Skip to main content
SpringerLink
Log in

Hydroxysafflor Yellow A Protects Against Cerebral Ischemia–Reperfusion Injury by Anti-apoptotic Effect Through PI3K/Akt/GSK3β Pathway in Rat

  • Original Paper
  • Published:
Neurochemical Research Aims and scope Submit manuscript

Abstract

Hydroxysafflor yellow A (HSYA) is the major active chemical component of the flower of the safflower plant, Carthamus tinctorius L. Previously, its neuroprotection against cerebral ischemia–reperfusion (I/R) injury was reported by anti-oxidant action and suppression of thrombin generation. Here, we investigate the role of HSYA in cerebral I/R-mediated apoptosis and possible signaling pathways. Male Wistar rats were subjected to transient middle cerebral artery occlusion for 2 h, followed by 24 h reperfusion. HSYA was administered via tail-vein injection just 15 min after occlusion. The number of apoptotic cells was measured by TUNEL assay, apoptosis-related proteins Bcl-2, Bax and the phosphorylation levels of Akt and GSK3β in ischemic penumbra were assayed by western blot. The results showed that administration of HSYA at the doses of 4 and 8 mg/kg significantly inhibited the apoptosis by decreasing the number of apoptotic cells and increasing the Bcl-2/Bax ratio in rats subjected to I/R injury. Simultaneously, HSYA treatment markedly increased the phosphorylations of Akt and GSK3β. Blockade of PI3K activity by wortmannin dramatically abolished its anti-apoptotic effect and lowered both Akt and GSK3β phosphorylation levels. Taken together, these results suggest that HSYA protects against cerebral I/R injury partly by reducing apoptosis via PI3K/Akt/GSK3β signaling pathway.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Dirnagl U, Iadecola C, Moskowitz MA (1999) Pathobiology of ischaemic stroke: an integrated view. Trends Neurosci 22(9):391–397

    Article  PubMed  CAS  Google Scholar 

  2. Broughton BR, Reutens DC, Sobey CG (2009) Apoptotic mechanisms after cerebral ischemia. Stroke 40(5):e331–e339. doi:10.1161/STROKEAHA.108.531632

    Article  PubMed  Google Scholar 

  3. Sun B, Chen L, Wei X, Xiang Y, Liu X, Zhang X (2011) The Akt/GSK-3beta pathway mediates flurbiprofen-induced neuroprotection against focal cerebral ischemia/reperfusion injury in rats. Biochem Biophys Res Commun 409(4):808–813. doi:10.1016/j.bbrc.2011.05.095

    Article  PubMed  CAS  Google Scholar 

  4. Liu H, Liu X, Wei X, Chen L, Xiang Y, Yi F, Zhang X (2012) Losartan, an angiotensin II type 1 receptor blocker, ameliorates cerebral ischemia–reperfusion injury via PI3K/Akt-mediated eNOS phosphorylation. Brain Res Bull 89(1–2):65–70. doi:10.1016/j.brainresbull.2012.06.010

    Article  PubMed  CAS  Google Scholar 

  5. Zhang J, Deng Z, Liao J, Song C, Liang C, Xue H, Wang L, Zhang K, Yan G (2013) Leptin attenuates cerebral ischemia injury through the promotion of energy metabolism via the PI3K/Akt pathway. J Cereb Blood Flow Metab 33(4):567–574. doi:10.1038/jcbfm.2012.202

    Article  PubMed  CAS  Google Scholar 

  6. Noshita N, Lewen A, Sugawara T, Chan PH (2001) Evidence of phosphorylation of Akt and neuronal survival after transient focal cerebral ischemia in mice. J Cereb Blood Flow Metab 21(12):1442–1450. doi:10.1097/00004647-200112000-00009

    Article  PubMed  CAS  Google Scholar 

  7. Cantley LC (2002) The phosphoinositide 3-kinase pathway. Science 296(5573):1655–1657. doi:10.1126/science.296.5573.1655

    Article  PubMed  CAS  Google Scholar 

  8. Martin M, Rehani K, Jope RS, Michalek SM (2005) Toll-like receptor-mediated cytokine production is differentially regulated by glycogen synthase kinase 3. Nat Immunol 6(8):777–784. doi:10.1038/ni1221

    Article  PubMed  CAS  Google Scholar 

  9. Jope RS, Johnson GV (2004) The glamour and gloom of glycogen synthase kinase-3. Trends Biochem Sci 29(2):95–102. doi:10.1016/j.tibs.2003.12.004

