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Hydroxysafflor Yellow A Protects PC12 Cells Against the Apoptosis Induced by Oxygen and Glucose Deprivation

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Abstract

Hydroxysafflor yellow A (HSYA) was reported neuroprotective under several ischemic models in vivo. In this study, the direct effect of HSYA against oxygen–glucose deprivation (OGD) inducing acute neuronal injury and the underling mechanisms in vitro were investigated. Four-hour oxygen and glucose deprivation (OGD) followed by 20 h reperfusion (adding back oxygen and glucose, OGD-R) was used to induce in vitro ischemia reperfusion injury in differentiated rat pheochromocytoma PC12 cells. HSYA (1, 10, and 100 μmol/l) was added to the cultures 30 min prior to the ischemic insult and was present during OGD and reoxygenation phases. The survival rate of PC12 cells was detected by MTT assay. The contents of malondialdehyde (MDA), superoxide dismutase (SOD) activity were elevated by biochemical method. Hoechst 33258 staining and flow cytometric analysis were used to detect apoptosis; western blotting was used to detect the expression of Bcl-2, Bax, and Cytochrome C protein. The activity of caspase-3 was assessed by colorimetry. HSYA concentration-dependently attenuated neuronal damage with characteristics of increasing injured neuronal absorbance of MTT, decreasing cell apoptosis, and antagonizing decreases in SOD activity and increase in MDA level induced by OGD-R. Moreover, the down-regulation of Bcl-2, up-regulation of Bax and the release of mitochondrial cytochrome c to cytosol and the consequent activation of caspase-3 were reversed by HSYA in a dose-dependent manner. These results suggest that apoptosis is an important characteristic of OGD-R-induced PC 12 death and that treatment of PC12 cells with HSYA can block OGD-R-induced apoptosis through suppression of intracellular oxidative stress and mitochondria dependent caspase cascade.

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References

  • Almeida A, Delgado-Esteban M, Bolanos JP, Medina JM (2002) Oxygen and glucose deprivation induces mitochondrial dysfunction and oxidative stress in neurones but not in astrocytes in primary culture. J Neurochem 81(2):207–217

    Article  PubMed  CAS  Google Scholar 

  • Brongholi K, Souza DG, Bainy AC, Dafre AL, Tasca CI (2006) Oxygen–glucose deprivation decreases glutathione levels and glutamate uptake in rat hippocampal slices. Brain Res 1083(1):211–218

    Article  PubMed  CAS  Google Scholar 

  • Broughton BR, Reutens DC, Sobey CG (2009) Apoptotic mechanisms after cerebral ischemia. Stroke 40(5):e331–e339

    Article  PubMed  Google Scholar 

  • Brunelle JK, Letai A (2009) Control of mitochondrial apoptosis by the Bcl-2 family. J Cell Sci 122(Pt 4):437–441

    Article  PubMed  CAS  Google Scholar 

  • Fan LH, Wang KZ, Cheng B, Wang CS, Dang XQ (2006) Anti-apoptotic and neuroprotective effects of tetramethylpyrazine following spinal cord ischemia in rabbits. BMC Neurosci 7:48

    Article  PubMed  Google Scholar 

  • Fiskum G, Rosenthal RE, Vereczki V, Martin E, Hoffman GE, Chinopoulos C, Kowaltowski A (2004) Protection against ischemic brain injury by inhibition of mitochondrial oxidative stress. J Bioenerg Biomembr 36(4):347–352

    Article  PubMed  CAS  Google Scholar 

  • He H, Yang X, Shi M, Zeng X, Yang J, Wu L, Li L (2008) Protective effects of hydroxysafflor yellow A on acute and chronic congestive cardiac failure mediated by reducing ET-1, NOS and oxidative stress in rats. J Pharm Pharmacol 60(1):115–123

    Article  PubMed  CAS  Google Scholar 

  • Ji DB, Zhu MC, Zhu B, Zhu YZ, Li CL, Ye J, Zhu HB (2008) Hydroxysafflor yellow A enhances survival of vascular endothelial cells under hypoxia via upregulation of the HIF-1 alpha-VEGF pathway and regulation of Bcl-2/Bax. J Cardiovasc Pharmacol 52(2):191–202

    Article  PubMed  CAS  Google Scholar 

  • Ji DB, Zhang LY, Li CL, Ye J, Zhu HB (2009) Effect of hydroxysafflor yellow A on human umbilical vein endothelial cells under hypoxia. Vascul Pharmacol 50(3–4):137–145

    Article  PubMed  CAS  Google Scholar 

  • Jiang H, Koubi D, Zhang L, Kuo J, Rodriguez AI, Hunter TJ, Gautam SC, Levine RA (2005) Inhibitors of iNOS protects PC12 cells against the apoptosis induced by oxygen and glucose deprivation. Neurosci Lett 375(1):59–63

    Article  PubMed  CAS  Google Scholar 

  • Krantic S, Mechawar N, Reix S, Quirion R (2007) Apoptosis-inducing factor: a matter of neuron life and death. Prog Neurobiol 81(3):179–196

    Article  PubMed  CAS  Google Scholar 

  • Leibowitz B, Yu J (2010) Mitochondrial signaling in cell death via the Bcl-2 family. Cancer Biol Ther 9(6):417–422

