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目的:观察通阳活血方对模拟缺血再灌注损伤兔窦房结细胞凋亡及骨架蛋白Desmin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,以缺氧缺糖模拟缺血,以恢复氧和糖的供应模拟再灌注造成窦房结细胞损伤模型。正常对照组与模型组给予等体积培养基,通阳活血方高、中、低剂量组分别给予相应浓度药物(终浓度分别为100、20、10μg/m L),运用流式细胞仪、激光共聚焦显微镜观察各组窦房结细胞凋亡率、细胞骨架蛋白Desmin形态的变化。结果:模型组细胞凋亡率较正常组明显升高(P<0.01);细胞骨架蛋白Desmin裂解明显。通阳活血方高、中、低剂量组细胞凋亡率明显低于模型组(P<0.01),Desmin平均荧光强度明显高于模型组(P<0.01)。结论:通阳活血方治疗病态窦房结综合征的机制可能与对抗缺血再灌注所致的窦房结细胞凋亡及保护细胞骨架蛋白Desmin形态结构有关。
Objective: To observe the effect of Tongyang Huoxue Decoction on the apoptosis of sino-atrial node cells and the Desmin protein of skeletal muscle in simulated ischemia-reperfusion injury rabbits and to explore its mechanism of treating sick sinus syndrome. Methods: The sinoatrial node cells of neonatal rabbit rabbits were mimicked with hypoxia and hypoglycemia to simulate the reperfusion of the sinoatrial node cells by restoring the supply of oxygen and sugar. The normal control group and model group were given equal volume of medium, Tongyang Huoxue Fang high, middle and low dose groups were given the corresponding concentration of drugs (final concentrations were 100,20,10μg / m L), using flow cytometry, laser Confocal microscopy was used to observe the apoptosis rate of sinoatrial node cells and the change of Desmin morphology in each group. Results: The apoptotic rate of model group was significantly higher than that of normal group (P <0.01). The cleavage of Desmin protein was obvious. The apoptosis rate of Tongyang Huoxue Recipe in high, medium and low dose groups was significantly lower than that in model group (P <0.01), and the average fluorescence intensity of Desmin was significantly higher than that in model group (P <0.01). Conclusion: The mechanism of Tongyang Huoxue Recipe for treating sick sinus syndrome may be related to combating the apoptosis of sinoatrial node cells and protecting Desmin morphological structure induced by ischemia-reperfusion.