    Article  PubMed  CAS  Google Scholar 

  10. Beurel E, Jope RS (2006) The paradoxical pro- and anti-apoptotic actions of GSK3 in the intrinsic and extrinsic apoptosis signaling pathways. Prog Neurobiol 79(4):173–189. doi:10.1016/j.pneurobio.2006.07.006

    Article  PubMed  CAS  Google Scholar 

  11. Zhao H, Sapolsky RM, Steinberg GK (2006) Phosphoinositide-3-kinase/akt survival signal pathways are implicated in neuronal survival after stroke. Mol Neurobiol 34(3):249–270. doi:10.1385/MN:34:3:249

    Article  PubMed  CAS  Google Scholar 

  12. Zhu H, Wang Z, Ma C, Tian J, Fu F, Li C, Guo D, Roeder E, Liu K (2003) Neuroprotective effects of hydroxysafflor yellow A: in vivo and in vitro studies. Planta Med 69(5):429–433. doi:10.1055/s-2003-39714

    Article  PubMed  CAS  Google Scholar 

  13. 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. doi:10.1016/j.neulet.2005.05.069

    Article  PubMed  CAS  Google Scholar 

  14. Sun X, Wei X, Qu S, Zhao Y, Zhang X (2010) Hydroxysafflor Yellow A suppresses thrombin generation and inflammatory responses following focal cerebral ischemia–reperfusion in rats. Bioorg Med Chem Lett 20(14):4120–4124. doi:10.1016/j.bmcl.2010.05.076

    Article  PubMed  CAS  Google Scholar 

  15. 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. doi:10.1186/1471-2202-11-98

    Article  PubMed  Google Scholar 

  16. 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. doi:10.1007/s11064-010-0191-6

    Article  PubMed  CAS  Google Scholar 

  17. Chen L, Zhang Y, Sun X, Li H, LeSage G, Javer A, Zhang X, Wei X, Jiang Y, Yin D (2009) Synthetic resveratrol aliphatic acid inhibits TLR2-mediated apoptosis and an involvement of Akt/GSK3beta pathway. Bioorg Med Chem 17(13):4378–4382. doi:10.1016/j.bmc.2009.05.029

    Article  PubMed  CAS  Google Scholar 

  18. Chen L, Shi W, Li H, Sun X, Fan X, Lesage G, Li Y, Zhang Y, Zhang X, Yin D (2010) Critical role of toll-like receptor 9 in morphine and Mycobacterium tuberculosis-Induced apoptosis in mice. PLoS ONE 5(2):e9205. doi:10.1371/journal.pone.0009205

    Article  PubMed  Google Scholar 

  19. Benchoua A, Guegan C, Couriaud C, Hosseini H, Sampaio N, Morin D, Onteniente B (2001) Specific caspase pathways are activated in the two stages of cerebral infarction. J Neurosci 21(18):7127–7134

    PubMed  CAS  Google Scholar 

  20. Ferrer I, Friguls B, Dalfo E, Justicia C, Planas AM (2003) Caspase-dependent and caspase-independent signalling of apoptosis in the penumbra following middle cerebral artery occlusion in the adult rat. Neuropathol Appl Neurobiol 29(5):472–481

    Article  PubMed  CAS  Google Scholar 

  21. Sugawara T, Chan PH (2003) Reactive oxygen radicals and pathogenesis of neuronal death after cerebral ischemia. Antioxid Redox Signal 5(5):597–607. doi:10.1089/152308603770310266

    Article  PubMed  CAS  Google Scholar 

  22. 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. doi:10.1016/j.vph.2008.11.009

    Article  PubMed  CAS  Google Scholar 

  23. 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. doi:10.1097/FJC.0b013e318181fb02

    Article  PubMed  CAS  Google Scholar 

  24. Murphy KM, Streips UN, Lock RB (1999) Bax membrane insertion during Fas(CD95)-induced apoptosis precedes cytochrome c release and is inhibited by Bcl-2. Oncogene 18(44):5991–5999. doi:10.1038/sj.onc.1203001

    Article  PubMed  CAS  Google Scholar 

  25. Li F, Omori N, Jin G, Wang SJ, Sato K, Nagano I, Shoji M, Abe K (2003) Cooperative expression of survival p-ERK and p-Akt signals in rat brain neurons after transient MCAO. Brain Res 962(1–2):21–26

