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Tetramethylpyrazine Protects Against Early Brain Injury and Inhibits the PERK/Akt Pathway in a Rat Model of Subarachnoid Hemorrhage

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Abstract

Neuronal apoptosis is a potentially fatal pathological process that occurs in early brain injury (EBI) after subarachnoid hemorrhage (SAH). There is an urgent need to identify effective therapeutics to alleviate neuronal apoptosis. Tetramethylpyrazine (TMP), as an important component of the Chinese traditional medicinal herb Ligusticum wallichii, has been widely used in China to treat cerebral ischemic injury and confer neuroprotection. In the present work, we investigate whether TMP can reduce EBI following SAH in rats, specifically via inactivating the PERK/Akt signaling cascade. One hundred twenty-five male Sprague–Dawley rats were used in the present study. TMP was administered by intravenous (i.v.) injection, and the Akt inhibitor MK2206 was injected intracerebroventricularly (i.c.v.). SAH grade, neurological scores, and brain water content were measured 24 h after SAH. Neuronal apoptosis was visualized by Fluoro-Jade C (FJC) staining. Western blotting was used to measure the levels of PERK, p-PERK, eIF2α, p-eIF2α, Akt, p-Akt, Bcl-2, Bax, and cleaved caspase-3. Our results showed that TMP effectively reduced neuronal apoptosis and improved neurobehavioral deficits 24 h after SAH. Administration of TMP reduced the abundance of p-PERK and p-eIF2α. In addition, TMP increased the p-Akt level and the Bcl-2/Bax ratio and decreased the level of cleaved caspase-3. The selective Akt inhibitor MK2206 abolished the anti-apoptotic effect of TMP at 24 h after SAH. Collectively, these results indicate that Akt-related anti-apoptosis through the PERK pathway is a major, potent mechanism of EBI. Further investigation of this pathway may provide a basis for the development of TMP as a clinical treatment.

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References

  1. Lawton MT, Vates GE (2017) Subarachnoid hemorrhage. N Engl J Med 377(3):257–266. https://doi.org/10.1056/NEJMcp1605827

    Article  PubMed  Google Scholar 

  2. Chen S, Feng H, Sherchan P, Klebe D, Zhao G, Sun X, Zhang J, Tang J, Zhang JH (2014) Controversies and evolving new mechanisms in subarachnoid hemorrhage. Prog Neurobiol 115:64–91. https://doi.org/10.1016/j.pneurobio.2013.09.002

    Article  PubMed  Google Scholar 

  3. Yan F, Cao S, Li J, Dixon B, Yu X, Chen J, Gu C, Lin W, Chen G (2017) Pharmacological inhibition of PERK attenuates early brain injury after subarachnoid hemorrhage in rats through the activation of Akt. Mol Neurobiol 54(3):1808–1817. https://doi.org/10.1007/s12035-016-9790-9

    Article  PubMed  CAS  Google Scholar 

  4. Chen S, Wu H, Tang J, Zhang J, Zhang JH (2015) Neurovascular events after subarachnoid hemorrhage: focusing on subcellular organelles. Acta Neurochir Suppl 120:39–46. https://doi.org/10.1007/978-3-319-04981-6_7

    Article  PubMed  PubMed Central  Google Scholar 

  5. Zhao J, Xiang X, Zhang H, Jiang D, Liang Y, Qing W, Liu L, Zhao Q, He Z (2017) CHOP induces apoptosis by affecting brain iron metabolism in rats with subarachnoid hemorrhage. Exp Neurol 302:22–33. https://doi.org/10.1016/j.expneurol.2017.12.015

    Article  PubMed  CAS  Google Scholar 

  6. Zhu X, Wang K, Zhang K, Tan X, Wu Z, Sun S, Zhou F, Zhu L (2015) Tetramethylpyrazine protects retinal capillary endothelial cells (TR-iBRB2) against IL-1beta-induced nitrative/oxidative stress. Int J Mol Sci 16(9):21775–21790. https://doi.org/10.3390/ijms160921775

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  7. Gao C, Liu X, Liu W, Shi H, Zhao Z, Chen H, Zhao S (2008) Anti-apoptotic and neuroprotective effects of Tetramethylpyrazine following subarachnoid hemorrhage in rats. Autonomic Neurosci 141(1–2):22–30. https://doi.org/10.1016/j.autneu.2008.04.007

    Article  CAS  Google Scholar 

  8. Shao Z, Li J, Zhao Z, Gao C, Sun Z, Liu X (2010) Effects of tetramethylpyrazine on nitric oxide/cGMP signaling after cerebral vasospasm in rabbits. Brain Res 1361:67–75. https://doi.org/10.1016/j.brainres.2010.09.011

