Skip to main content

Advertisement

SpringerLink
Log in

Salidroside Inhibits Inflammation Through PI3K/Akt/HIF Signaling After Focal Cerebral Ischemia in Rats

  • ORIGINAL ARTICLE
  • Published:
Inflammation Aims and scope Submit manuscript

Abstract

Salidroside is being investigated for its therapeutic potential in stroke because it is neuroprotective over an extended therapeutic window of time. In the present study, we investigated the mechanisms underlying the anti-inflammatory effects of salidroside (50 mg/kg intraperitoneally) in rats, given 1 h after reperfusion of a middle cerebral artery that had been occluded for 2 h. After 24 h, we found that salidroside increased the neuronal nuclear protein NeuN and reduced the marker of microglia and macrophages CD11b in the peri-infarct area of the brain. Salidroside also decreased IL-6, IL-1β, TNF-α, CD14, CD44, and iNOs mRNAs. At the same time, salidroside increased the ratio of phosphorylated protein kinase B (p-Akt) to total Akt. The phosphoinositide 3-kinase (PI3K) inhibitor LY294002 prevented this increase in p-Akt and reversed the inhibitory effects of salidroside on CD11b and inflammatory mediators. Salidroside also elevated the protein levels of hypoxia-inducible factor (HIF) subunits HIF1α, HIF2α, HIF3α, and of erythropoietin (EPO). The stimulatory effects of salidroside on HIFα subunits were blocked by LY294002. Moreover, YC-1, a HIF inhibitor, abolished salidroside-mediated increase of HIF1α and prevented the inhibitory effects of salidroside on CD11b and inflammatory mediators. Taken together, our results provide evidence for the first time that all three HIFα subunits and EPO can be regulated by PI3K/Akt in cerebral tissue, and that salidroside entrains this signaling pathway to induce production of HIFα subunits and EPO, one or more of which mediate the anti-inflammatory effects of salidroside after cerebral IRI.

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
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  1. Jin, R., G. Yang, and G. Li. 2010. Inflammatory mechanisms in ischemic stroke: role of inflammatory cells. Journal of Leukocyte Biology 87: 779–789.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Hallenbeck, J.M. 1997. Cytokines, macrophages, and leukocytes in brain ischemia. Neurology 49: S5–S9.

    Article  CAS  PubMed  Google Scholar 

  3. Han, H.S., Y. Qiao, M. Karabiyikoglu, R.G. Giffard, and M.A. Yenari. 2002. Influence of mild hypothermia on inducible nitric oxide synthase expression and reactive nitrogen production in experimental stroke and inflammation. Journal of Neuroscience 22: 3921–3928.

    CAS  PubMed  Google Scholar 

  4. Wang, X., L. Xu, H. Wang, Y. Zhan, E. Pure, and G.Z. Feuerstein. 2002. CD44 deficiency in mice protects brain from cerebral ischemia injury. Journal of Neurochemistry 83: 1172–1179.

    Article  CAS  PubMed  Google Scholar 

  5. Muresanu, D.F., A. Buzoianu, S.I. Florian, and T. von Wild. 2012. Towards a roadmap in brain protection and recovery. Journal of Cellular and Molecular Medicine 16: 2861–2871.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Zhang, W. and D. Stanimirovic. 2002. Current and future therapeutic strategies to target inflammation in stroke. Current Drug Targets. Inflammation and Allergy 1: 151–166.

  7. Greer, S.N., J.L. Metcalf, Y. Wang, and M. Ohh. 2012. The updated biology of hypoxia-inducible factor. EMBO Journal 31: 2448–2460.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Weidemann, A., and R.S. Johnson. 2008. Biology of HIF-1alpha. Cell Death and Differentiation 15: 621–627.

    Article  CAS  PubMed  Google Scholar 

  9. Maxwell, P.H., M.S. Wiesener, G.W. Chang, S.C. Clifford, E.C. Vaux, M.E. Cockman, C.C. Wykoff, C.W. Pugh, E.R. Maher, and P.J. Ratcliffe. 1999. The tumour suppressor protein VHL targets hypoxia-inducible factors for oxygen-dependent proteolysis. Nature 399: 271–275.

