制备家兔离体胸主动脉条改良方法的探讨_临床医学论文.docVIP

制备家兔离体胸主动脉条改良方法的探讨_临床医学论文 制备家兔离体胸主动脉条改良方法的探讨_临床医学论文 作者：杨征　黄燮南 邱敏 吴芹 【摘要】 目的：制备生物活性良好的家兔离体动脉条。方法：以传统的家兔离体动脉条的制备方法为基础，从动物选择、保养液的配置、血管取材、动脉条制备等多个重要环节进行摸索，成功建立了家兔离体动脉条的模型；用家兔主动脉等张收缩的方法，观察去甲肾上腺素（NA）、5-羟色胺（5-HT）和高钾所致离体动脉条收缩反应的影响；应用无Ca2+-复Ca2+的实验方法，以NA作为血管收缩剂，间接观察对细胞内游离钙浓度（[Ca2+]i）的影响。结果：与传统家兔离体动脉条的制备方法相比，改良后NA 10 nmol/L～300 μmol/L、5-HT 10 nmol/L～10 μmol/L、KCl 10～100 mmol/L可致动脉条呈浓度依赖性收缩直至出现最大收缩效应；在无钙Krebs液中，NA 1 μmol/L可致动脉条呈短暂收缩，复钙后等量NA可诱发持续收缩。结论：此方法能使动脉条活性更佳，可作为研究高血压病、抗高血压药物及其相关信号转导通路的有效模型。 【关键词】 动脉条；兔；细胞内游离钙浓度；去甲肾上腺素 Abstract Objective: To prepare thoracic aorta strips of the good bioactivity with rabbits in vitro. Methods: The traditional method was applied by improving several key processes such as selection of animal, preparation of Krebs and aorta strips, and consequently the thoracic aorta strips model of rabbits was successfully established in vitro. The isotonic contraction of the thoracic aorta strips of rabbit was recorded to observe the effects on the concentration-response curves for noradrenaline(NA), 5-hydroxytryptamine(5-HT) and high potassium. The procedure of Ca2+ free- Ca2+ addition was designed to observe indirectly the effects on intracellular free Ca2+ ([ Ca2+]i) by NA. Results: Compared with the traditional method，NA 10 nmol/L-300 μmol/L,5-HT 10 nmol/L-10 μmol/L, KCl 10 -100 mmol/L could induced contraction with a concentraction-dependent manner and the maxium response. NA could induce transient contraction in Ca2+- free medium and the long-lasting by addition of Ca2+ after improving the method. Conclusion: It provides a good model for the study of mechanism and signal transduction pathway of Hypertension and antihypertensive drugs. 　　Key words Aortic strips； Rabbit； Intracellular free calcium； Noradrenaline 高血压病是全世界举世瞩目的流行病，据统计我国现有高血压病患者超过1亿人，并有继续上升的趋势。众所周知，血管对血压的形成和维持具有重要作用，动脉血压过高多半是由于平滑肌过分收缩，引起外周阻力增高所致，因此动脉血管平滑肌是研究高血压病发病机制和防治工作的重要对象[1-2]。离体动脉条的制备是上述研究工作必不可少的环节，但如何制备血管生物活性良好的离体动脉条是研究工作的前提。以传统的家兔离体动脉条的制备为基础，研究动物选择、保养液的配置、血管取材、动脉条制备等因素改良后血管生物活性