梓醇对模拟缺血再灌注损伤兔窦房结细胞凋亡及细胞骨架β―微管蛋白影响.docVIP

梓醇对模拟缺血再灌注损伤兔窦房结细胞凋亡及细胞骨架β―微管蛋白影响 　　摘要：目的 观察地黄主要有效成分梓醇对模拟缺血再灌注损伤兔窦房结细胞凋亡及细胞骨架β-微管蛋白（β-tubulin）的影响，探讨其治疗病态窦房结综合征的机制。方法 取新生乳兔窦房结细胞，分为正常组、模型组及梓醇高、中、低剂量组。以缺氧缺糖模拟缺血、恢复氧和糖的供应模拟再灌注造成窦房结细胞损伤模型。正常组与模型组给予等体积培养基，梓醇高、中、低剂量组给予终浓度为100、20、10 μg/mL药物，应用酶标仪、流式细胞仪、激光共聚焦显微镜观察各组窦房结细胞凋亡率、β-tubulin的形态变化。结果 模型组细胞凋亡率较正常组明显增加（P0.01），β-tubulin裂解明显。与模型组比较，梓醇高、中、低剂量组细胞凋亡率明显降低（P0.01），β-tubulin结构明显完整，荧光强度明显升高（P0.01）。结论 梓醇抑制模拟缺血再灌注引起的窦房结细胞凋亡，保护窦房结β-tubulin形态可能是地黄治疗病态窦房结综合征的机制之一。 　　关键词：梓醇；窦房结细胞；缺血再灌注；细胞凋亡；β-微管蛋白；兔 　　DOI：10.3969/j.issn.1005-5304.2015.08.016 　　中图分类号：R285.5 文献标识码：A 文章编号：1005-5304（2015）08-0059-04 　　Effects of Catalpol on Apoptosis and β-Tubulin of Rabbit Sinoatrial Node Cells in Vitro Injured by Simulated Ischemia Reperfusion LIU Ru-xiu， PENG Jie， LIU Yu， LIU Jin-feng， WANG Yan-li （Guang’anmen Hospital， China Academy of Chinese Medical Sciences， Beijing 100053， China） 　　Abstract：Objective To observe the effects of catalpol， the effective component of Rehmanniae Radix， on atrionector cell apoptosis and β-tubulin in vitro rabbit injured by simulated ischemia reperfusion；To explore the mechanism of treating sick sinus syndrome. Methods Atrionector cells were collected from newborn rabbits. Cells were divided into 5 groups：normal group， model group， catalpol high， medium and low dose groups. Anoxia and aglycaemia were established to simulate ischemia. Atrionector cellular damage models were established by recovering the supply of oxygen and sugar. Normal control group and model group were given the same volume of culture medium， while catalpol high， medium and low dose groups were given medicine with relevant concentrations （100， 20， 10 μg/mL， respectively）. ELISA， FCM， laser scanning confocal microscopy were used to observe apoptosis rate and β-tubulin of atrionector in each group. Results Apoptosis rate of the model group was obviously higher than the normal group （P0.01）， and β-tubulin cleavage was obvious. Apoptosis rate in catalpol high， medium and low dose groups were signifi