Gastrodin attenuates 6-hydroxydopamine-induced apoptosis in PC12 cells by modulating the PI3K signaling pathway and reducing reactive oxygen species levels

doi:10.12092/j.issn.1009-2501.2026.09.001

Abstract

Abstract:

AIM: To investigate the potential therapeutic mechanisms of Gastrodia elata and its main active component gastrodin (GSD) in Parkinson's disease (PD), focusing on apoptosis, oxidative stress, and the PI3K/Akt/mTOR signaling pathway. METHODS: A 6-hydroxydopamine (6-OHDA)-induced PC12 cell damage model was used to evaluate the effects of GSD on cell apoptosis, intracellular reactive oxygen species (ROS) levels, and the expression of PI3K/Akt/mTOR signaling pathway-related proteins in vitro. RESULTS: GSD significantly attenuated 6-OHDA-induced apoptosis in PC12 cells and modulated the activity of the PI3K signaling pathway. In addition, GSD effectively reduced intracellular ROS levels, and this effect was independent of PI3K pathway regulation. CONCLUSION: Gastrodin may exert neuroprotective effects by reducing intracellular ROS levels and modulating the PI3K/Akt/mTOR signaling pathway, thereby alleviating 6-OHDA-induced apoptosis in PC12 cells. These findings provide experimental evidence supporting the potential application of GSD in PD therapy.

Key words: Gastrodia elata Blume, Gastrodin, PI3K/Akt/mTOR signaling pathway, cell apoptosis

CLC Number:

Xinyu ZHANG, Shaowen WEN, Fei ZHAO, Lili WU, Jionglu ZHANG, Zehai FANG, Hongjie LIU. Gastrodin attenuates 6-hydroxydopamine-induced apoptosis in PC12 cells by modulating the PI3K signaling pathway and reducing reactive oxygen species levels[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2026, 31(9): 1153-1161.

Figures/Tables 5

Fig.1 Effect of gastrodin on apoptosis of PC12 cells A: cell viability of PC12 cells treated with different concentrations of 6-OHDA. B: cell viability of PC12 cells treated with different concentrations of gastrodin (GSD). C: effects of gastrodin at different concentrations on the viability of PC12 cells exposed to 60 μmol/L 6-OHDA. Cell viability in panels (A–C) was assessed using the MTT assay. D–E: flow cytometric analysis of apoptosis. F: Western blot analysis of Bax, BCL-2, and Caspase-3 protein expression in PC12 cells. G–I: quantitative analysis of Bax, BCL-2, and Caspase-3 protein levels. Protein bands were quantified by densitometric analysis and normalized to GAPDH. Data are presented as mean±SEM. For panels (A–C), n=5–6; for panels (F–I), n=3. *P<0.05, **P<0.01, ***P<0.001.

Fig.2 Effects of gastrodin on apoptosis of PC12 cells while adding LY294002 A–B: representative flow cytometric analysis of cell apoptosis. Data are presented as mean±SEM, n=3. ***P<0.001.

Fig.3 Effects of gastrodin on Bax, BCL-2 and Caspase-3 protein expression of PC12 cells while adding LY294002 A: Western blot analysis of Bax, BCL-2, and Caspase-3 protein expression in PC12 cells. B–D: quantitative analysis of Bax, BCL-2, and Caspase-3 protein levels. Protein bands were quantified by densitometric analysis and normalized to GAPDH. Data are presented as mean±SEM, n=3. **P<0.01, ***P<0.001.

Fig.4 Effect of gastrodin on the protein expression of PI3K, Akt and mTOR A: Western blot analysis of PI3K, Akt, and mTOR protein expression in PC12 cells. B–D: quantitative analysis of PI3K, Akt, and mTOR protein levels. Protein bands were quantified by densitometric analysis and normalized to GAPDH. E–F: representative flow cytometric fluorescence intensity profiles. Data are presented as mean±SEM, n=3. ***P<0.001.