    Article  CAS  PubMed  Google Scholar 

  10. Joshi, S., A.R. Singh, M. Zulcic, and D.L. Durden. 2014. A macrophage-dominant PI3K isoform controls hypoxia-induced HIF1 and HIF2 stability and tumor growth, angiogenesis, and metastasis. Molecular Cancer Research 12: 1520–1531.

    Article  CAS  PubMed  Google Scholar 

  11. Zhou, J., T. Schmid, R. Frank, and B. Brune. 2004. PI3K/Akt is required for heat shock proteins to protect hypoxia-inducible factor 1alpha from pVHL-independent degradation. Journal of Biological Chemistry 279: 13506–13513.

    Article  CAS  PubMed  Google Scholar 

  12. Treins, C., S. Giorgetti-Peraldi, J. Murdaca, G.L. Semenza, and E. Van Obberghen. 2002. Insulin stimulates hypoxia-inducible factor 1 through a phosphatidylinositol 3-kinase/target of rapamycin-dependent signaling pathway. Journal of Biological Chemistry 277: 27975–27981.

    Article  CAS  PubMed  Google Scholar 

  13. Ye, Z., Q. Guo, P. Xia, N. Wang, E. Wang, and Y. Yuan. 2012. Sevoflurane postconditioning involves an up-regulation of HIF-1alpha and HO-1 expression via PI3K/Akt pathway in a rat model of focal cerebral ischemia. Brain Research 1463: 63–74.

    Article  CAS  PubMed  Google Scholar 

  14. Eltzschig, H.K., D.L. Bratton, and S.P. Colgan. 2014. Targeting hypoxia signalling for the treatment of ischaemic and inflammatory diseases. Nature Reviews Drug Discovery 13: 852–869.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Baranova, O., L.F. Miranda, P. Pichiule, I. Dragatsis, R.S. Johnson, and J.C. Chavez. 2007. Neuron-specific inactivation of the hypoxia inducible factor 1 increases brain injury in a mouse model of transient focal cerebral ischemia. Journal of Neuroscience 27: 6320–6332.

    Article  CAS  PubMed  Google Scholar 

  16. Shi, H. 2009. Hypoxia inducible factor 1 as a therapeutic target in ischemic stroke. Current Medicinal Chemistry 16: 4593–4600.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Wang, C., Z. Wang, X. Zhang, X. Zhang, L. Dong, Y. Xing, Y. Li, Z. Liu, L. Chen, H. Qiao, L. Wang, and C. Zhu. 2012. Protection by silibinin against experimental ischemic stroke: up-regulated pAkt, pmTOR, HIF-1α and Bcl-2, down-regulated Bax, NF-κB expression. Neuroscience Letters 529: 45–50.

    Article  CAS  PubMed  Google Scholar 

  18. Lai, W.F., Z.W. Zheng, X.Q. Zhang, Y.C. Wei, K.D. Chu, J. Brown, G.Z. Hong, and L.D. Chen. 2015. Salidroside-mediated neuroprotection is associated with induction of early growth response genes (Egrs) across a wide therapeutic window. Neurotoxicity Research 28: 108–121.

    Article  CAS  PubMed  Google Scholar 

  19. Shi, T.Y., S.F. Feng, J.H. Xing, Y.M. Wu, X.Q. Li, N. Zhang, Z. Tian, S.B. Liu, and M.G. Zhao. 2012. Neuroprotective effects of salidroside and its analogue tyrosol galactoside against focal cerebral ischemia in vivo and H2O2-induced neurotoxicity in vitro. Neurotoxicity Research 21: 358–367.

    Article  CAS  PubMed  Google Scholar 

  20. Zhu, L., T. Wei, X. Chang, H. He, J. Gao, Z. Wen, and T. Yan. 2015. Effects of salidroside on myocardial injury in vivo in vitro via regulation of Nox/NF-kappaB/AP1 pathway. Inflammation 38: 1589–1598.

    Article  CAS  PubMed  Google Scholar 

  21. Hu, H., Z. Li, X. Zhu, R. Lin, and L. Chen. 2014. Salidroside reduces cell mobility via NF-κB and MAPK signaling in LPS-induced BV2 microglial cells. Evidence-based Complementary and Alternative Medicine 2014: 383821.

    PubMed  PubMed Central  Google Scholar 

  22. Gao, J., R. Zhou, X. You, F. Luo, H. He, X. Chang, L. Zhu, X. Ding, and T. Yan. 2016. Salidroside suppresses inflammation in a D-galactose-induced rat model of Alzheimer’s disease via SIRT1/NF-kappaB pathway. Metabolic Brain Disease 31: 771–778.

