雌激素对心肌L型钙通道及钠钙交换体的调控作用及其机制分析-药理学专业论文.docxVIP

华中科技大学同济医学院博士学位论文 华中科技大学同济医学院 博士学位论文 - - 6 - The regulation and the mechanism of estrogen on L type calcium and sodium calcium exchanger in cardiomyocytes Department of Pharmacology, Tongji Medical College, HUST Candidate: Yang Xiao-yan Advi so r: Prof. Zeng Fan-dian Abstract Objective: Long QT Syndromes (LQTs) is a cardiac disorder characterized by a prolongation of the QT interval on ECG. LQTs predispose the hearts to develop the potentially lethal ventricular tachyarrhythmia, called torsades de pointes (TdP), which cause episodic syncope and sudden death. Women are known to be at higher risk to congenital and acquired form of TdP, but in adolescents（human 14 years old and rabbit42 days old）before the surge of sex steroids, the risk of TdP is reserved with male being more susceptible to TdP. Some evidences suggest that estrogen may be a contributory factor for the greater prevalence of drug-induced TdP in women. Sex-differences in the propensity to early afterdepolarizations (EADs) and TdP are reversed in adolescence and correlate with higher levels of L type Ca2+ channels (Ca-L) and current, ICa-L, and sodium calcium exchangers (NCX) also play a role in sex difference of arrhythmia phenotype. In this study, we analyzed the sex, age and regional difference in NCX and studied the regulation and mechanism of estrogen on Ca-L and NCX in rabbit cardiomyocytes. Method: (1) Single left ventricular myocardial cells of rabbits were isolated by enzymic method. According to the aim of experiment, myocytes were incubated with DMSO, 17-β-Estradiol (E2) or other reagents. ICa-L was recorded by the whole-cell - - PAGE 7 - configuration of voltage clamp thechnique. The level of Cav1.2 (α1C) protein was measured by immunocytochemistry and the level of Cav1.2 (α1C) mRNA was measured by single-cell quantitative real time RT-PCR. To evaluate the effect of estrogen on dofetilide-induced EAD, action potential (AP) was recorded under current-clamp mode, and then dofetilide was added t