目的 研究藏红花酸(crocetin acid,CA)对大鼠实验性房颤(atrial fibrillation, AF)的影响,并探究其离子通道层面的作用机制。方法 用氯化钙-乙酰胆碱给药法诱导体内阵发性AF大鼠模型;使用Langendorff逆行灌流、单酶解消化急性分离大鼠心房肌细胞,应用全细胞膜片钳技术记录、观察和分析不同浓度CA对大鼠心房肌细胞瞬时外向钾通道电流(Ito)的影响。结果 与对照组相比,不同剂量的CA预处理组均延长了模型动物AF诱导时间,降低了AF持续时间。膜片钳实验结果表明,25 μmol·L-1的CA对Ito无显著影响,当其剂量增加到50 μmol·L-1以上时则能显著抑制Ito。其中50、100、200 μmol·L-1的CA可使Ito峰值由给药前(71.10±7.28)pA/pF分别降为(55.49±2.34)pA/pF、(35.56±7.57)pA/pF和(14.69±1.95)pA/pF(P<0.01),表现出明显的浓度依赖性抑制。动力学研究方面,各浓度的CA可使Ito的电流-电压关系曲线(I-V曲线)明显下移,但不改变其激活电位、峰值电位和反转电位。能使稳态激活曲线显著右移,半数激活电压(V1/2-active)降低(P<0.01)。稳态失活曲线显著左移,半数失活电压(V1/2-inactive)降低(P<0.01),而失活后恢复曲线则右移,恢复时间常数(τ)延长(P<0.05)。以上结果显示,CA能够抑制大鼠实验性AF的产生,并抑制Ito的激活、失活及失活后动力学过程。结论 CA具有抗药物诱导的AF作用,此效应可能与其抑制Ito,并改变其动力学过程有关。
Abstract
OBJECTIVE To study the effect of crocetin acid(CA)on experimental atrial fibrillation in rats and the mechanism of effect at the ion channel level. METHODS Rat model of paroxysmal atrial fibrillation(AF)was induced by administration of calcium chloride-acetylcholine. Atrial myocardial was digested by single enzymatic perfusate on the Langendorff device. Whole-cell patch clamp techniques was used to record the effects of different concentrations of CA on Ito and its kinetics in rat atrial myocytes. RESULTS Compared with the control group, all groups of CA pretreatment prolonged the induction of AF and reduced the duration of AF. The results of the patch clamp experiment showed that CA had no significant effect on Ito at 25 μmol·L-1. When the concentration of CA was increased to above 50 μmol·L-1, the peak of Ito was significantly inhibited, decreasing from(71.10±7.28) pA/pF to(55.49±2.34)pA/pF,(35.56±7.57)pA/pF, and(14.69±1.95) pA/pF by CA of 50 μmol·L-1 , 100 μmol·L-1, and 200 μmol·L-1, respectively (P<0.01), in a concentration-dependent manner. CA down-regulated the current-voltage relationship(I-V)curve of Ito, but activation potential, peak potential, and reversal potential of Ito had no change. CA shifted the activation curve of Ito to the depolarized direction, with significantly decreased half activation voltage(V1/2-a)(P<0.01). CA shifted the inactivation curve of Ito to the hyperpolarized direction, with significantly decreased half inactivation voltage(V1/2-in)(P<0.01). In addition, CA significantly prolonged the recovery constant time(τ)after Ito inactivation(P<0.05). These results indicated that CA could alleviate experimental atrial fibrillation in rats and inhibit the current density, activation, inactivation, and recovery processes of Ito. CONCLUSION CA has an antagonism effect on drug-induced atrial fibrillation by inhibiting Ito and regulating gating kinetic of Ito.
关键词
藏红花酸 /
瞬时外向钾通道 /
心律失常 /
心房颤动 /
全细胞膜片钳 /
心房肌细胞
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Key words
crocetin acid /
transient outward potassium current /
arrhythmia /
atrial fibrillation /
patch clamp /
atrial myocyte
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中图分类号：
R966
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参考文献
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脚注
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基金
国家自然科学基金项目资助(82174129);扬州大学大学生科技创新基金重点项目资助(X20210726)
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