    Article  CAS  PubMed  Google Scholar 

  23. Zheng, K.Y., Z.X. Zhang, A.J. Guo, C.W. Bi, K.Y. Zhu, S.L. Xu, J.Y. Zhan, D.T. Lau, T.T. Dong, R.C. Choi, and K.W. Tsim. 2012. Salidroside stimulates the accumulation of HIF-1alpha protein resulted in the induction of EPO expression: a signaling via blocking the degradation pathway in kidney and liver cells. European Journal of Pharmacology 679: 34–39.

    Article  CAS  PubMed  Google Scholar 

  24. Longa, E.Z., P.R. Weinstein, S. Carlson, and R. Cummins. 1989. Reversible middle cerebral artery occlusion without craniectomy in rats. Stroke 20: 84–91.

    Article  CAS  PubMed  Google Scholar 

  25. Nakayama, H., M.D. Ginsberg, and W.D. Dietrich. 1988. (S)-emopamil, a novel calcium channel blocker and serotonin S2 antagonist, markedly reduces infarct size following middle cerebral artery occlusion in the rat. Neurology 38: 1667–1673.

    Article  CAS  PubMed  Google Scholar 

  26. Belayev, L., O.F. Alonso, R. Busto, W. Zhao, and M.D. Ginsberg. 1996. Middle cerebral artery occlusion in the rat by intraluminal suture. Neurological and pathological evaluation of an improved model. Stroke 27 (1616–1622): 1623.

    Google Scholar 

  27. Zhao, H., T. Shimohata, J.Q. Wang, G. Sun, D.W. Schaal, R.M. Sapolsky, and G.K. Steinberg. 2005. Akt contributes to neuroprotection by hypothermia against cerebral ischemia in rats. Journal of Neuroscience 25: 9794–9806.

    Article  CAS  PubMed  Google Scholar 

  28. Zhang, Z., J. Yan, S. Taheri, K.J. Liu, and H. Shi. 2014. Hypoxia-inducible factor 1 contributes to N-acetylcysteine’s protection in stroke. Free Radical Biology and Medicine 68: 8–21.

    Article  CAS  PubMed  Google Scholar 

  29. Sun, M., B. Deng, X. Zhao, C. Gao, L. Yang, H. Zhao, D. Yu, F. Zhang, L. Xu, L. Chen, and X. Sun. 2015. Isoflurane preconditioning provides neuroprotection against stroke by regulating the expression of the TLR4 signalling pathway to alleviate microglial activation. Scientific Reports 5: 11445.

    Article  PubMed  PubMed Central  Google Scholar 

  30. Perego, C., S. Fumagalli, and M.G. De Simoni. 2011. Temporal pattern of expression and colocalization of microglia/macrophage phenotype markers following brain ischemic injury in mice. Journal of Neuroinflammation 8: 174.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Guha, M., and N. Mackman. 2002. The phosphatidylinositol 3-kinase-Akt pathway limits lipopolysaccharide activation of signaling pathways and expression of inflammatory mediators in human monocytic cells. Journal of Biological Chemistry 277: 32124–32132.

    Article  CAS  PubMed  Google Scholar 

  32. Jin, R., Z. Song, S. Yu, A. Piazza, A. Nanda, J.M. Penninger, D.N. Granger, and G. Li. 2011. Phosphatidylinositol-3-kinase gamma plays a central role in blood-brain barrier dysfunction in acute experimental stroke. Stroke 42: 2033–2044.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Lai, W.F., X. Tian, Q. Xiang, K.D. Chu, Y. Wei, J.T. Deng, S.P. Zhang, J. Brown, and G.Z. Hong. 2015. 11β-HSD1 modulates LPS-induced innate immune responses in adipocytes by altering expression of PTEN. Molecular Endocrinology 29: 558–570.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Li, L., D.W. McBride, D. Doycheva, B.J. Dixon, P.R. Krafft, J.H. Zhang, and J.P. Tang. 2015. G-CSF attenuates neuroinflammation and stabilizes the blood–brain barrier via the PI3K/Akt/GSK-3β signaling pathway following neonatal hypoxia-ischemia in rats. Experimental Neurology 272: 135–144.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Chun, Y.S., E.J. Yeo, E. Choi, C.M. Teng, J.M. Bae, M.S. Kim, and J.W. Park. 2001. Inhibitory effect of YC-1 on the hypoxic induction of erythropoietin and vascular endothelial growth factor in Hep3B cells. Biochemical Pharmacology 61: 947–954.

