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目的:研究17-甲氧基-7-羟基-苯并呋喃查尔酮(YLSC)对H2O2诱导的心肌细胞内钙超载的拮抗作用及对L型钙电流(ICa-L)的影响。方法:采用SD大鼠乳鼠进行心肌细胞培养,实验分为①正常对照组;②H2O2组:上机前加入终浓度为0.3 mmol.L-1的H2O2;③预先给予低、中、高不同终浓度YLSC药物处理组:分别给予100,200,400μmol.L-1YLSC的无血清培养基,孵育24 h,上机前加入终浓度为0.3 mmol.L-1H2O2,以Fluo-3/AM荧光指示剂负载,应用激光共聚焦显微镜技术,分别于加入H2O2后15 min内检测细胞内[Ca2+]i的变化;分离昆明种小鼠单个心室肌细胞,全细胞膜片钳技术记录YLSC对ICa-L的影响。结果:①与正常对照组比较,H2O2诱导的模型组细胞内[Ca2+]i增加60.43%±7.75%,而高、中、低YLSC预处理组[Ca2+]i分别增加38.39%±13.87%,14.49%±2.94%,-28.1%±1.52%,与模型组比较显著降低(P<0.01)。②YLSC可使心室肌细胞ICa-L的电流-电压(I-V)关系曲线上移,能改变ICa-L的激活和失活特征,使ICa-L的激活曲线和稳态失活曲线左移。结论:YLSC对心肌细胞钙离子通道有较好的阻断作用,能显著减轻H2O2诱导的心肌细胞内[Ca2+]i超载。
AIM: To investigate the antagonism of 17-methoxy-7-hydroxy-benzofuran chalcone (YLSC) on H2O2-induced intracellular calcium overload and its effect on L-type calcium current (ICa-L). Methods: SD rat neonatal rat cardiomyocytes were cultured and divided into ① normal control group; ②H2O2 group: H2O2 at a final concentration of 0.3 mmol.L-1 was added before the machine was added; ③At the end of the experiment, Concentration YLSC drug-treated groups: 100, 200, 400μmol.L-1YLSC serum-free medium was given and incubated for 24 h. The cells were loaded with 0.3 mmol.L-1H2O2 at the final concentration of 0.3 mmol.L-1H2O2 and loaded with Fluo-3 / AM fluorescent indicator. Laser confocal microscopy was used to detect the intracellular [Ca2 +] i within 15 min after addition of H2O2. Single ventricular myocytes of Kunming mice were isolated and the whole cell patch clamp technique was used to record the effect of YLSC on ICa-L. Results: Compared with the normal control group, [Ca2 +] i increased by 60.43% ± 7.75% in H2O2-induced model group compared with the control group, while the [Ca2 +] i increased by 38.39% ± 13.87%, 14.49 % ± 2.94%, -28.1% ± 1.52%, which were significantly lower than those in model group (P <0.01). (2) YLSC up-regulated the current-voltage (I-V) curve of ICa-L in ventricular myocytes, and changed the activation and inactivation of ICa-L, leaving the activation curve and steady-state inactivation curve of ICa-L to the left. CONCLUSION: YLSC can block the Ca (superscript 2 +) channel in cardiomyocytes and reduce the [Ca2 +] i overload induced by H2O2.