葛根素保护双氧水诱导的SH-SY5Y细胞凋亡  

Effects of puerarin on H_2O_2-induced SH-SY5Y cell apoptosis

作　　者：陈龙菊 吴建清[2] 吴太鼎[2] 陈传奇 谭云霞[2] 柯尊记 

机构地区：[1]上海中医药大学基础医学院生化教研室,上海201203 [2]湖北民族学院医学院解剖教研室,湖北恩施445000

出　　处：《中国药理学通报》2018年第3期343-347,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81260192);湖北省教育厅科学研究重点项目(No D20151903)

摘　　要：目的探讨葛根素对双氧水(H_2O_2)诱导的SH-SY5Y细胞凋亡的保护作用及其机制。方法建立体外神经元损伤模型,MTT法观察细胞存活率;Hoechst 33342染色观察细胞核改变;JC-1染色检测细胞线粒体膜电位的改变;酶活性检测线粒体caspase-3和caspase-9的变化;Western blot检测细胞中Bcl-2、Bax、p-Akt、Akt蛋白的表达。结果与H_2O_2模型组相比,葛根素预处理能明显改善H_2O_2诱导的SHSY5Y细胞存活率下降(P<0.05),缓解H_2O_2引起的线粒体膜电位的下降(P<0.01),抑制caspase-3和caspase-9的酶活性(P<0.01),减少H_2O_2诱导的细胞凋亡。此外,葛根素还促进细胞内p-Akt、Bcl-2蛋白表达,抑制Bax蛋白表达,而这种作用能被PI3K/Akt的抑制剂LY294002所抑制。结论葛根素可保护H_2O_2诱导的SH-SY5Y细胞凋亡,这种保护作用可能是通过激活PI3K/Akt信号通路实现的。Aim To investigate the neuroprotective effects of puerarin on H_2O_2-induced SH-SY5 Y cell apoptosis and the molecular mechanisms underlying the neuroprotective effects. Methods Neuron injury model was established in vitro through H_2O_2-induced SHSY5 Y injury. MTT assay was performed to detect the effect of puerarin on H_2O_2-induced SH-SY5 Y survival rates. Hoechst 33342 staining was used to observe the cell apoptosis. JC-1 staining was employed to detect the level of mitochondria membrane poential. Caspase-3 was determined by caspase-3 catalyze the substrate specificity Ac-DEVD-p NA. Caspase-9 was determined by caspase-9 catalyze the substrate specificity Ac-LEHD-p NA. The effects of puerarin on the protein level of Bcl-2,Bax,p-Akt and Akt were determined by Western blot. Results The cell survival rate significantly increased after puerarin pretreatment compared with H_2O_2 model group. Furthermore, puerarin pretreatment not only inhibited the decreasing of mitochondrial membrane potential,increasing of caspase-3,caspase-9 enzymatic activity and the expression of Bax,but also promoted the expression of p-Akt and Bcl-2,which was prevented by LY294002,an inhibitor of PI3 K/Akt. Conclusion Puerarin can play a neuroprotective role for SH-SY5 Y cell apoptosis induced by H_2O_2,maybe via activating PI3 K/Akt signaling pathway.

关 键 词：葛根素 过氧化氢 人神经母细胞瘤 凋亡 PI3K AKT 

分 类 号：R282.71[医药卫生—中药学] R322.8[医药卫生—中医学]