THE RESEARCH FOR THE ATHEROSCLEROSIS AND THE CHANGE OF CELLULAR SIGNALING WITH REFERENCE TO THE AGING OF VASCULAR ENDOTHELIAL CELL

本研究通过测定Na^+-K^+ ATP酶活性、乙基异丙氨氯吡嗪敏感性Na^+/H^+逆向转运蛋白活性、细胞内游离钙离子浓度（[Ca^++]i），使用荧光指示剂fura-2，使用荧光指示剂2 7 -bis测定细胞内pH（[pH]i）（羧乙基）羧基荧光素（BCECF）和在人脐静脉内皮细胞（HUVEC）中掺入^3H-胸苷合成DNA。因此，我们研究了内皮素、蛋白激酶C（PKC）激活剂佛波酯、PKC抑制剂H-7、Na^+离子载体莫能菌素、细胞外Ca^+<++>和电压依赖性钙通道（VDC）阻断剂对这些参数的影响。并且部分依赖于细胞外Ca^<++>或电压依赖性Ca^<++>通道。内皮素可促进HUVEC产生前列环素和EDRF，但对[pH]i和[Ca^++]i无影响，这种作用与平滑肌细胞收缩有拮抗作用。此外，内皮素还具有抑制细胞增殖的作用，这一现象提示存在自身调节机制，提示内皮素不仅对血管收缩有促进作用，而且对动脉硬化也有促进作用。

In this research we can measure the activity of Na^+-K^+ ATPase, the activity of ethylisopropylamiloride sensitive Na^+/H^+ antiporter, the cytosolic free calcium ion concentration ([Ca^<++>]i) using a fluorescent indicator fura-2, the intracellular pH ([pH]i) using a fluorescent indicator 2 7 -bis(carboxyethyl) carboxyfluoresein (BCECF) and the synthesis of DNA using the incorporation of ^3H-thymidine in human umbilical vein endothelial cells(HUVEC). Therefore we investigated the effect of endothelin, a protein kinase C(PKC) activator phorbol ester, a PKC inhibitor H-7, Na^+ ionophore monensin, extracellular Ca^<++> and a voltage dependent calcium channel(VDC) blocker on these parameters.The [pH]i in HUVEC is 7.17, and the maintenance of the [pH]i depends mainly on the ethylisopropylamiloride sensitive Na^+/H^+ antiporter, and depends partly on the extracellular Ca^<++> or the voltage dependent Ca^<++> channel. Endothelin enhanced the prostacyclin generation and the EDRF generation in HUVEC without effects on [pH]i and [Ca^<++>]i, these effects are antagonistic to the contraction for the smooth muscle cells. Furthermore endothelin has the suppression of the cell proliferation, this phenomenon suggests the presence of the autoregulatory mechanism, through these results it is suggested that endothelin has the acceleration effect on not only the vascular contraction but also the arteriosclerosis.