    Article  CAS  PubMed  Google Scholar 

  36. Li, S.H., D.H. Shin, Y.S. Chun, M.K. Lee, M.S. Kim, and J.W. Park. 2008. A novel mode of action of YC-1 in HIF inhibition: stimulation of FIH-dependent p300 dissociation from HIF-1. Molecular Cancer Therapeutics 7: 3729–3738.

    Article  CAS  PubMed  Google Scholar 

  37. Zhang, L., W. Ding, H. Sun, Q. Zhou, J. Huang, X. Li, Y. Xie, and J. Chen. 2012. Salidroside protects PC12 cells from MPP(+)-induced apoptosis via activation of the PI3K/Akt pathway. Food and Chemical Toxicology 50: 2591–2597.

    Article  CAS  PubMed  Google Scholar 

  38. Zhang, B., Y. Wang, H. Li, R. Xiong, Z. Zhao, X. Chu, Q. Li, S. Sun, and S. Chen. 2016. Neuroprotective effects of salidroside through PI3K/Akt pathway activation in Alzheimer’s disease models. Drug Design, Development and Therapy 10: 1335–1343.

    PubMed  PubMed Central  Google Scholar 

  39. Chen, S.F., H.J. Tsai, T.H. Hung, C.C. Chen, C.Y. Lee, C.H. Wu, P.Y. Wang, and N.C. Liao. 2012. Salidroside improves behavioral and histological outcomes and reduces apoptosis via PI3K/Akt signaling after experimental traumatic brain injury. PloS One 7: e45763.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Zhang, W., H. He, H. Song, J. Zhao, T. Li, L. Wu, X. Zhang, and J. Chen. 2016. Neuroprotective effects of salidroside in the MPTP mouse model of Parkinson’s disease: involvement of the PI3K/Akt/GSK3beta pathway. Parkinson's Disease 2016: 9450137.

    Article  PubMed  PubMed Central  Google Scholar 

  41. Guan, S., H. Feng, B. Song, W. Guo, Y. Xiong, G. Huang, W. Zhong, M. Huo, N. Chen, J. Lu, and X. Deng. 2011. Salidroside attenuates LPS-induced pro-inflammatory cytokine responses and improves survival in murine endotoxemia. International Immunopharmacology 11: 2194–2199.

    Article  CAS  PubMed  Google Scholar 

  42. Zhu, L., T. Wei, J. Gao, X. Chang, H. He, M. Miao, and T. Yan. 2015. Salidroside attenuates lipopolysaccharide (LPS) induced serum cytokines and depressive-like behavior in mice. Neuroscience Letters 606: 1–6.

    Article  CAS  PubMed  Google Scholar 

  43. Wu, D., P. Yuan, C. Ke, H. Xiong, J. Chen, J. Guo, M. Lu, Y. Ding, X. Fan, Q. Duan, F. Shi, and F. Zhu. 2016. Salidroside suppresses solar ultraviolet-induced skin inflammation by targeting cyclooxygenase-2. Oncotarget 7: 25971–25982.

    PubMed  PubMed Central  Google Scholar 

  44. Sun, P., S.Z. Song, S. Jiang, X. Li, Y.L. Yao, Y.L. Wu, L.H. Lian, J.X. Nan. 2016. Salidroside regulates inflammatory response in Raw 264.7 macrophages via TLR4/TAK1 and ameliorates inflammation in alcohol binge drinking-induced liver injury. Molecules 21:E 1490.

  45. Sun, H.L., Y.N. Liu, Y.T. Huang, S.L. Pan, D.Y. Huang, J.H. Guh, F.Y. Lee, S.C. Kuo, and C.M. Teng. 2007. YC-1 inhibits HIF-1 expression in prostate cancer cells: contribution of Akt/NF-kappaB signaling to HIF-1alpha accumulation during hypoxia. Oncogene 26: 3941–3951.

