壳聚糖纳米粒载体转染铜超氧化物歧化酶对缺血再灌注脑组织的作用  

Effects of chitosan nanoparticles loaded the copper-superoxide dismutase plasmid on brain tissue after ischemia-reperfusion

作　　者：贾栋[1] 高国栋[1] 李永林[1] 

机构地区：[1]解放军第四军医大学唐都医院神经外科,陕西省西安市710038

出　　处：《中国组织工程研究与临床康复》2008年第45期8827-8830,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:由于血脑屏障的存在,铜超氧化物歧化酶在临床应用中往往达不到其在体外对中枢神经细胞缺氧、复氧过程中的自由基清除效果。那么能不能利用壳聚糖纳米粒这一种新的基因载体来增强铜超氧化物歧化酶的治疗效果呢?目的:观察壳聚糖纳米粒-超氧化物歧化酶联合体对脑缺血再灌注脑组织氧自由基的清除能力。设计:观察实验。材料:铜超氧化物歧化酶质粒由解放军第四军医大学唐都医院中心实验室提供。36只大鼠制备缺血再灌注模型,随机分为3组:生理盐水对照组;生理盐水+超氧化物歧化酶组;壳聚糖超氧化物歧化酶质粒治疗组。方法:以分子交联法制备壳聚糖纳米粒;通过静电吸附作用连接上超氧化物歧化酶质粒;将50.0μL纳米粒分别与10.0,25.0,50.0,75.0,100.0μL超氧化物歧化酶质粒混合,经琼脂糖凝胶电泳分析壳聚糖纳米粒与超氧化物歧化酶DNA的结合能力,并通过紫外分光光度计检测其包埋率。将生理盐水、超氧化物岐化酶、壳聚糖纳米粒-超氧化物歧化酶质粒联合体(体积比50∶75)输注到缺血再灌注大鼠体内,然后测定大鼠脑海马组织内丙二醛水平及超氧化物歧化酶活性。主要观察指标:壳聚糖纳米粒与超氧化物歧化酶质粒的结合情况。各组大鼠脑组织中丙二醛水平和超氧化物歧化酶活性。结果:壳聚糖纳米粒:超氧化物歧化酶质粒体积比为50∶75时包埋率最大,为100%,且此种比例下有良好的DNA保护作用,不被DNaseI降解。经壳聚糖纳米粒-超氧化物歧化酶质粒联合体治疗的大鼠,其海马组织丙二醛水平明显低于生理盐水对照组及生理盐水+超氧化物歧化酶组(P<0.01和P<0.05),超氧化物歧化酶活性明显高于生理盐水对照组及生理盐水+超氧化物歧化酶组(P<0.01和P<0.05)。结论:壳聚糖纳米粒-超氧化物歧化酶质粒联合体在脑组织中具有较强氧自由基清除能力。BACKGROUND： The copper-superoxide dismutase （Cu-SOD） can not play an effective role in free radical-scavenging in the process of hypoxia or reaeration because of blood-brain barrier, whether the treating effect can be enhanced by chitosan nanoparticles is uncertain. OBJECTIVE： To observe the ability of chitosan nanoparticles loaded the Cu-SOD plasmid on free radical-scavenging in ischemia-reperfusion brain tissue. DESIGN： An observation experiment. MATERIALS： The Cu-SOD plasmid was provided by the Center Laboratory of Tangdu Hospital, Forth Military Medical University of Chinese PLA. A total of 36 rats were prepared ischemia-reperfusion models, and randomly divided into the control, physiological saline （PS） plus Cu-SOD plasmid and chitosan nanoparticles loaded the Cu-SOD plasmid groups. METHODS： The chitosan nanoparticles were prepared by cross-linking method. The Cu-SOD plasmid was absorbed to chitosan nanoparticles by electrostaficforces, mixed the 50.0 μL nanoparticles with 10.0, 25.0, 50.0, 75.0, 100.0 μL SOD plasmid respectively. The electrophoretic analysis was used to examine the combination condition of the chitosan nanoparticles with superoxide dismutase plasmid, and the embedding rate was calculated with ultraviolet spectrophotometer. Chitosan nanoparticles, SOD and Cu-SOD plasmid were infused into the rat after brain iscbemia-reperfusion. Further more, to detect the content of malondialdehyde and superoxide dismutase in brain tissues. MAIN OUTCOME MEASURES： The combinb.tion condition of the chitosan nanoparticles with superoxide dismutase plasmid, and the activity of malondialdehyde （MDA） and SOD were measured. RESULTS： Chitosan nanoparticles and superoxide dismutase in volume ratio of 50：75 could harvest the largest embedding rate with 100%, which had good protective effect on DNA. The contents of MDA in brain tissues of the Chitosan nanoparticles loaded the Cu-SOD plasmid group were obvious lower than that of the control （P 〈 0.01） and the physiological saline plu

关 键 词：壳聚糖 纳米粒 Cu—SOD质粒 脑缺血再灌注 氧自由基 

分 类 号：R318[医药卫生—生物医学工程]