，杨剑虹，陆卫华 【摘要】  目

2、的： 探讨丹参对大鼠肺缺血再灌注损伤的保护作用并探讨其机制。方法：60只Wistar大鼠随机分为3组：手术对照组(A)，缺血再灌注组(B)，丹参干预组(C)。每组分别于缺血第45 min、再灌注30 、60、120 min，观察肺组织病理形态变化，检测血浆丙二醛(MDA)、超氧化物歧化酶(SOD)的含量，测定肺组织湿/干重(W/D)比值，TUNEL法测定肺组织中细胞凋亡指数(AI)。结果:(1)肺组织病理变化：B组肺组织毛细血管充血、水肿、炎性细胞浸润显著，C组肺组织较B组减轻;(2)血浆MDA、SOD含量：经缺血再灌注后，B和C组血浆MDA较A组均明显增加(P<0.05)，B组增加的程

3、度明显高于C组(P<0.05);B和C组血浆SOD较A组同时点均显著下降(P<0.05)，C组减少的程度明显小于B组(P<0.05);(3)肺组织W/D 比值：经缺血再灌注后，B组和C组的肺组织W/D比值都呈进行性上升(再灌注60，120 min vs缺血45 min，P<0.05);C组上升幅度明显小于B组(再灌注60，120 min时，C组vsB组，P<0.05);(4)肺组织细胞AI的变化：B组及C组缺血再灌注后均可见肺组织细胞凋亡现象，但与B组相比，C组相同时点的肺组织细胞凋亡数显著减少(P<0.05)。结论:丹参对于实验性大鼠肺缺血再灌注损伤具有一

4、定的保护作用，其作用机制与清除氧自由基、抗氧化及抗细胞凋亡有关。 【关键词】  丹参;再灌注损伤;肺缺血;大鼠            A study of the protective effects of radix salviae miltiorrhizae on ischemiareperfusion injury in rat lungCHENG Jian, CHENG Qing, YANG Jianhong, LU Weihua(Department of Emer

5、gency, General Hospital of Guangzhou Military Area in Wuhan, Wuhan 430070, China)ABSTRACT Objective: To study the mechanism of the protective effects of radix salvia miltiorrhiza on lung ischemiareperfusion injury.Methods: Sixty Wistar rats were randomly divided into three groups: surgical control g

6、roup (group A)，ischemiareperfusion group (group B) and Radix salvia miltiorrhiza group (group C).In each group, 5 rats were sacrificed at one of four time points(45 min after ischemia;30 min，60 min and 120 min after reperfusion).The concentration of malondialdehyde(MDA)and superoxide dismutase(SOD)i

7、n plasma were detected，and the lung tissue wet/dry (W/D) ratio were examined.The histology of lung was observed，and the apoptotic index (AI) was detected by deoxynucleotidyl transferasemediated dUTP nick end labeling (TUNEL).Results: (1).Histological changes：Marked pulmonary capillary congestion，ede

8、ma and infiltration were observed in group B, while less pathologic alterations were shown in group C. (2).The concentration of MDA in plasma of both group B and C were significant higher at each time point after ischemiareperfusion than that of group A (P<0.05). Between group B and C, the increa

9、sed level of the former group was obviously higher than the latter one (P<0.05); The concentration of SOD in plasma of group B and C were significantly lower at each time point than that of group A(P<0.05)，and the decreased level of group C was obviously lower than that of group B (P<0.05).

10、 (3)The W/D ratio of lung tissue in group B and C increased progressively with time after reperfusion (60 min, 120 min reperfusion vs 45 min ischemia，P<0.05);but the increase degree of group C was obviously lower than that of group B (at 60 min, 120 min after reperfusion， group C vs group B，P<

11、0.05).(4). The AI of lung tissue were observed in both group B and group C, but AI in group C at different time points after reperfusion were all significant less than that in group B (P<0.05).Conclusion: radix salvia miltiorrhiza can effectively protect Wistar rats from ischemiareperfusion lung

12、injury and the mechanism is involved with its function of oxyradical elimination，antioxygen and antiapoptosis.KEY WORDS Radix salvia miltiorrhiza; Reperfusion injury; Lung ischemia;Wistar rats近年来,随着心脏手术、器官移植、心肺脑复苏措施的深入开展，体外循环技术在临床上的应用日益增多，对肺缺血/再灌注(lung ischemiareperfusioniniury， LIRI)损伤，及其所导致的肺功能障碍的

13、发生和防治也逐渐被人们重视。LIRI的确切发病机制尚未完全明了，目前认为与氧化损伤、钙超载、中性粒细胞引起的过度炎症反应及细胞凋亡等有关。众多实验研究表明,中药能较好地抗LIRI，具有良好的应用前景，其中，丹参尤为受到广泛关注。本实验旨在通过建立大鼠在体LIRI模型，在缺血再灌注之前给予丹参，研究丹参对LIRI是否具有保护作用及其作用机制。1 材料与方法1.1 动物与试剂健康雄性Wistar大鼠60只，体重(200±50) g，由华中科技大学同济医学院动物中心中提供。丹参注射液由四川升和制药有限公司生产，国药准字Z51021303。超氧化物歧化酶(Superoxide dismuta

14、se，SOD)、丙二醛(malondialdehyde，MDA)检测试剂盒均购置于南京建成生物工程公司;细胞凋亡检测试剂盒(POD)购置于武汉博士德生物工程公司。1.2 动物模型制备健康Wistar大鼠，3%戊巴比妥钠50 mg/kg腹腔注射麻醉后，尾静脉注射肝素(100 U/kg)。气管切开后进行气管插管，小动物呼吸机辅助通气。胸骨右缘第四肋间开胸，结扎右主支气管和右肺动静脉。并应用温热的37 的生理盐水棉纱敷盖创口。缺血的时间为45 min，为缺血期。45 min后放开结扎线，使肺脏充盈后，计时120 min，为再灌注期。再灌注期结束后左房放血处死大鼠。1.3 动物分组及处理60只大鼠随机分成3组，每组20只。(1)手术对照组(A组)：预先给予生理盐水0.5 mL/kg腹腔内注射，开胸游离右肺门后，不行夹闭阻断;(2)缺血再灌注组(B组)：预先给予生理盐水0.5 mL/kg腹腔内注射，开胸游离右肺门后，夹闭阻断45 min，继行再灌注;(3)丹参干预组(C组)：预先丹参5 g/kg腹腔内注射，余操作同B组。各组分别于夹闭缺血45 min、再灌注后30，60，120