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Cellular and Molecular Neurobiology

, Volume 36, Issue 8, pp 1365–1375 | Cite as

Saikosaponin-D Reduces H2O2-Induced PC12 Cell Apoptosis by Removing ROS and Blocking MAPK-Dependent Oxidative Damage

  • Xuemei Lin
  • Songdi Wu
  • Qing Wang
  • Yaling Shi
  • Guozheng Liu
  • Jin Zhi
  • Fang Wang
Original Research

Abstract

Neuronal oxidative stress (OS) injury has been proven to be associated with many neurodegenerative diseases, and thus, antioxidation treatment is an effective method for treating these diseases. Saikosaponin-D (SSD) is a sapogenin extracted from Bupleurum falcatum and has been shown to have many pharmacological activities. The main purpose of this study was to investigate whether and how SSD protects PC12 cells from H2O2-induced apoptosis. The non-toxic level of SSD significantly mitigated the H2O2-induced decrease in cell viability, reduced the apoptosis rate, improved the nuclear morphology, and reduced caspase-3 activation and poly ADP-ribose polymerase (PARP) cleavage. Additionally, exogenous H2O2-induced apoptosis by damaging the intracellular antioxidation system. SSD significantly slowed the H2O2-induced release of malonic dialdehyde (MDA) and lactate dehydrogenase and increased the activity of superoxide dismutase (SOD) and the total antioxidant capacity, thereby reducing apoptosis. More importantly, SSD effectively blocked H2O2-induced phosphorylation of extracellular-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (p38MAPK), and specific inhibitors of ERK, JNK, and p38-reduced OS injury and apoptosis, suggesting that SSD reduces OS injury and apoptosis via MAPK signalling pathways. Finally, we confirmed that SSD significantly reduced H2O2-induced reactive oxygen species (ROS) accumulation, and the ROS inhibitor blocked the apoptosis caused by MAPK activation and cellular oxidative damage. In short, our study confirmed that SSD reduces H2O2-induced PC12 cell apoptosis by removing ROS and blocking MAPK-dependent oxidative damage.

