神经药理学报 ›› 2017, Vol. 7 ›› Issue (5): 1-9.DOI: 10.3969/j.issn.2095-1396.2017.05.001

• 研究论文 •    下一篇

黄芪甲苷基于PI3K/Akt/Bcl-2 信号通路对PC12 细胞OGD/R 损伤的保护机制研究

张佳琳,李元元,杨文亮,刘富甲,田子夏,谢欣梅,庞晓斌   

  1. 1. 河南大学药物研究所,开封,475004,中国
    2. 河南应用技术职业学院医学院,开封,475004,中国
  • 出版日期:2017-10-26 发布日期:2017-12-01
  • 通讯作者: 庞晓斌,E-mail:hndxpxb@163.com

Astragaloside IV Protect PC12 Cells under Stress Induced by Oxygen-Glucose Deprivation Involving PI3K/Akt/Bcl-2 Pathway

ZHNAG Jia-lin1,LI Yuan-yuan2,YANG Wen-liang1,LIU Fu-jia1,TIAN Zi-xia1,XIE Xin-mei1,PANG Xiao-bin1   

  1. 1. Pharmaceutical Institute,Henan University,Kaifeng,475004,China
    2. College of Medicine,College of Applied Technology of Henan,Kaifeng,475004,China
  • Online:2017-10-26 Published:2017-12-01
  • Contact: 庞晓斌,E-mail:hndxpxb@163.com

摘要:

目的:探究黄芪甲苷(astragaloside IV,AS-IV)对大鼠嗜铬细胞瘤(pheochromocytoma 12,PC12)细胞缺氧缺糖复供(oxygen deprivation reoxygenation,OGD/R)损伤的保护作用及基于磷脂酰肌醇3 激酶(phosphatidylinositol 3 kinase,PI3K)/ 蛋白激酶B(protein kinase B,PKB,又称Akt)及B 细胞淋巴瘤-2 蛋白(B-cell lymphoma-2,Bcl-2) PI3K/Akt/Bcl-2 信号通路的机制研究。方法:体外培养 PC12 细胞,细胞随机分为空白对照组、模型组(OGD/R),尼莫地平阳性对照组(5 μmol·L-1)和黄芪甲苷组(1.00、0.10、0.01 μmol·L-1),除空白组外其余各组均建立体外PC12 细胞OGD/R 模型。CCK-8 试剂盒法检测PC12 细胞存活率;乳酸脱氢酶(lactate dehydrogenase,LDH)试剂盒法测定乳酸脱氢酶的漏出量;采用Hoechst33342 和碘化丙啶(propidium iodide,PI) 双染法检测各组细胞凋亡与坏死情况;Western blot 检测各组细胞Akt、p-Akt、Bcl-2、Bcl-2 相关 X 蛋白(Bcl-2 associated X protein,Bax)和含半胱氨酸的天冬氨酸蛋白水解酶-3 (cysteinyl aspartate specific proteinase-3,Caspase-3)的蛋白表达情况。结果:黄芪甲苷提高PC12 细胞OGD/R 损伤的细胞存活率,降低LDH 漏出量和细胞凋亡。黄芪甲苷浓度依赖性上调p-Akt/Akt、Bcl-2 的蛋白表达和下调促凋亡蛋白Bax、Caspase-3 的表达。PI3K/Akt 通路的抑制剂LY2940002 能抑制黄芪甲苷对OGD/R 损伤的PC12 细胞中凋亡相关蛋白的影响。结论:黄芪甲苷对OGD/R 损伤的PC12 细胞具有保护作用,且其保护作用机制可能与PI3K/Akt/Bcl-2 信号通路有关。

关键词: 黄芪甲苷, PC12, OGD/R, PI3K/Akt/Bcl-2 信号通路, 神经保护

Abstract:

Objective:To investigate the protective effect of astragaloside IV (AS-IV) on oxygen and glucose deprivation/reoxygenation of PC12 cells and the mechanism of PI3K/Akt/Bcl-2 signaling pathway. PC12 cells were cultured in vitro and nimodipine was used as a positive control. Pretreatment was carried out with different concentrations of AS-IV(1.00、0.10、0.01 μmol·L-1). Methods:The oxygen-glucose deprivasion/reperfution (OGD/R) model was made. CCK-8 assay was used to detect the viability of PC12 cells. LDH kit was used to measure the leakage of lactate dehydrogenase. Hoechst 33342/PI staining was used to detect the apoptosis and necrosis. The expression of Akt,p-Akt,Bcl-2,Bax,and Caspase-3 were detected by western blot. Results:The results showed that AS-IV can increase the cell viability of OGD/R in PC12 cells and decreasethe leakage rate of LDH and apoptosis. The expression of p-Akt/Akt and Bcl-2 protein were up-regulated and the expression of pro-apoptotic protein Bax and Caspase-3 were down-regulated in a dose-dependent manner. PI3K/Akt signaling pathway inhibitor LY2940002 inhibits the effects of AS-IV on apoptosis-related proteins in OGD/R injury of PC12 cells. Conclusion:AS-IV has protective effect on PC12 cells injured by OGD/R,and the protective mechanism is related to PI3K/Akt/Bcl-2 signaling pathway.

Key words: astragaloside IV(AS-IV), PC12;OGD/R, PI3K/Akt/Bcl-2 signaling pathway, neuroprotection

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