生长反应因子1特异的脱氧核酶与大鼠颈动脉损伤后诱生型一氧化氮合酶表达的关系  

Association of DNA enzyme targeting Egr-1 with the expression of inducible nitric oxide synthase in rats after balloon injury

作　　者：滕雅轩[1] 刘闺男[1] 周敬[1] 

机构地区：[1]中国医科大学附属第一医院循环内科,辽宁省沈阳市110001

出　　处：《中国临床康复》2005年第34期52-55,i0004,共5页Chinese Journal of Clinical Rehabilitation

基　　金：辽宁省教育厅高等学校科学技术研究项目(2004D185)~~

摘　　要：目的:观察生长反应因子1特异的脱氧核酶ED5对大鼠颈动脉血管损伤后诱生型一氧化氮合酶表达的影响,探讨生长反应因子1特异的脱氧核酶对血管损伤后内膜增生的抑制作用。方法:实验于2004-03/2004-07在中国医科大学实验动物部完成。选用健康雄性Wistar大白鼠96只。随机将动物分为4组:治疗组、稀释液对照组、转染液对照组和假手术组,每组24只,每组分为4个时间点(3,7,14,21d)进行观察。①血管球囊损伤的动物模型制作:麻醉后,切开鼠颈正中,钝性分离左颈动脉,血管夹暂时阻断左颈总动脉近心、远心端血流,用2F导管从颈外动脉插入左颈总动脉内,推入0.2mL生理盐水充盈气囊,反复抽拉球囊3次,造成左颈总动脉内膜损伤。②分组干预:假手术组只进行颈动脉结扎,但不插入导管,即不造成动脉内膜损伤。稀释液对照组、转染液对照组、治疗组均在造成血管球囊损伤模型的同时,局部注射200μL的1mmol/LMgCl2液;局部注射含30μL的FuGENE6Reagent的MgCl2液200μL;颈外动脉给予含有异硫氰酸荧光素标记的生长反应因子1特异的脱氧核酶500μg+转染试剂FuGENE630μL的MgCl2液200μL。③诱生型一氧化氮合酶mRNA水平检测:采用半定量反转录聚合酶链反应。④诱生型一氧化氮合酶蛋白合成检测:采用免疫印迹法。⑤血管内膜增生情况观察:苏木精-伊红染色,光镜显微镜下观察,应用计算机图像分析软件测定血管内膜和中膜厚度。⑥血管内膜和中膜一氧化氮合酶蛋白表达检测:采用免疫组织化学法。⑦统计学分析:多组间比较采用单因素方差分析,两组间比较采用LSD-t检验。结果:大鼠96只均进入结果分析。①大鼠血管内皮损伤后血管病理形态学改变:内皮损伤后3d内膜增生不明显,7d内膜开始增生,14和21d时内膜增生明显。②血管组织诱生型一氧化氮合酶mRNA的表达及蛋白合成结果:假手术组大鼠颈动脉无�AMI： To observe the effects of DNA enzyme targeting Egr-I（EDS） on the expression of inducible nitric oxide synthase （iNOS） in rats with injury of carotid artery and study the mechanism of ED5 in inhibiting neointimal hyperplasia. METHODS： The experiment was done in the Experimental Animal Center of China Medical University during March and July 2004. Totally 96 Wistar rats were randomly divided into four groups： treatment group, dilution solution group, transfection solution group and sham operation group with 24 rats in each group. Observation in each group was conducted at days 3, 7, 14 and 21. ①Establishment of animal models of balloon injury： under anesthesia, a incision was cut in the center of rat＇s neck to separate left carotid artery through blunt dissection. Vascular clamp was used to block blood flow of proximal and distal end from left carotid artery to heart. Endothelial denudation of rats was performed with 2F balloon catheter. ② Grouping and intervention： Sham operation group was only undertaken carotid artery ligation without inserting tube. Rats in the dilution solution group, transfection solution group and treatment group were locally injected with 200 μL of 1 rmnol/L MgC12, 200μL MgC12 containing 30 μL FuGENE6 Reagen, and 500 μg ED5＋200μL MgC12 containing 30μL FuGENE6, respectively. ③Measurement of iNOS mRNA： Semi-quantitative reverse transcription-polymerase chain reaction（RT-PCR） was adopted. ④Measurement of iNOS synthesis： Western blotting was adopted. ⑤ Observation of neointimal hyperplasia： Haematine-eosin staining was used for observation under light microscope and thickness of intima and media were calculated by using computer. ⑥Expression of iNOS protein in intima and media of vessel： Western blotting was adopted. ⑦Statistical analysis： One-way analysis of variance and LSD-t test were used. RESULTS： All the 96 rats were involved in the result analysis. ①Pathomorphological changes after vascular endothelial in iury： Neointimal

关 键 词：颈动脉/病理学 增生 一氧化氮合酶 血管内膜 

分 类 号：R311.8[医药卫生—基础医学]