    Article  CAS  PubMed  Google Scholar 

  46. Ivan, M., K. Kondo, H. Yang, W. Kim, J. Valiando, M. Ohh, A. Salic, J.M. Asara, W.S. Lane, and W.J. Kaelin. 2001. HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. Science 292: 464–468.

    Article  CAS  PubMed  Google Scholar 

  47. Heidbreder, M., F. Frohlich, O. Johren, A. Dendorfer, F. Qadri, and P. Dominiak. 2003. Hypoxia rapidly activates HIF-3alpha mRNA expression. FASEB Journal 17: 1541–1543.

    CAS  PubMed  Google Scholar 

  48. Zhang, P., Q. Yao, L. Lu, Y. Li, P.J. Chen, and C. Duan. 2014. Hypoxia-inducible factor 3 is an oxygen-dependent transcription activator and regulates a distinct transcriptional response to hypoxia. Cell Reports 6: 1110–1121.

    Article  CAS  PubMed  Google Scholar 

  49. Sarada, S., M. Titto, P. Himadri, S. Saumya, and V. Vijayalakshmi. 2015. Curcumin prophylaxis mitigates the incidence of hypobaric hypoxia-induced altered ion channels expression and impaired tight junction proteins integrity in rat brain. Journal of Neuroinflammation 12: 113.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Koh, H.S., C.Y. Chang, S. Jeon, H.J. Yoon, Y. Ahn, H. Kim, I. Kim, S.H. Jeon, R.S. Johnson, and E.J. Park. 2015. The HIF-1/glial TIM-3 axis controls inflammation-associated brain damage under hypoxia. Nature Communications 6: 6340.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Villa, P., P. Bigini, T. Mennini, D. Agnello, T. Laragione, A. Cagnotto, B. Viviani, M. Marinovich, A. Cerami, T.R. Coleman, M. Brines, and P. Ghezzi. 2003. Erythropoietin selectively attenuates cytokine production and inflammation in cerebral ischemia by targeting neuronal apoptosis. Journal of Experimental Medicine 198: 971–975.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Marti, H.H., R.H. Wenger, L.A. Rivas, U. Straumann, M. Digicaylioglu, V. Henn, Y. Yonekawa, C. Bauer, and M. Gassmann. 1996. Erythropoietin gene expression in human, monkey and murine brain. European Journal of Neuroscience 8: 666–676.

    Article  CAS  PubMed  Google Scholar 

  53. Masuda, S., M. Okano, K. Yamagishi, M. Nagao, M. Ueda, R. Sasaki. 1994. A novel site of erythropoietin production. Oxygen-dependent production in cultured rat astrocytes. Journal of Biological Chemistry 269: 19488–19493.

  54. Rabie, T., and H.H. Marti. 2008. Brain protection by erythropoietin: a manifold task. Physiology 23: 263–274.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by the National Natural Science Foundation of China (Grant No. 81473382), the Department of Technology and Science and the Department of Health of Fujian Provincial Government (Grant Nos. 2014Y4004 and WZZY201305), the Collaborative Innovation Center for Rehabilitation Technology of Fujian University of TCM, and the TCM Rehabilitation Research Center of SATCM. The authors would like to acknowledge the staff of the Animal Centre of the Fujian University of TCM for their excellent technical support.

Author information

Authors and Affiliations

  1. Center of Biomedical Research & Development, Fujian University of Traditional Chinese Medicine, No. 1 Huatou Road, Minhou Shangjie, Fuzhou, China

    Yicong Wei, Haimian Hong, Xiaoqin Zhang, Wenfang Lai, Yingzheng Wang, Kedan Chu, John Brown, Guizhu Hong & Lidian Chen

Authors
  1. Yicong Wei
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Haimian Hong
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xiaoqin Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Wenfang Lai
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yingzheng Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Kedan Chu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. John Brown
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Guizhu Hong
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Lidian Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lidian Chen.

Ethics declarations

Conflict of Interest

The authors declare that they have no competing interests.

Additional information

Yicong Wei and Haimian Hong contributed equally to this work

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Wei, Y., Hong, H., Zhang, X. et al. Salidroside Inhibits Inflammation Through PI3K/Akt/HIF Signaling After Focal Cerebral Ischemia in Rats. Inflammation 40, 1297–1309 (2017). https://doi.org/10.1007/s10753-017-0573-x

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10753-017-0573-x

KEY WORDS

Search

Navigation