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白藜芦醇对豚鼠心室肌细胞L
作者:易方方,唐其柱,冯斯婷,王腾【关键词】 白藜芦醇;全细胞膜片钳;钙电流
Effects of resveratrol on Ltype calcium current in isolated ventricular myocytes of guinea pig
【Abstract】 AIM: To investigate the effects of resveratrol on the Ltype calcium current (ICaL) in isolated ventricular myocytes of guinea pig. METHODS: Single ventricular cell was obtained from healthy guinea pig heart using enzymatic dissociation methods. The wholecell patch clamp was used to record the ICaL following the administration of resveratrol at 1, 50, 100 μmol/L. RESULTS: ① Resveratrol reduced the ICaL in a concentrationdependent manner, but it had no effect on IV shape of ICaL. ② Resveratrol markedly shifted the steadystate activation curve of ICaL to the right, and V1/2 value increased from (-18.97±4.08) mV in control to (-15.97±4.13) mV in the presence of resveratrol (n=7, P=0.02), the κ value from (3.64±0.56) to (3.82±0.60) (n=7,P=0.67). ③ Inactivation curve of ICaL was not affected significantly. ④ The time constant of reactivation of ICaL was reduced from (98.50±9.00) ms to (54.53±4.27) ms (n=6,P=0.000) by resveratrol. CONCLUSION: Resveratrol inhibited the Ltype calcium channel, mainly acting on the activity and recovery processes.
【Keywords】 resveratrol;wholecell patch clamp;calcium current
【摘要】 目的:研究白藜芦醇对正常豚鼠心室肌细胞L型钙电流(ICaL)的影响. 方法:采用全细胞膜片钳技术,记录不同浓度白藜芦醇作用前后对ICaL的影响. 结果:①白藜芦醇呈浓度依赖性阻断ICaL,但对激活电位、峰电位、反转电位均没有影响;②白藜芦醇使ICaL稳态激活曲线右移,V1/2由对照的(-18.97±4.08) mV变为(-15.97±4.13) mV (n=7,P=0.02),κ由(3.64±0.56)变为(3.82±0.60)(n=7,P=0.67);③对失活曲线无影响,不改变ICaL失活特性;④白藜芦醇使ICaL失活后再恢复时间常数τ缩短,从(98.50±9.00) ms变为(54.53±4.27) ms(n=6,P=0.000). 结论:白藜芦醇对ICaL有阻断作用,主要作用于钙通道激活和恢复过程.
【关键词】 白藜芦醇;全细胞膜片钳;钙电流
0引言
白藜芦醇化学名为3,5,4′三羟基反二苯代乙烯(3,5,4′ trihydroxyrans stilbene),是一种主要存在于葡萄皮、花生中的多酚类植物抗毒素. 现认为它是适量饮用葡萄酒有利于心血管健康的主要作用因子[1]. 大量研究[2-3]发现白藜芦醇对心肌缺血-再灌注损伤有保护作用;对血管有广泛的舒张效应[4];能抗动脉粥样硬化,防治冠心病[5]. 但其对心肌细胞离子通道的作用少见报道,我们采用膜片钳技术研究白藜芦醇对心室肌细胞ICaL的影响.
1材料和方法
1.1材料健康成年豚鼠31只(武汉大学医学院动物中心),体质量250~350 g,雌雄不拘; I型胶原酶,蛋白酶E,HEPES(Gibco公司); CSOH, CSCl, Na2ATP(Sigma公司);白藜芦醇(西安赛得高科生物股份有限公司);其余均为国产分析纯. 无钙台氏液 (mmol/L):NaCl 135,KCl 5.4,MgCl2 1,NaH2PO4 0.33,Glucose 20,HEPES 10,pH 7.38 (NaOH调节);KB液(mmol/L): glumatic acid 50,KOH 85,KCl 30,MgSO4 3,Taurine 20,Glucose 10,HEPES 10,EGTA 0.5,pH 7.2(KOH调节);电极外液组成(mmol/L):NaCl 37,Choline Chloride 100,CaCl2 1.8,MgCl2 0.5, HEPES 10,Glucose 10,CSCl 4.6,pH 7.3(CSOH调节);电极内液组成(mmol/L): CSCl 120, CaCl2 1.0, MgCl2 5, Na2ATP 5,EGTA 11, HEPES 10,Glucose 11,pH 7.4(CSOH调
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