    Article  PubMed  CAS  Google Scholar 

  9. Liu W, Liu K, Zhang S, Shan L, Tang J (2018) Tetramethylpyrazine showed therapeutic effects on sepsis-induced acute lung injury in rats by inhibiting endoplasmic reticulum stress protein kinase RNA-like endoplasmic reticulum kinase (PERK) signaling-induced apoptosis of pulmonary microvascular endothelial cells. Med Sci Monit 24:1225–1231

    Article  PubMed  PubMed Central  Google Scholar 

  10. Sun X, Liao W, Wang J, Wang P, Gao H, Wang M, Xu C, Zhong Y, Ding Y (2016) CSTMP induces apoptosis and mitochondrial dysfunction in human myeloma RPMI8226 cells via CHOP-dependent endoplasmic reticulum stress. Biomed Pharmacother 83:776–784. https://doi.org/10.1016/j.biopha.2016.07.045

    Article  PubMed  CAS  Google Scholar 

  11. Wu P, Li Y, Zhu S, Wang C, Dai J, Zhang G, Zheng B, Xu S, Wang L, Zhang T, Zhou P, Zhang JH, Shi H (2017) Mdivi-1 alleviates early brain injury after experimental subarachnoid hemorrhage in rats, possibly via inhibition of Drp1-activated mitochondrial fission and oxidative stress. Neurochem Res 42(5):1449–1458. https://doi.org/10.1007/s11064-017-2201-4

    Article  PubMed  CAS  Google Scholar 

  12. Sugawara T, Ayer R, Jadhav V, Zhang JH (2008) A new grading system evaluating bleeding scale in filament perforation subarachnoid hemorrhage rat model. J Neurosci Methods 167(2):327–334. https://doi.org/10.1016/j.jneumeth.2007.08.004

    Article  PubMed  Google Scholar 

  13. Garcia JH, Wagner S, Liu KF, Hu XJ (1995) Neurological deficit and extent of neuronal necrosis attributable to middle cerebral artery occlusion in rats. Statistical validation. Stroke 26(4):627–634; discussion 635

    Article  PubMed  CAS  Google Scholar 

  14. Chen S, Ma Q, Krafft PR, Chen Y, Tang J, Zhang J, Zhang JH (2013) P2 × 7 receptor antagonism inhibits p38 mitogen-activated protein kinase activation and ameliorates neuronal apoptosis after subarachnoid hemorrhage in rats. Crit Care Med 41(12):e466–e474. https://doi.org/10.1097/CCM.0b013e31829a8246

    Article  PubMed  CAS  Google Scholar 

  15. Wang R, Ma WG, Gao GD, Mao QX, Zheng J, Sun LZ, Liu YL (2011) Fluoro jade-C staining in the assessment of brain injury after deep hypothermia circulatory arrest. Brain Res 1372:127–132. https://doi.org/10.1016/j.brainres.2010.11.059

    Article  PubMed  CAS  Google Scholar 

  16. Zhao Y, Liu Y, Chen K (2016) Mechanisms and clinical application of tetramethylpyrazine (an interesting natural compound isolated from Ligusticum wallichii): current status and perspective. Oxidative Med Cell Longev 2016:2124638. https://doi.org/10.1155/2016/2124638

    Article  CAS  Google Scholar 

  17. Zhang G, Zhang F, Zhang T, Gu J, Li C, Sun Y, Yu P, Zhang Z, Wang Y (2016) Tetramethylpyrazine nitrone improves neurobehavioral functions and confers neuroprotection on rats with traumatic brain injury. Neurochem Res 41(11):2948–2957. https://doi.org/10.1007/s11064-016-2013-y

    Article  PubMed  CAS  Google Scholar 

  18. Hu J, Lang Y, Cao Y, Zhang T, Lu H (2015) The neuroprotective effect of tetramethylpyrazine against contusive spinal cord injury by activating PGC-1alpha in rats. Neurochem Res 40(7):1393–1401. https://doi.org/10.1007/s11064-015-1606-1

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  19. Zhang C, Teng F, Tu J, Zhang D (2014) Ultrasound-enhanced protective effect of tetramethylpyrazine against cerebral ischemia/reperfusion injury. PLoS One 9(11):e113673. https://doi.org/10.1371/journal.pone.0113673

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  20. Lu C, Zhang J, Shi X, Miao S, Bi L, Zhang S, Yang Q, Zhou X, Zhang M, Xie Y, Miao Q, Wang S (2014) Neuroprotective effects of tetramethylpyrazine against dopaminergic neuron injury in a rat model of Parkinson’s disease induced by MPTP. Int J Biol Sci 10(4):350–357. https://doi.org/10.7150/ijbs.8366

