冷保存心肌细胞钙循环调控机制及木犀草素作用靶分子研究

心脏移植是解决终末期心脏病的最好方法，但冷保存心脏安全时间短严重限制了它的开展。心肌细胞内钙超载导致心脏心力衰竭是其重要原因，但机制不详。我们的前期研究显示，木犀草素确能改善冷保存大鼠心脏的心肌细胞收缩功能、延长心脏冷保存时间。同时，我们发现：木犀草素能剂量依赖性地降低冷保存大鼠原代心室肌细胞L型钙电流。众所周知，心肌细胞内游离钙离子循环受到细胞膜钙离子通道、内质网钙调节蛋白和线粒体钙转运系统等三个系统调控，但冷保存心肌细胞钙超载的调控过程迄今不详。因此，我们拟制备心肌细胞冷保存模型，应用RT-PCR、膜片钳-激光共聚焦显微技术、Western blot、单细胞检测系统等技术，从细胞-蛋白-RNA 3个层次、细胞膜-内质网-线粒体多角度研究冷保存过程中心肌细胞ca2+循环及其调控蛋白的表达改变规律和木犀草素的作用靶点。本研究的顺利实施将为冷保存心脏心力衰竭的防治提供新思路和新靶点药物。

Cardiac transplantation as the best treatment for end stage cardiac failure has become widely accepted,but the safe heart storage time is still restricted, which have limited its development. As everyone knows, myocardial intracellular calcium overload is the important reason for the heart failure,but the mechanism is unknown. Our previous research has shown that luteolin can indeed improve contractile function of cardiac myocyte with microthermal cardiac solution,extend the preservative time of heart. At the same time, we found: luteolin could dose-related inhibit the L-type calcium currents in microthermal treated ventricular myocytes of the rats.As is known to all,ca2 + circulation in the myocardial cell is controlled by the cell membrane,endoplasmic reticulum calcium regulatory proteins and mitochondrial calcium transport system,but the the mechanism of calcium overload in cardiac myocyte with microthermal cardiac solution is unknown. So we prepare to make a model of cardiac myocyte with microthermal cardiac solution. Then the many techniques such as RT-PCR,patch clamp - laser confocal microscopy,Western blot,cell detection system will be used to research the circulation regulation of ca2 + in cold storaged cardiac myocyte and the target spots witch luteolin act on from the levels of cells-RNA -protein and cell membrane-endoplasmic.This study will provide new ideas of heart preservation and new targets for drugs.