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目的:考察梓醇对氧糖剥夺的PC12细胞损伤的保护作用及对电压依赖性钾通道亚型表达的影响。方法:采用连二亚硫酸钠(Na2S2O4)合并无糖培养基造成PC12细胞氧糖剥夺模型,给予梓醇预处理24h后,MTT法检测细胞存活率;倒置显微镜观察细胞形态;测定细胞培养液乳酸脱氢酶(LDH)活性;测定细胞内超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量;还原型谷胱甘肽(GSH)含量;Western blot检测PC12细胞内电压依赖性钾通道Kv1.4、Kv1.5、Kv 2.1、Kv 4.2蛋白的表达。结果:PC12细胞氧糖剥夺损伤后细胞存活率下降,梓醇100μmol/L、10μmol/L能明显对抗氧糖剥夺对PC12细胞的损伤,使细胞存活率明显增加。PC12细胞氧糖剥夺损伤后培养液中LDH释放增多,细胞内SOD活性显著下降,MDA含量升高、GSH含量降低,梓醇能抗氧化应激,抑制细胞LDH释放,升高细胞内SOD活力,升高细胞内GSH含量,降低细胞内MDA含量。PC12细胞氧糖剥夺损伤后,细胞内电压依赖性钾通道Kv1.5和Kv2.1蛋白表达明显上调,梓醇能下调氧糖剥夺PC12细胞内Kv1.5和Kv2.1蛋白表达。结论:梓醇对缺糖缺氧PC12有保护作用,能抗氧化应激损伤,其机制可能与下调电压依赖性钾通道Kv1.5和Kv2.1蛋白表达有关。
Objective: To investigate the protective effect of catalpol on the injury induced by oxygen sugar deprivation in PC12 cells and its effect on the expression of voltage-dependent potassium channel subtypes. Methods: Oxygen-glucose deprivation model of PC12 cells was induced by sodium dithionite (Na2S2O4) combined with sugar-free medium. After being pretreated with catalpol for 24 h, cell viability was determined by MTT assay. Cell morphology was observed by inverted microscope. (LDH) activity was measured. The activity of superoxide dismutase (SOD), content of malondialdehyde (MDA) and content of reduced glutathione (GSH) in cells were measured. The voltage-dependent potassium channel Kv1 .4, Kv1.5, Kv 2.1, Kv 4.2 protein expression. Results: The survival rate of PC12 cells was decreased after oxygen-glucose deprivation injury. Catalpol 100μmol / L and 10μmol / L could significantly antagonize the injury of PC12 cells induced by oxy-glucose deprivation and significantly increase the cell survival rate. In the culture of PC12 cells, the release of LDH increased, the activity of SOD decreased significantly, the content of MDA increased, the content of GSH decreased. Catalpol can resist the oxidative stress, inhibit the release of LDH, increase the activity of SOD, Increase intracellular GSH content, reduce intracellular MDA content. After oxygen-glucose deprivation injury of PC12 cells, the expression of Kv1.5 and Kv2.1 in Kv1.5 and Kv2.1 in PC12 cells were significantly up-regulated. Catalpol could down-regulate the expression of Kv1.5 and Kv2.1 in PC12 cells. CONCLUSION: Catalpol can protect against hypoxia and hypoxia-induced PC12 and protect against oxidative stress. The mechanism may be related to the down-regulation of Kv1.5 and Kv2.1 protein expression in voltage-dependent potassium channels.