Protection of gastrodin on PC12 cell injury induced by glutamate via inhibition of cell apoptosis

Abstract

Abstract: AIM: To study the protection and the molecular mechanisms of gastrodin on PC12 cells injury induced by glutamate. METHODS: The injury model was established by treating PC12 cells with glutamate in vitro and the cell activity was analyzed by MTT assay. Cell apoptosis morphological changes were detected by AO/EB staining with fluorescence microscope. The flow cytometry (FCM) was used to evaluate the level of intracellular reactive oxygen species (ROS) and to examine cell apoptosis rate via Annexin V-FITC/PI double staining. The expression of caspase-3 protein was determined by Western blot methods. RESULTS: It was indicated that gastrodin can apparently inhibit PC12 cell apoptosis induced by glutamate , it presented a dose-response relationship of the dosage 0.1-10 μmol/L. Gastrodin can also reduce ROS level, drop the expression of cleaved caspase-3 protein and decrease cell apoptosis rate, and gastrodin of 0.1-10 μmol/L presented a dose-response relationship. CONCLUSION: Gastrodin has a protective effect on apoptosis of PC12 cell within a certain dosage range, the protective mechanism of gastrodin on PC12 cell injury induced by glutamate may be related to gastrodin reducing the releasing of ROS, preventing occurrence of oxidative damage and thereby inhibiting caspase-3 pathway-dependent apoptosis in PC12 cell.

Key words: Gastrodin, Glutamate, PC12 cell, Apoptosis

CLC Number:

R965.2

WU Hai-yun, JIANG Gen-ling, LI Qing-lin. Protection of gastrodin on PC12 cell injury induced by glutamate via inhibition of cell apoptosis[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2012, 17(12): 1361-1367.

References

[1] Coyle JT, Puttfarcken P.Oxidative stress, glutamate, and neurodegenerative disorders[J]. Science, 1993, 262(5134): 689-695.
[2] Yang XD, Zhu J, Yang R, et al.Phenolic constituents from the rhizomes of Gastrodia elata[J]. Nat Prod Res, 2007, 21(2): 180-186.
[3] 张乐多, 龚晓健, 胡苗苗, 等. 天麻素抗血管性痴呆作用及其机理[J]. 中国天然药物, 2008, 6(2): 130-134.
[4] Zeng X, Zhang S, Zhang L, et al.A study of the neuroprotective effect of the phenolic glucoside gastrodin during cerebral ischemia in vivo and in vitro[J]. Planta Med, 2006, 72(15): 1359-1365.
[5] 柳星, 汪萌芽. 天麻素对海马内Aβ1-40注射阿尔兹海默病模型大鼠学习记忆的改善作用[J]. 中国临床药理学与治疗学, 2012, 17(4): 408-411.
[6] 王训翠, 朱国旗. 丹酚酸B对谷氨酸诱导的PC12细胞兴奋毒保护作用的研究[J]. 中国中药杂志, 2012, 37(3): 353-357.
[7] Westerink1 RHS, Ewing AG. The PC12 cell as model for neurosecretion[J]. Acta Physiol, 2008, 192(2): 273-285.
[8] Hawkins RA.The blood-brain barrier and glutamate[J]. Am J Clin Nutr, 2009, 90(3): 867-874.
[9] Arundine M, Tymianski M.Molecular mechanisms of glutamate-dependent neurodegeneration in ischemic and traumatic brain injury[J]. Cell Mol Life Sci, 2004, 61(6): 657-668.
[10] AnkarcronaM, Dypbukt JM, Bonfoco E, et al. Glutamate-induced neuronal death: a succession of necrosis or apoptosis depending on mitochondrial function[J]. Neuron, 1995, 15(4): 961-973.
[11] Penugonda S, Mare S, Goldstein G, et al.Effects of N-acetylcysteine amide (NACA),a novel thiol antioxidant against glutamate-induced cytotoxicity in neuronal cell line PC12[J]. Brain Res, 2005, 1056(2): 132-138.
[12] Lin MT, Beal MF.Mitochondrial dysfunction and oxidative stress in neurodegenerative diseases[J]. Nature, 2006, 443(19): 787-795.
[13] Emerit J, Edeas M, Bricaire F.Neurodegenerative diseases and oxidative stress[J]. Biomed Pharmacother, 2004, 58(1): 39-46.
[14] Hirata Y, Yamamoto H, Atta MS, et al.Chloroquine inhibits glutamate-induced death of a neuronal cell line by reducing reactive oxygen species through sigma-1 receptor[J]. J Neurochem, 2011, 119(4): 839-847.
[15] 邱瑜, 陈红专, 金正均. 谷氨酸神经细胞毒作用的新途径-谷氨酸/胱氨酸转运体介导机制[J]. 中国药理学通报, 2000, 16(3): 251-253.
[16] Wu G, Fang YZ, Yang S, et al.Glutathione metabolism and its implications for health[J]. J Nutr, 2004, 134(3): 489-492.
[17] Zheng TS, Hunot S, Kuida K, et al.Deficiency in caspase-9 or caspase-3 induces compensatory caspase activation[J]. Nat Med, 2000, 6(11): 1241-1247.
[18] Wang XC, Zhu GQ, Yang S, et al.Paeonol prevents excitotoxicity in rat pheochromocytoma PC12 cells via downregulation of ERK activation and inhibition of apoptosis[J]. Planta Med, 2011, 77(15): 1695-1701.