Epigallocatechin-3-gallate suppresses 1-methyl-4-phenyl-pyridine-induced oxidative stress in PC12 cells via the SIRT1/PGC-1α signaling pathway

Department of Neurology, Fujian Institute of Geriatrics, The Affiliated Union Hospital of Fujian Medical University, Fuzhou, China.
BMC Complementary and Alternative Medicine (Impact Factor: 1.88). 06/2012; 12:82. DOI: 10.1186/1472-6882-12-82
Source: PubMed

ABSTRACT Parkinson's disease is a high incidence neurodegenerative disease in elderly people, and oxidative stress plays an important role in the pathogenesis. Oxygen metabolism in the brain is high, which lacks an antioxidative protection mechanism. Recently, it has been found that polyphenols play an important role in antioxidation. (-)-epigallocatechin-3-gallate (EGCG) is an important component of tea polyphenols and its biological effects, such as strong antioxidation, scavenging of free radicals and anti-apoptosis, can pass through the blood brain barrier. The SIRT1/PGC-1α signaling pathway has not been reported in PC12 cells. Therefore, research of the protective mechanism of EGCG in PC12 cells damaged by -methyl-4-phenyl-pyridine (MMP+) may provide a new insight into protect against and treatment of Parkinson's disease.
MPP(+)-treated highly differentiated PC12 cells were used as the in vitro cell model. An MTT assay was used to investigate cell viability after EGCG treatment, a dichlorofluorescin diacetate assay was used to measure reactive oxygen species (ROS) production, western blot analysis was used to observe PGC-1α and SIRT1 protein expression, and real-time PCR to observe PGC-1α, SOD1 and GPX1 mRNA expression.
PC12 cell viability was significantly reduced after MPP(+) treatment by 11.46% compared with that of the control (P < 0.05). However, cell viability was unchanged by 10 μmol/L EGCG treatment. In co-treatments with EGCG and MPP(+), cell viability was significantly increased by 12.92% (P < 0.05) and MPP(+)-induced ROS production was markedly decreased. PGC-1α mRNA expression was obviously upregulated by 21.51% (P < 0.05), and SOD1 and GPX1 mRNA expression was slightly increased by 12.94% and 15.63% (P > 0.05), respectively, by treatment with EGCG and then MPP(+) for 12 h. The mRNA expression of PGC-1α, SOD1 and GPX1 was increased by 25.17%, 40% and 146% (all P < 0.05), respectively, by treatment with EGCG and then MPP(+) for 24 h. Such effects were not observed with MPP(+) treatment alone.
The SIRT1/PGC-1α pathway is one of the mechanisms of EGCG suppression of MPP(+)-induced injury of PC12 cells.

  • [Show abstract] [Hide abstract]
    ABSTRACT: The molecular mechanism responsible for degenerative process in the nigrostriatal dopaminergic system in Parkinson's disease (PD) remains unknown. One major advance in this field has been the discovery of several genes associated to familial PD, including alpha synuclein, parkin, LRRK2, etc., thereby providing important insight toward basic research approaches. There is an consensus in neurodegenerative research that mitochon dria dysfunction, protein degradation dysfunction, aggregation of alpha synuclein to neurotoxic oligomers, oxidative and endoplasmic reticulum stress, and neuroinflammation are involved in degeneration of the neuromelanin-containing dopaminergic neurons that are lost in the disease. An update of the mechanisms relating to neurotoxins that are used to produce preclinical models of Parkinson´s disease is presented. 6-Hydroxydopamine, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, and rotenone have been the most wisely used neurotoxins to delve into mechanisms involved in the loss of dopaminergic neurons containing neuromelanin. Neurotoxins generated from dopamine oxidation during neuromelanin formation are likewise reviewed, as this pathway replicates neurotoxin-induced cellular oxidative stress, inactivation of key proteins related to mitochondria and protein degradation dysfunction, and formation of neurotoxic aggregates of alpha synuclein. This survey of neurotoxin modeling-highlighting newer technologies and implicating a variety of processes and pathways related to mechanisms attending PD-is focused on research studies from 2012 to 2014.
    Neurotoxicity Research 01/2015; 27(3). DOI:10.1007/s12640-015-9519-y · 3.15 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Reports suggest that silent information regulation 2 homolog 3 (SIRT3) protects cardiomyocytes from oxidative stress-mediated death. SIRT3, a mitochondrial protein, is an essential regulator of mitochondrial function, and this regulation is important in many cerebrovascular diseases, especially stroke. Here, we investigated the role of SIRT3 in ischemia-induced neuronal death due to oxygen-glucose deprivation (OGD) using an in vitro model of cerebral ischemia. We found that exposure of differentiated PC12 cells to OGD for 6 h caused a marked decrease in cell viability and up regulated SIRT3. SIRT3 knockdown using short interfering RNA (siRNA) exacerbated OGD-induced injury whereas application of recombinant SIRT3 protected against OGD-induced cell death. Pre-treatment of the cells in which the SIRT3 gene was knocked down with recombinant SIRT3 before OGD partially restored cell viability and concomitantly reduced lactate dehydrogenase (LDH) release and increased ATP generation in mitochondria. Recombinant SIRT3 treatment resulted in increased expression of PGC-1α and manganese superoxide dismutase (MnSOD). After knockdown of PGC-1α or MnSOD, recombinant SIRT3 failed to protect against OGD-induced injury. We also found that the protein and mRNA expression of PGC-1α was down regulated following SIRT3 knockdown. The expression level of SIRT3 was reduced when the PGC-1α gene was knocked down. Both SIRT3 and PGC-1α knockdown led to reduced mitochondrial membrane potential (△ψ) and Ca(2+) transients, especially under OGD conditions. Thus, our data suggest that SIRT3 protects PC12 cells from hypoxic injury via a mechanism that may involve PGC-1α and MnSOD. SIRT3 and PGC-1α regulate each other under physiologic and OGD conditions, thereby partially protecting against hypoxia or ischemia. Copyright © 2014. Published by Elsevier Ltd.
    Neuroscience 11/2014; 286. DOI:10.1016/j.neuroscience.2014.11.045 · 3.33 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Background: Previous studies have suggested that high doses of (-)-epigallocatechin-3-gallate (EGCG) can induce toxicity in the liver, kidneys, and intestine. However, there have been no reports of myocardiotoxicity following treatment with EGCG. In this study, we investiged the proliferation and apoptosis of H9C2 cardiomyocytes treated with high dose of EGCG. Methods: Cell proliferation was measured by CCK8 assay, cell apoptosis rate was evaluated by TUNEL assay, and the expression alterations of Sirtuin 1 (SIRT1) protein was detected by Western blotting. Results: EGCG inhibits proliferation and induces apoptosis in time- and dose-dependent manner in H9C2 cardiomyocytes. SIRT1 participates in the inhibitory effect of EGCG on cell proliferation and apoptosis induction in H9C2 cardiomyocytes. Conclusion: This study demonstrates that high doses of EGCG inhibit proliferation and induce apoptosis in H9C2 cardiomyocytes. Down-regulation of SIRT 1 protein expression may be involved.
    Pharmazie 01/2015; 70(1). DOI:10.1691/ph.2015.4717 · 1.00 Impact Factor

Full-text (3 Sources)

Available from
Jun 10, 2014