离体大鼠冠状动脉的分离培养及张力测定技术  

Measurement techniques of isolation,organ culture and isometric tension of rat coronary artery in vitro

作　　者：邝素娟[1] 邓春玉[1] 杨慧[1] 饶芳[1] 刘晓颖[1] 张梦珍[1] 朱杰宁[1] 单志新[1] 林秋雄[1] 杨敏[1] 余细勇[1] 

机构地区：[1]广东省人民医院(广东省医学科学院)医学研究中心、广东省心血管病研究所,广东广州510080

出　　处：《中国药理学通报》2014年第3期137-141,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81273516,81070103,81330007);广东省自然科学基金重点项目(NoS2012020010947);广东省自然科学基金资助项目(NoS2013010014762)

摘　　要：目的建立体外培养大鼠冠状动脉和测定血管平滑肌舒缩功能的方法,为研究各种药物对血管张力的作用提供良好的血管模型.方法采用脱臼法处死SD大鼠（200～250g）,取下心脏,冰浴条件下显微操作分离冠状动脉,制备成1.8～2.0mm长的血管条,置于含DMEM-F12培养基（内含10%胎牛血清、1×105U·L-1青霉素、100mg·L-1链霉素）的培养皿内,在37℃,通以95%O2、5%CO2混合气的条件下培养24h后,用两根不锈钢丝穿过血管腔,固定于微血管张力测定仪的浴槽内.分别给予血管收缩剂5HT、U46619、CaCl2和血管舒张剂ACh,然后测定冠状动脉的张力变化.结果培养的冠状动脉对5-HT、U46619、CaCl2产生浓度依赖性收缩,对ACh产生浓度依赖性舒张反应.结论采用离体器官培养法培养离体的冠状动脉环,能保持血管平滑肌基本的功能,可用于各种因素对血管结构和功能影响的研究.Aim To establish organ culture technique of rat coronary artery and provide an applicable model for the study of effects of various drugs on vascular tone. Methods Male Sprague-Dawley rats(200 ~ 250 g-weight) were killed by cervical vertebra dislocation,and the coronary arteries were rapidly dissected out with micromanipulation under the sterile condition of ice bath. After removal of fat and adventitia,the coronary arteries were cut into rings(1. 8 ~ 2. 0mm in length) for organ culture,and the vessels were cultured in Dulbecco's Modified Eagle Medium-F12 containing 10% fetal bovine serum,1 × 105U ·L- 1penicillin and 100 mg · L- 1streptomycin for 24h at 37℃with humidified 95% O2and 5% CO2. Rings of coronary arteries were mounted in a small vessel myography using two stainless steel wires for the measurement of isometric tension. Effects of the vasoconstrictors(5-HT,U46619 and CaCl2) and the vasodilator(ACh) on coronary arterial rings were determined. Results5-HT,U46619 and CaCl2could produce a concentration-dependent contraction response,and ACh could induce a concentration-dependent relaxation response in rat coronary artery after organ culture. Conclusion Rat coronary artery cultured in vitro can maintain normal function of the vascular smooth muscle and can be used to investigate the effects of vasoactive agents.

关 键 词：冠状动脉 培养 血管张力 大鼠 血管收缩剂 血管舒张剂 

分 类 号：R332[医药卫生—人体生理学] R322.121[医药卫生—基础医学]