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  21. Rasheva VI, Domingos PM (2009) Cellular responses to endoplasmic reticulum stress and apoptosis. Apoptosis 14(8):996–1007. https://doi.org/10.1007/s10495-009-0341-y

    Article  PubMed  Google Scholar 

  22. Roussel BD, Kruppa AJ, Miranda E, Crowther DC, Lomas DA, Marciniak SJ (2013) Endoplasmic reticulum dysfunction in neurological disease. Lancet Neurol 12(1):105–118. https://doi.org/10.1016/S1474-4422(12)70238-7

    Article  PubMed  CAS  Google Scholar 

  23. Goswami P, Gupta S, Biswas J, Sharma S, Singh S (2016) Endoplasmic reticulum stress instigates the rotenone induced oxidative apoptotic neuronal death: a study in rat brain. Mol Neurobiol 53(8):5384–5400. https://doi.org/10.1007/s12035-015-9463-0

    Article  PubMed  CAS  Google Scholar 

  24. Li H, Yu JS, Zhang HS, Yang YQ, Huang LT, Zhang DD, Hang CH (2016) Increased expression of caspase-12 after experimental subarachnoid hemorrhage. Neurochem Res 41(12):3407–3416. https://doi.org/10.1007/s11064-016-2076-9

    Article  PubMed  CAS  Google Scholar 

  25. Gharibani P, Modi J, Menzie J, Alexandrescu A, Ma Z, Tao R, Prentice H, Wu JY (2015) Comparison between single and combined post-treatment with S-Methyl-N,N-diethylthiolcarbamate sulfoxide and taurine following transient focal cerebral ischemia in rat brain. Neuroscience 300:460–473. https://doi.org/10.1016/j.neuroscience.2015.05.042

    Article  PubMed  CAS  Google Scholar 

  26. Rubovitch V, Barak S, Rachmany L, Goldstein RB, Zilberstein Y, Pick CG (2015) The neuroprotective effect of salubrinal in a mouse model of traumatic brain injury. Neuromol Med 17(1):58–70. https://doi.org/10.1007/s12017-015-8340-3

    Article  CAS  Google Scholar 

  27. Moreno JA, Halliday M, Molloy C, Radford H, Verity N, Axten JM, Ortori CA, Willis AE, Fischer PM, Barrett DA, Mallucci GR (2013) Oral treatment targeting the unfolded protein response prevents neurodegeneration and clinical disease in prion-infected mice. Sci Translational Med 5(206):206ra138. https://doi.org/10.1126/scitranslmed.3006767

    Article  CAS  Google Scholar 

  28. Xie Z, Enkhjargal B, Wu L, Zhou K, Sun C, Hu X, Gospodarev V, Tang J, You C, Zhang JH (2018) Exendin-4 attenuates neuronal death via GLP-1R/PI3K/Akt pathway in early brain injury after subarachnoid hemorrhage in rats. Neuropharmacology 128:142–151. https://doi.org/10.1016/j.neuropharm.2017.09.040

    Article  PubMed  CAS  Google Scholar 

  29. Xu H, Li J, Wang Z, Feng M, Shen Y, Cao S, Li T, Peng Y, Fan L, Chen J, Gu C, Yan F, Wang L, Chen G (2017) Methylene blue attenuates neuroinflammation after subarachnoid hemorrhage in rats through the Akt/GSK-3beta/MEF2D signaling pathway. Brain Behav Immun 65:125–139. https://doi.org/10.1016/j.bbi.2017.04.020

    Article  PubMed  CAS  Google Scholar 

  30. Yan S, Chen L, Wei X, Cheng L, Kong L, Liu X, Zhang X, Liu H (2015) Tetramethylpyrazine analogue CXC195 ameliorates cerebral ischemia-reperfusion injury by regulating endothelial nitric oxide synthase phosphorylation via PI3K/Akt signaling. Neurochem Res 40(3):446–454. https://doi.org/10.1007/s11064-014-1485-x

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

This work was supported by a grant from the Youth Science Foundation of Heilongjiang Province (QC2014C102).

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  1. Zhengkai Shao and Pei Wu have contributed equally to this work.

Authors and Affiliations

  1. Department of Neurosurgery, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, People’s Republic of China

    Zhengkai Shao, Xuefeng Wang, Meishan Jin, Shuang Liu & Xudong Ma

  2. Department of Neurosurgery, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, People’s Republic of China

    Pei Wu & Huaizhang Shi

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  1. Zhengkai Shao
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Correspondence to Huaizhang Shi.

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Shao, Z., Wu, P., Wang, X. et al. Tetramethylpyrazine Protects Against Early Brain Injury and Inhibits the PERK/Akt Pathway in a Rat Model of Subarachnoid Hemorrhage. Neurochem Res 43, 1650–1659 (2018). https://doi.org/10.1007/s11064-018-2581-0

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