Fig.5 The level of ROS production under the intervention of gastrodin A: Western blot analysis of PI3K, Akt, and mTOR protein expression in PC12 cells. B–D: quantitative analysis of PI3K, Akt, and mTOR protein levels. Protein bands were quantified by densitometric analysis and normalized to GAPDH. Data are presented as mean±SEM, n=3. *P<0.05, **P<0.01, ***P<0.001.

References 43

1	Abbas MM, Xu Z, Tan LCS. Epidemiology of Parkinson's disease-east versus west[J]. Mov Disord Clin Pract, 2018, 5 (1): 14- 28. doi: 10.1002/mdc3.12568
2	Collaborators GN. Global, regional, and national burden of neurological disorders, 1990-2016: a systematic analysis for the Global Burden of Disease Study 2016[J]. Lancet Neurol, 2019, 18 (5): 459- 480. doi: 10.1016/S1474-4422(18)30499-X
3	Narkiewicz J, Giachin G, Legname G. In vitro aggregation assays for the characterization of α-synuclein prion-like properties[J]. Prion, 2014, 8 (1): 19- 32. doi: 10.4161/pri.28125
4	Winklhofer KF, Haass C. Mitochondrial dysfunction in Parkinson's disease[J]. Biochim Biophys Acta, 2010, 1802 (1): 29- 44. doi: 10.1016/j.bbadis.2009.08.013
5	Wright R. Mitochondrial dysfunction and Parkinson's disease[J]. Nat Neurosci, 2022, 25 (1): 2.
6	刘娟, 李彦杰, 秦合伟, 等. 线粒体质量控制系统失调介导帕金森病的作用机制[J]. 实用医学杂志, 2024, 40 (11): 1479- 1482. doi: 10.3969/j.issn.1006-5725.2024.11.002
7	Meredith GE, Totterdell S, Potashkin JA, et al. Modeling PD pathogenesis in mice: advantages of a chronic MPTP protocol [J]. Parkinsonism Relat Disord, 2008, 14 Suppl 2(Suppl 2): S112-S115.
8	Kalyanaraman B, Cheng G, Hardy M. Gut microbiome, short-chain fatty acids, alpha-synuclein, neuroinflammation, and ROS/RNS: relevance to Parkinson's disease and therapeutic implications[J]. Redox Biol, 2024, 71, 103092. doi: 10.1016/j.redox.2024.103092
9	Schwienbacher C, Foco L, Picard A, et al. Plasma and white blood cells show different miRNA expression profiles in Parkinson's disease[J]. J Mol Neurosci, 2017, 62 (2): 244- 254. doi: 10.1007/s12031-017-0926-9
10	Poewe W, Seppi K, Tanner CM, et al. Parkinson disease[J]. Nat Rev Dis Primers, 2017, 3, 17013. doi: 10.1038/nrdp.2017.13
11	Guo QQ, Wang SS, Jiang XY, et al. Mitochondrial ROS triggers mitophagy through activating the DNA damage response signaling pathway [J]. Proc Natl Acad Sci U S A, 2025, 122(40): e2502841122.
12	Chou KL, Stacy M, Simuni T, et al. The spectrum of "off" in Parkinson's disease: What have we learned over 40 years ?[J]. Parkinsonism Relat Disord, 2018, 51, 9- 16. doi: 10.1016/j.parkreldis.2018.02.001
13	Zhang G, Xiong N, Zhang Z, et al. Effectiveness of traditional Chinese medicine as an adjunct therapy for Parkinson's disease: a systematic review and meta-analysis[J]. PLoS One, 2015, 10 (3): e0118498. doi: 10.1371/journal.pone.0118498