    Article  PubMed  CAS  Google Scholar 

  • Marzocco S, Popolo A, Bianco G, Pinto A, Autore G (2010) Pro-apoptotic effect of methylguanidine on hydrogen peroxide-treated rat glioma cell line. Neurochem Int 57(5):518–524

    Article  PubMed  CAS  Google Scholar 

  • Meloni BP, Majda BT, Knuckey NW (2001) Establishment of neuronal in vitro models of ischemia in 96-well microtiter strip-plates that result in acute, progressive and delayed neuronal death. Neuroscience 108(1):17–26

    Article  PubMed  CAS  Google Scholar 

  • Niizuma K, Endo H, Chan PH (2009) Oxidative stress and mitochondrial dysfunction as determinants of ischemic neuronal death and survival. J Neurochem 109(Suppl 1):133–138

    Article  PubMed  CAS  Google Scholar 

  • Nilsen J, Chen S, Irwin RW, Iwamoto S, Brinton RD (2006) Estrogen protects neuronal cells from amyloid beta-induced apoptosis via regulation of mitochondrial proteins and function. BMC Neurosci 7:74

    Article  PubMed  Google Scholar 

  • Patten DA, Germain M, Kelly MA, Slack RS (2010) Reactive oxygen species: stuck in the middle of neurodegeneration. J Alzheimers Dis 20(Suppl 2):S357–S367

    PubMed  Google Scholar 

  • Rawal A, Muddeshwar M, Biswas S (2004) Effect of Rubia cordifolia, Fagonia cretica linn, and Tinospora cordifolia on free radical generation and lipid peroxidation during oxygen-glucose deprivation in rat hippocampal slices. Biochem Biophys Res Commun 324(2):588–596

    Article  PubMed  CAS  Google Scholar 

  • Shan LQ, Ma S, Qiu XC, Zhou Y, Zhang Y, Zheng LH, Ren PC, Wang YC, Fan QY, Ma BA (2010) Hydroxysafflor yellow A protects spinal cords from ischemia/reperfusion injury in rabbits. BMC Neurosci 11:98

    Article  PubMed  Google Scholar 

  • Shen CC, Huang HM, Ou HC, Chen HL, Chen WC, Jeng KC (2004) Protective effect of nicotinamide on neuronal cells under oxygen and glucose deprivation and hypoxia/reoxygenation. J Biomed Sci 11(4):472–481

    Article  PubMed  CAS  Google Scholar 

  • Singh M, Sharma H, Singh N (2007) Hydrogen peroxide induces apoptosis in HeLa cells through mitochondrial pathway. Mitochondrion 7(6):367–373

    Article  PubMed  CAS  Google Scholar 

  • Susnow N, Zeng L, Margineantu D, Hockenbery DM (2009) Bcl-2 family proteins as regulators of oxidative stress. Semin Cancer Bio l19:42–49

    Article  Google Scholar 

  • Tabakman R, Jiang H, Shahar I, Arien-Zakay H, Levine RA, Lazarovici P (2005) Neuroprotection by NGF in the PC12 in vitro OGD model: involvement of mitogen-activated protein kinases and gene expression. Ann N Y Acad Sci 1053:84–96

    Article  PubMed  CAS  Google Scholar 

  • Tian J, Li G, Liu Z, Fu F (2008) Hydroxysafflor yellow A inhibits rat brain mitochondrial permeability transition pores by a free radical scavenging action. Pharmacology 82(2):121–126

    Article  PubMed  CAS  Google Scholar 

  • Turrens JF (2003) Mitochondrial formation of reactive oxygen species. J Physiol 552(Pt 2):335–344

    Article  PubMed  CAS  Google Scholar 

  • Wang LM, Yan Y, Zou LJ, Jing NH, Xu ZY (2005) Moderate hypothermia prevents neural cell apoptosis following spinal cord ischemia in rabbits. Cell Res 15(5):387–393

    Article  PubMed  CAS  Google Scholar 

  • Wei X, Liu H, Sun X, Fu F, Zhang X, Wang J, An J, Ding H (2005) Hydroxysafflor yellow A protects rat brains against ischemia–reperfusion injury by antioxidant action. Neurosci Lett 386(1):58–62

    Article  PubMed  CAS  Google Scholar 

  • Yang Q, Yang ZF, Liu SB, Zhang XN, Hou Y, Li XQ, Wu YM, Wen AD, Zhao MG (2010) Neuroprotective effects of hydroxysafflor yellow A against excitotoxic neuronal death partially through down-regulation of NR2B-containing NMDA receptors. Neurochem Res 35(9):1353–1360

    Article  PubMed  CAS  Google Scholar 

  • Zhang Y, Bhavnani BR (2005) Glutamate-induced apoptosis in primary cortical neurons is inhibited by equine estrogens via down-regulation of caspase-3 and prevention of mitochondrial cytochrome c release. BMC Neurosci 24:6–13

    Google Scholar 

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Acknowledgment

This study was supported by “the Fundamental Research Funds for the Central Universities” (Program No. xjj20100215).

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Correspondence to Kunzheng Wang.

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Fan, L., Dang, X., Shi, Z. et al. Hydroxysafflor Yellow A Protects PC12 Cells Against the Apoptosis Induced by Oxygen and Glucose Deprivation. Cell Mol Neurobiol 31, 1187–1194 (2011). https://doi.org/10.1007/s10571-011-9720-3

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