Keywords

Saikosaponin-D Oxidative stress Cell apoptosis Reactive oxygen species PC12 cell 

References

  1. Aminzadeh A, Dehpour AR, Safa M, Mirzamohammadi S, Sharifi AM (2014) Investigating the protective effect of lithium against high glucose-induced neurotoxicity in PC12 cells: involvements of ROS, JNK and P38 MAPKs, and apoptotic mitochondria pathway. Cell Mol Neurobiol 34(8):1143–1150. doi: 10.1007/s10571-014-0089-y CrossRefPubMedGoogle Scholar
  2. Ankarcrona M, Dypbukt JM, Bonfoco E, Zhivotovsky B, Orrenius S, Lipton SA, Nicotera P (1995) Glutamate-induced neuronal death: a succession of necrosis or apoptosis depending on mitochondrial function. Neuron 15(4):961–973CrossRefPubMedGoogle Scholar
  3. Barnham KJ, Masters CL, Bush AI (2004) Neurodegenerative diseases and oxidative stress. Nat Rev Drug Discov 3(3):205–214. doi: 10.1038/nrd1330 CrossRefPubMedGoogle Scholar
  4. Chen XH, Zhou X, Yang XY, Zhou ZB, Lu DH, Tang Y et al (2015) Propofol protects against HO—induced oxidative injury in differentiated PC12 Cells via inhibition of Ca-dependent NADPH oxidase. Cell Mol Neurobiol. doi: 10.1007/s10571-015-0235-1 Google Scholar
  5. Cui C, Cui N, Wang P, Song S, Liang H, Ji A (2015) Sulfated polysaccharide isolated from the sea cucumber Stichopus japonicus against PC12 hypoxia/reoxygenation injury by inhibition of the MAPK signaling pathway. Cell Mol Neurobiol. doi: 10.1007/s10571-015-0202-x Google Scholar
  6. Finkel T, Holbrook NJ (2000) Oxidants, oxidative stress and the biology of ageing. Nature 408(6809):239–247. doi: 10.1038/35041687 CrossRefPubMedGoogle Scholar
  7. Fransen M, Nordgren M, Wang B, Apanasets O (2012) Role of peroxisomes in ROS/RNS-metabolism: implications for human disease. Biochim Biophys Acta 1822(9):1363–1373. doi: 10.1016/j.bbadis.2011.12.001 CrossRefPubMedGoogle Scholar
  8. Galluzzi L, Kepp O, Kroemer G (2012) Caspase-3 and prostaglandins signal for tumor regrowth in cancer therapy. Oncogene 31(23):2805–2808. doi: 10.1038/onc.2011.459 CrossRefPubMedGoogle Scholar
  9. Gao Y, Dong C, Yin J, Shen J, Tian J, Li C (2012) Neuroprotective effect of fucoidan on H2O2-induced apoptosis in PC12 cells via activation of PI3K/Akt pathway. Cell Mol Neurobiol 32(4):523–529. doi: 10.1007/s10571-011-9792-0 CrossRefPubMedGoogle Scholar
  10. Kim H, Youn K, Ahn MR, Kim OY, Jeong WS, Ho CT, Jun M (2015) Neuroprotective effect of loganin against Abeta25-35-induced injury via the NF-kappaB-dependent signaling pathway in PC12 cells. Food Funct 6(4):1108–1116. doi: 10.1039/c5fo00055f CrossRefPubMedGoogle Scholar
  11. Li ZY, Guo Z, Liu YM, Liu XM, Chang Q, Liao YH, Pan RL (2013) Neuroprotective effects of total saikosaponins of Bupleurum yinchowense on corticosterone-induced apoptosis in PC12 cells. J Ethnopharmacol 148(3):794–803. doi: 10.1016/j.jep.2013.04.057 CrossRefPubMedGoogle Scholar
  12. Li ZY, Jiang YM, Liu YM, Guo Z, Shen SN, Liu XM, Pan RL (2014) Saikosaponin D acts against corticosterone-induced apoptosis via regulation of mitochondrial GR translocation and a GR-dependent pathway. Prog Neuropsychopharmacol Biol Psychiatry 53:80–89. doi: 10.1016/j.pnpbp.2014.02.010 CrossRefPubMedGoogle Scholar
  13. Lin TY, Chiou CY, Chiou SJ (2013) Putative genes involved in saikosaponin biosynthesis in bupleurum species. Int J Mol Sci 14(6):12806–12826. doi: 10.3390/ijms140612806 CrossRefPubMedPubMedCentralGoogle Scholar
  14. Lu CN, Yuan ZG, Zhang XL, Yan R, Zhao YQ, Liao M, Chen JX (2012) Saikosaponin a and its epimer saikosaponin d exhibit anti-inflammatory activity by suppressing activation of NF-kappaB signaling pathway. Int Immunopharmacol 14(1):121–126. doi: 10.1016/j.intimp.2012.06.010 CrossRefPubMedGoogle Scholar
  15. Luo Y, Liu X, Zheng Q, Wan X, Ouyang S, Yin Y et al (2012) Hydrogen sulfide prevents hypoxia-induced apoptosis via inhibition of an H2O2-activated calcium signaling pathway in mouse hippocampal neurons. Biochem Biophys Res Commun 425(2):473–477. doi: 10.1016/j.bbrc.2012.07.131 CrossRefPubMedGoogle Scholar
  16. Ma S, Liu X, Xun Q, Zhang X (2014) Neuroprotective effect of Ginkgolide K against H2O2-induced PC12 cell cytotoxicity by ameliorating mitochondrial dysfunction and oxidative stress. Biol Pharm Bull 37(2):217–225CrossRefPubMedGoogle Scholar
  17. Ma X, Dang C, Kang H, Dai Z, Lin S, Guan H et al (2015) Saikosaponin-d reduces cisplatin-induced nephrotoxicity by repressing ROS-mediated activation of MAPK and NF-kappaB signalling pathways. Int Immunopharmacol 28(1):399–408. doi: 10.1016/j.intimp.2015.06.020 CrossRefPubMedGoogle Scholar
  18. Maruyama W, Shaomoto-Nagai M, Kato Y, Hisaka S, Osawa T, Naoi M (2014) Role of lipid peroxide in the neurodegenerative disorders. Subcell Biochem 77:127–136. doi: 10.1007/978-94-007-7920-4_11 CrossRefPubMedGoogle Scholar
  19. Tian X, Sui S, Huang J, Bai JP, Ren TS, Zhao QC (2014) Neuroprotective effects of Arctium lappa L. roots against glutamate-induced oxidative stress by inhibiting phosphorylation of p38, JNK and ERK 1/2 MAPKs in PC12 cells. Environ Toxicol Pharmacol 38(1):189–198. doi: 10.1016/j.etap.2014.05.017 CrossRefPubMedGoogle Scholar
  20. Westerblad H, Allen DG (2011) Emerging roles of ROS/RNS in muscle function and fatigue. Antioxid Redox Signal 15(9):2487–2499. doi: 10.1089/ars.2011.3909 CrossRefPubMedGoogle Scholar
  21. Wong VK, Zhang MM, ZhouH Lam KY, Chan PL, Law CK, Liu L (2013) Saikosaponin-d enhances the anticancer potency of TNF-alpha via overcoming its undesirable response of activating NF-Kappa B signalling in cancer cells. Evid Based Complement Altern Med 2013:745295. doi: 10.1155/2013/745295 Google Scholar
  22. Wu SJ, Tam KW, Tsai YH, Chang CC, Chao JC (2010) Curcumin and saikosaponin a inhibit chemical-induced liver inflammation and fibrosis in rats. Am J Chin Med 38(1):99–111. doi: 10.1142/S0192415X10007695 CrossRefPubMedGoogle Scholar
  23. Zawia NH, Lahiri DK, Cardozo-Pelaez F (2009) Epigenetics, oxidative stress, and Alzheimer disease. Free Radic Biol Med 46(9):1241–1249. doi: 10.1016/j.freeradbiomed.2009.02.006 CrossRefPubMedPubMedCentralGoogle Scholar
  24. Zhao ZY, Luan P, Huang SX, Xiao SH, Zhao J, Zhang B, Liu J (2013) Edaravone protects HT22 neurons from H2O2-induced apoptosis by inhibiting the MAPK signaling pathway. CNS Neurosci Ther 19(3):163–169. doi: 10.1111/cns.12044 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Xuemei Lin
    • 1
  • Songdi Wu
    • 1
  • Qing Wang
    • 1
  • Yaling Shi
    • 1
  • Guozheng Liu
    • 1
  • Jin Zhi
    • 1
  • Fang Wang
    • 1
  1. 1.Department of NeurologyThe First Hospital of Xi’anXi’anChina

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