    PubMed  CAS  Google Scholar 

  26. Friguls B, Justicia C, Pallas M, Planas AM (2001) Focal cerebral ischemia causes two temporal waves of Akt activation. NeuroReport 12(15):3381–3384

    Article  PubMed  CAS  Google Scholar 

  27. Liu SX, Zhang Y, Wang YF, Li XC, Xiang MX, Bian C, Chen P (2012) Upregulation of heme oxygenase-1 expression by hydroxysafflor yellow A conferring protection from anoxia/reoxygenation-induced apoptosis in H9c2 cardiomyocytes. Int J Cardiol 160(2):95–101. doi:10.1016/j.ijcard.2011.03.033

    Article  PubMed  Google Scholar 

  28. Yu SM, Kim SJ (2013) Thymoquinone-induced reactive oxygen species causes apoptosis of chondrocytes via PI3K/Akt and p38kinase pathway. Exp Biol Med (Maywood). doi:10.1177/1535370213492685

    Google Scholar 

  29. Kwon SH, Hong SI, Kim JA, Jung YH, Kim SY, Kim HC, Lee SY, Jang CG (2011) The neuroprotective effects of Lonicera japonica THUNB. against hydrogen peroxide-induced apoptosis via phosphorylation of MAPKs and PI3K/Akt in SH-SY5Y cells. Food Chem Toxicol 49(4):1011–1019. doi:10.1016/j.fct.2011.01.008

    Article  PubMed  CAS  Google Scholar 

  30. Woodgett JR (2005) Recent advances in the protein kinase B signaling pathway. Curr Opin Cell Biol 17(2):150–157. doi:10.1016/j.ceb.2005.02.010

    Article  PubMed  CAS  Google Scholar 

  31. Leroy K, Brion JP (1999) Developmental expression and localization of glycogen synthase kinase-3beta in rat brain. J Chem Neuroanat 16(4):279–293

    Article  PubMed  CAS  Google Scholar 

  32. Pap M, Cooper GM (1998) Role of glycogen synthase kinase-3 in the phosphatidylinositol 3-Kinase/Akt cell survival pathway. J Biol Chem 273(32):19929–19932

    Article  PubMed  CAS  Google Scholar 

  33. Linseman DA, Butts BD, Precht TA, Phelps RA, Le SS, Laessig TA, Bouchard RJ, Florez-McClure ML, Heidenreich KA (2004) Glycogen synthase kinase-3beta phosphorylates Bax and promotes its mitochondrial localization during neuronal apoptosis. J Neurosci 24(44):9993–10002. doi:10.1523/JNEUROSCI.2057-04.2004

    Article  PubMed  CAS  Google Scholar 

  34. Kaga S, Zhan L, Altaf E, Maulik N (2006) Glycogen synthase kinase-3beta/beta-catenin promotes angiogenic and anti-apoptotic signaling through the induction of VEGF, Bcl-2 and surviving expression in rat ischemic preconditioned myocardium. J Mol Cell Cardiol 40(1):138–147. doi:10.1016/j.yjmcc.2005.09.009

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was financially supported by National Natural Science Foundation of China (No. 81100874, No. 81200904), Young Excellent Scientists of Shandong Province (No. BS2010YY037, 2011BSE27078) and Natural Science Foundation of Shandong Province (No. ZR2012HM032, ZR2010HM082).

Author information

Authors and Affiliations

  1. Department of Pharmacology, Shandong University School of Medicine, 44#, Wenhua Xi Road, Jinan, 250012, Shandong, People’s Republic of China

    Lin Chen, Yanxiao Xiang, Lingjun Kong, Xiumei Zhang, Xinbing Wei & Huiqing Liu

  2. Department of Anesthesiology, Qilu Hospital, Shandong University, 107#, Wenhua Xi Road, Jinan, 250012, Shandong, People’s Republic of China

    Baozhu Sun

Authors
  1. Lin Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yanxiao Xiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Lingjun Kong
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Xiumei Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Baozhu Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Xinbing Wei
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Huiqing Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Xinbing Wei or Huiqing Liu.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chen, L., Xiang, Y., Kong, L. et al. Hydroxysafflor Yellow A Protects Against Cerebral Ischemia–Reperfusion Injury by Anti-apoptotic Effect Through PI3K/Akt/GSK3β Pathway in Rat. Neurochem Res 38, 2268–2275 (2013). https://doi.org/10.1007/s11064-013-1135-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11064-013-1135-8

Keywords

Search

Navigation