梓醇对PC12细胞氧糖剥夺损伤的保护作用及对电压依赖性钾通道亚型表达的影响  

The protective effect of catalpol on injured PC12 cells induced by oxygen and sugar deprivation and the influence on voltage dependent potassium channel subtypes expression

作　　者：张晓双[1] 胡锐[1] 刘继平[1] 孙建宁[2] 

机构地区：[1]陕西中医药大学,咸阳712046 [2]北京中医药大学中药学院,北京100102

出　　处：《中药药理与临床》2017年第5期31-34,共4页Pharmacology and Clinics of Chinese Materia Medica

基　　金：陕西省教育厅自然科学专项(15JK1216)

摘　　要：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 examine the protective effect of catalpol on injured PC12 cells induced by oxygen and sugar deprivation and the influence on voltage dependent potassium channel subtypes expression. Methods： The oxygen and sugar deprivation model of PC12 cells was induced by sodium dithionite（Na2 S2 O4） combined with sugar-free culture medium,catalpol was given 24 hours before oxygen and sugar deprivation,the cell survival was determined by MTT; the cell morphology was observed by the inverted microscope; lactate dehydrogenase（LDH） activity of cell cultures was determined; intracellular superoxide dismutase（SOD） activity and malondialdehyde（MDA） content,reduced glutathione（GSH） content were determined; The expression of voltage dependent potassium channels Kv1. 4,Kv1. 5,Kv2. 1 and Kv4. 2 protein in PC12 cells were determined by western blot. Results： The cell survival rate of injured PC12 cells induced by oxygen and glucose deprivation decreased,100 mu mol/L,10 mu mol/Lof catalpol were obviously against oxygen glucose deprivation damage in PC12 cells,the cell survival rate significantly increased. After PC12 cells received oxygen glucose deprivation damage,the release of LDH in medium increased,SOD activity decreased,MDA content increased,GSH content significantly decreased in cells,catalpol was against the oxidative stress,inhibited LDH release,increased SOD activity,GSH content,reduced the MDA content in cells,removed excess free radicals,catalpol had certain protective effect on injured PC12 cell induced by oxygen and glucose deprivation. After oxygen and glucose deprivation in PC12 cells,voltage dependent potassium channels Kv1. 5 and Kv2. 1 protein expressions significantly rose,catalpol decreased Kv1. 5 and Kv2. 1 protein expressions in PC12 cells after oxygen glucose deprivation damage. Conclusion： Catalpol has the protective efffect on PC12 cells after oxygen glucose deprivation damage,has resistance to oxidative stress damage,decrease voltage dependent potassium channels Kv1. 5 a

关 键 词：梓醇 PC12 电压依赖性钾通道 氧糖剥夺 

分 类 号：R285[医药卫生—中药学]