14	韩欣园, 刘翼骁, 肖依彤, 等. 中药多糖治疗帕金森病的研究进展[J]. 中华神经医学杂志, 2025, 24 (5): 524- 31. doi: 10.3760/cma.j.cn115354-20241226-00810
15	Leite Silva ABR, Gonçalves De Oliveira RW, Diógenes GP, et al. Premotor, nonmotor and motor symptoms of Parkinson's disease: a new clinical state of the art[J]. Ageing Res Rev, 2023, 84, 101834. doi: 10.1016/j.arr.2022.101834
16	姬之翼, 刘鸿高, 李杰庆, 等. 天麻化学成分、药理活性及质量标志物研究进展 [J]. 特产研究, 2025: 1-12.
17	中华人民共和国药典 2020版 [M]. 北京: 中国医药科技出版社, 2020.
18	杨飞, 王信, 马传江, 等. 天麻加工炮制、成分分析与体内代谢研究进展[J]. 中国中药杂志, 2018, 43 (11): 2207- 2215. doi: 10.19540/j.cnki.cjcmm.20180322.008
19	Xia C, Zhou H, Xu X, et al. Identification and investigation of miRNAs from gastrodia elata blume and their potential function[J]. Front Pharmacol, 2020, 11, 542405. doi: 10.3389/fphar.2020.542405
20	Dai Y, Ban W, Yang Z. Gastrodin, a promising natural small molecule for the treatment of central nervous system disorders, and its recent progress in synthesis, pharmacology and pharmacokinetics[J]. Int J Mol Sci, 2024, 25 (17): 9540. doi: 10.3390/ijms25179540
21	He X, Chen X, Yang Y, et al. The role of gastrodin in the management of CNS-related diseases: underlying mechanisms to therapeutic perspectives[J]. Phytother Res, 2024, 38 (11): 5107- 5133. doi: 10.1002/ptr.8314
22	Luo S, Kang SS, Wang ZH, et al. Akt phosphorylates NQO1 and triggers its degradation, abolishing its antioxidative activities in Parkinson's disease[J]. J Neurosci, 2019, 39 (37): 7291- 7305. doi: 10.1523/JNEUROSCI.0625-19.2019
23	Payne JM, Haebich KM, Mitchell R, et al. Brain volumes in genetic syndromes associated with mTOR dysregulation: a systematic review and meta-analysis[J]. Mol Psychiatry, 2025, 30 (4): 1676- 1688. doi: 10.1038/s41380-024-02863-4
24	张炅璐. 天麻活性成分治疗帕金森病作用机制的网络药理学预测及实验研究 [D]. 广州: 暨南大学, 2021.
25	Dorsey ER, Constantinescu R, Thompson JP, et al. Projected number of people with Parkinson disease in the most populous nations, 2005 through 2030[J]. Neurology, 2007, 68 (5): 384- 386. doi: 10.1212/01.wnl.0000247740.47667.03
26	Liu HJ, He TZ, Zhang JL, et al. [Systematic review and screening of basic Chinese herbs for traditional Chinese medicine compounds combined with levodopa medicine in treatment of Parkinson's disease][J]. Zhongguo Zhong Yao Za Zhi, 2020, 45 (24): 6043- 6052.
27	Chun YS, Kim J, Chung S, et al. Protective roles of monsonia angustifolia and its active compounds in experimental models of Alzheimer's disease[J]. J Agric Food Chem, 2017, 65 (15): 3133- 3140. doi: 10.1021/acs.jafc.6b04451
28	Ogut E, Armagan K, Gül Z. The role of syringic acid as a neuroprotective agent for neurodegenerative disorders and future expectations[J]. Metab Brain Dis, 2022, 37 (4): 859- 880. doi: 10.1007/s11011-022-00960-3
