白藜三醇对快速刺激离体兔心房氧化应激的影响  

The effect of resveratrol against atrial oxidative stress injury induced by rapid-pacing in isolated rabbit hearts

作　　者：胡晓芹[1] 王志荣[1] 张超群[1] 张卓琦[1] 孙红[2] 程明月[1] 

机构地区：[1]徐州医学院附属医院心内科,江苏徐州221006 [2]徐州医学院生理教研室,江苏徐州221002

出　　处：《中国心脏起搏与心电生理杂志》2014年第1期50-54,共5页Chinese Journal of Cardiac Pacing and Electrophysiology

基　　金：江苏省兴卫工程医学重点人才资助项目(项目编号:RC20070809)

摘　　要：目的观察白藜三醇(RES)对快速刺激离体兔右房自由基代谢的影响,探讨其心脏保护作用的可能机制。方法 32只家兔建立离体心脏Langendorff灌流模型,电极放置右心耳处,利用BL-420S生物机能系统以电压4.0 V,频率600次/分进行刺激,随机分为对照组(不起搏),快速起搏组(RAP组),还原型烟酰胺腺嘌呤二核苷酸氧化酶(NADPH)抑制剂夹竹桃麻素组(APO组),RES组,每组8只。对照组离体灌流70 min,不起搏;RAP组离体灌流30 min,快速起搏心房40 min;APO、RES组分别离体药物处理30 min,快速起搏心房40 min。灌流结束后,留存心房肌组织。比色法测定超氧化物岐化酶(SOD)、丙二醛(MDA)、体内活性氧(ROS)、NADPH活性,HE染色检测病理变化,免疫组化半定量分析calpain1的表达,用western blot方法检测calpain1、cTnT和gp91phox蛋白的表达,逆转录聚合酶链式反应方法检测calapin1、p22phox和gp91phox的mRNA表达。结果①与对照组比较,RAP、APO、RES组MDA、ROS和NADPH的表达均增加,SOD含量减少;与RAP组比较,APO、RES组MDA、ROS和NADPH表达均降低,SOD含量增加(P均<0.05);而APO组与RES组比较,相关氧化应激指标无差异;②对照组心肌细胞排列整齐;RAP组心肌细胞排列紊乱、稀疏,组织结构破坏;APO组心肌细胞排列较紧凑;RES组心肌细胞较APO组排列稍紊乱、稀疏;③与对照组比较,RAP、APO、RES组calpain1、gp91phox蛋白及mRNA表达和p22phox的mRNA表达均增加,cTnT蛋白表达降低(P<0.05);与RAP组比较,RES、APO组calpain1、gp91phox蛋白及mRNA表达和p22phox的mRNA表达均减低,cTnT蛋白表达增加(P<0.05);但与APO组比较,RES组相关蛋白及mRNA表达无差异。结论 RES可能通过部分抑制NADPH氧化酶活性,从而减少氧自由基的生成,进而减轻快速起搏右房导致的氧化应激损伤。Objective To investigate the protective effects of resveratrol （RES） on oxidative stress injury induced by rapid-pacing in isolated rabbits. Methods The experiment was performed on 32 isolated rabbit hearts perfused on Lan- gendorff apparatus, and taken use of BL-420S biological function of system with stimulation of voltage 4.0V, frequency 10 Hz. They were randomized into 4 groups： Control group, Rapid atrial pacing group （RAP）, Apocynin pretreatment group （APO） , Resveratrol group（RES） , with 8 in each group. The Control group was perfused with Krebs-Henseleit buffer（ K- H） solution at 4℃ for 70 min, The RAP group was firstly perfused with K-H solution for 30 min and then rapid pacing on the right atrium of the isolated rabbit hearts for 40 min, The APO group and the RES group were pretreated with apocynin and resveratrol for 30 min and then rapid pacing on the right atrium of the isolated rabbit hearts for 40 min. At the end ofall the groups perfused, the atrial tissue were obtained and stored for the further study. The activity of superoxide dis- mutase（SOD）, Maleie Dialdehyde（MDA）, ROS and re- duced nicotinamide adenine dinucleotide phosphate oxi- dase in tissue from RA were measured; HE staining was used to observe the pathological changes; The immuno- histochemical and western blot were used to detective theexpression of the proteins ; The reverse transcription-PCR was used to detective the expression of the mRNA. Result ① Compared with the Control group, MDA, ROS and NADPH oxidase expression in the RAP, RES and the APO group in- creased （P〈0.01）, the activity of SOD in the three groups were reduced （P〈0.01） ; With the RAP group, MDA, ROS and NADPH oxidase expression in the APO and the RES group were lower （ P〈0.05 ）, the content of SOD increased （ P〈 0.05 ） ; While APO group compared with the RES group, the relevant indicators of oxidative stress were no significant differences（P〉0.05 ） ;②Atrial myocytes with the normal structure of the nor

关 键 词：心血管病学 氧化应激 还原型烟酰胺腺嘌呤二核苷酸氧化酶 快速刺激 离体兔 白藜三醇 

分 类 号：R331.38[医药卫生—人体生理学]