葛根素对缺血/再灌注损伤兔肺组织天冬氨酸特异性半胱氨酸蛋白酶-3变化的影响  

Change of caspase-3 in rabbits after lung ischemia-reperfusion and the effect of puerarin(葛根素)

作　　者：许益笑[1] 王万铁[1] 徐正祄[1] 金晓凤[1] 郝卯林[1] 

机构地区：[1]温州医学院病理生理学教研室,浙江温州325035

出　　处：《中国中西医结合急救杂志》2008年第2期104-107,F0003,共5页Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care

基　　金：浙江省温州市科技计划项目(Y2005A080)

摘　　要：目的探讨天冬氨酸特异性半胱氨酸蛋白酶-3(caspase-3)在兔肺缺血/再灌注损伤(LIRI)中的变化及葛根素的保护作用机制。方法健康日本大耳白兔30只,随机分为对照组、缺血/再灌注(I/R)组及葛根素组,每组10只。复制单侧LIRI模型。对比观察各组血清超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量、一氧化氮(NO)含量、肺湿/干重(W/D)比值及肺泡损伤指数(IQA);用原位末端缺刻标记法(TUNEL)测定肺组织细胞凋亡指数(AI);用免疫组化及原位杂交技术分别检测肺组织caspase-3蛋白和mRNA表达。结果与对照组比较,I/R组血清SOD活性和NO含量降低,MDA含量、W/D比值、IQA、AI值及caspase-3表达均上调(P均<0.01)。与I/R组比较,葛根素组SOD、NO显著升高,MDA、W/D比值、IQA、AI值及caspase-3表达明显下降(P均<0.01)。相关分析显示:AI值与SOD、NO呈明显负相关,与MDA、caspase-3 mRNA及caspase-3蛋白呈明显正相关(P均<0.01);caspase-3 mRNA与SOD、NO呈明显负相关,与MDA含量呈明显正相关(P均<0.01)。结论葛根素可通过提高体内NO水平、降低氧自由基水平、减轻脂质过氧化反应,下调caspase-3的mRNA和蛋白表达,从而抑制肺再灌注后肺组织细胞的异常凋亡,减轻LIRI。Objectiye To investigate the change of caspase-3 in rabbits after lung ischemia-reperfusion injury （LIRI） and the effect of puerarin （葛根素）. Methods Thirty healthy rabbits used for unilateral lung ischemia-reperfusion model were randomly divided into 3 groups （each n= 10）： control group （C group）, lung ischemia-reperfusion group （I/R group） and puerarin group. The activity of serum superoxide dismutase （SOD）, the contents of serum malondialdehyde （MDA） and nitric oxide （NO）, the wet to dry weight （W/D） ratio of lung tissue and the index of quantitative assessment of histological lung injury （IQA） were measured respectively in different groups; the pneumocyte apoptosis index （AI） was achieved by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling （TUNEL）; caspase-3 protein and mRNA expression were studied by using in situ hybridization （ISH） and immunocytochemistry （IHC） techniques in the groups mentioned above. Results The activity of SOD and content of NO were significantly lower in I/R group than those in C group （both P〈0.01）; while the content of MDA, the value of W/D, IQA and AI, together with the expression of caspase-3 mRNA and protein were evidently higher （P〈0.01, respectively）. In the comparisons between puerarin group and I/R group, it was shown that the content of MDA, the value of W/D, IQA, AI and expression of caspase-3 were decreased （P〈0. 01, respectively）, while the activity SOD and the content of NO were increased in the former group than those in the latter group （P〈0. 01, respectively）. Result of correlation analysis： AI was significantly negatively correlated with the activity SOD and the content of NO, and significantly positively correlated with the content of MDA, caspase-3 mRNA and caspase-3 protein （all P〈0.01）. Caspase-3 mRNA was significantly negatively correlated with SOD activity and NO content, and markedly positively correlated with MDA content （all P〈0.01）. Conclu

关 键 词：缺血/再灌注损伤 肺 凋亡 天冬氨酸特异性半胱氨酸蛋白酶-3 葛根素 

分 类 号：R285.5[医药卫生—中药学] R363[医药卫生—中医学]