29	Kim IS, Choi DK, Jung HJ. Neuroprotective effects of vanillyl alcohol in Gastrodia elata Blume through suppression of oxidative stress and anti-apoptotic activity in toxin-induced dopaminergic MN9D cells[J]. Molecules, 2011, 16 (7): 5349- 5361. doi: 10.3390/molecules16075349
30	Hu D, Qing G, Liu X, et al. A study and in vitro evaluation of the bioactive compounds of broad bean sprouts for the treatment of Parkinson's syndrome[J]. Molecules, 2024, 29 (21): 5107. doi: 10.3390/molecules29215107
31	Xiao G, Tang R, Yang N, et al. Review on pharmacological effects of gastrodin[J]. Arch Pharm Res, 2023, 46 (9/10): 744- 770.
32	Guo H, Li C, Zhao J, et al. Mechanism of gastrodin against neurotoxicity based on network pharmacology, molecular docking and experimental verification[J]. Biomed Pharmacother, 2024, 180, 117611. doi: 10.1016/j.biopha.2024.117611
33	Zhan HD, Zhou HY, Sui YP, et al. The rhizome of Gastrodia elata Blume-An ethnopharmacological review[J]. J Ethnopharmacol, 2016, 189, 361- 385. doi: 10.1016/j.jep.2016.06.057
34	Liao W, Dong A, Hafeez F, et al. Celastrol protected the MPTP-injected mice Parkinson's disease model via redox regulation of CDC37[J]. Phytomedicine, 2025, 145, 157067. doi: 10.1016/j.phymed.2025.157067
35	Yuan J, Ofengeim D. A guide to cell death pathways[J]. Nat Rev Mol Cell Biol, 2024, 25 (5): 379- 395. doi: 10.1038/s41580-023-00689-6
36	Czabotar PE, Garcia-saez AJ. Mechanisms of BCL-2 family proteins in mitochondrial apoptosis[J]. Nat Rev Mol Cell Biol, 2023, 24 (10): 732- 748. doi: 10.1038/s41580-023-00629-4
37	Singh R, Letai A, Sarosiek K. Regulation of apoptosis in health and disease: the balancing act of BCL-2 family proteins[J]. Nat Rev Mol Cell Biol, 2019, 20 (3): 175- 193. doi: 10.1038/s41580-018-0089-8
38	Kaur S, Sehrawat A, Mastana SS, et al. Targeting calcium homeostasis and impaired inter-organelle crosstalk as a potential therapeutic approach in Parkinson's disease[J]. Life Sci, 2023, 330, 121995. doi: 10.1016/j.lfs.2023.121995
39	Nixon RA. Autophagy-lysosomal-associated neuronal death in neurodegenerative disease[J]. Acta Neuropathol, 2024, 148 (1): 42. doi: 10.1007/s00401-024-02799-7
40	Yu L, Wei J, Liu P. Attacking the PI3K/Akt/mTOR signaling pathway for targeted therapeutic treatment in human cancer[J]. Semin Cancer Biol, 2022, 85, 69- 94. doi: 10.1016/j.semcancer.2021.06.019
41	Goyal A, Agrawal A, Verma A, et al. The PI3K-AKT pathway: a plausible therapeutic target in Parkinson's disease[J]. Exp Mol Pathol, 2023, 129, 104846. doi: 10.1016/j.yexmp.2022.104846
42	Yang S, Lian G. ROS and diseases: role in metabolism and energy supply[J]. Mol Cell Biochem, 2020, 467 (1/2): 1- 12. doi: 10.1007/s11010-019-03667-9
43	Song J, Wang Y, Yuan X, et al. Stretching magnitude-dependent inactivation of AKT by ROS led to enhanced p53 mitochondrial translocation and myoblast apoptosis[J]. Mol Biol Cell, 2019, 30 (10): 1182- 1197. doi: 10.1091/mbc.E18-12-0770

[1]	ZHANG Jianxin, YANG Xiaolai. Glycyrrhizin inhibits the neurotoxic effects and mechanisms induced by morphine [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2024, 29(5): 554-560.
[2]	LIU Min, DING Yanxia, ZHANG Yegui, CHAN Cuicui, GONG Rujie, NI Jingzhong. Effects of gastrodin on the expression of BDNF and IL-6 in the striatum of rats with cerebral ischemia [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2024, 29(4): 440-446.
[3]	HUANG Xiaomei, ZHI Hui, CHEN Hao, LU Linming, ZHU Xiaoqun, WANG Lizhen, ZHOU Jue, PANG Jinjin, XU Jinliang. Effects of the proliferation, migration and apoptosis of AHVAC-Ⅰ on gastric cancer MKN-28 cells [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2024, 29(3): 270-276.
[4]	ZHANG Xuejiao, LIU Jieting, LI Luxin, CHEN Peijian, DING Minglu, SUN Mengwei, CHU Yanhui, ZHANG Zhen . Effects and mechanism of dapagliflozin on myocardial injury in type 1 diabetes mice [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2023, 28(3): 257-265.
[5]	WANG Youlin, LI Shanshan, WANG Meng, GUO Yuting, CHENG Hui, TIAN Shasha, LI Qinglin. Gastrodin inhibits H₂O₂-induced ferroptosis of PC12 cells [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2021, 26(5): 532-538.
[6]	WU Wei, LI Guangpeng, ZHANG Jiangbo, KONG Lingyu, CAI Junyan, YANG Feiyun. Gastrodin combined with dexamethasone protects H9C2 cell from injury induced by oxygen-glucose deprivation [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2021, 26(11): 1244-1249.
[7]	WENG Dandan, WANG Junbo, LI Guangxiao. Mechanism of PI3K/AKT/mTOR signaling pathway on apoptosis of hepatocyte induced by GCDC [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2019, 24(2): 121-127.
[8]	. Effects and mechanisms of nicotinic acid curcumin on apoptosis induced by low shear stress in human umbilical vein endothelial cells [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2018, 23(8): 855-861.
[9]	MA Chenghua, ZHOU Haijing, SHI Wen, WANG Zhiqiang, YANG Lan, CHEN Ping, ZHENG Xueli, MA Rui, MA Lijuan. Mechanism of apoptosis of mucoepidermoid carcinoma MEC-1 cells induced by ethanol extract of Lamiophlomis rotate [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2017, 22(3): 253-259.
[10]	LUO Ziyin, XIANG Bingqian, GAO Hui, QIU Xiaoxiao, QIAN Xiaoying, WANG Wantie. Effects of dexmedetomidine on JNK in A549 cells with hypoxia/reoxygenation injury [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2017, 22(12): 1346-1351.
[11]	SHAO Mei-qin, CHEN Hai-e, HE Jin-bo, MA Ying-chun, HUANG Lin-jing, CHEN Dan, WANG Yang, WANG Wan-tie. Effects of JNK on ischemic postconditioning attenuating pneumocyte apoptosis after lung ischemia/reperfusion injury [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2015, 20(5): 531-536.
[12]	ZHOU Jie, YAN Ling, HUANG Duo-xin. Mfn2 gene transfer inhibites neonatal rat cardiomyocytes apoptosis induced by ischemia/reperfusion via activation of the Ras-PI3K-Akt signaling pathway [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2014, 19(9): 973-978.
[13]	GONG Deng-hui, ZHENG Wei-hong, LUO Ni, TANG Gui-chen. Effects and the mechanism of gastrodin on chemotherapy-induced neuropathic pain through the expression of Iba1 of spinal dorsal horn [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2014, 19(7): 743-746.
[14]	WANG Ming, ZHANG Yao, XIE Xiang-rong, QI Zhi-lin, BI Fu-yong. Oridonin induces MDA-MB-231 cells apoptosis through PI3K/Akt pathway in vitro [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2013, 18(2): 161-165.
[15]	LIU Xing, WANG Meng-ya. Improving effects of gastrodin on learning and memory in model rats with Alzheimer disease induced by intra-hippocampal Aβ_1-40 injection [J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2012, 17(4): 408-411.