Skip to main content

Advertisement

SpringerLink
Log in

Shikonin inhibits neuronal apoptosis via regulating endoplasmic reticulum stress in the rat model of double-level chronic cervical cord compression

  • Original Article
  • Published:
Cell Biology and Toxicology Aims and scope Submit manuscript

Abstract

Cervical spondylotic myelopathy (CSM) is a clinically symptomatic entity arising from the spinal cord compression by degenerative diseases. Although endoplasmic reticulum (ER) stress has been commonly observed in several neurodegenerative diseases, the relationship between ER stress and CSM remains unknown. Shikonin is known to protect PC12 by inhibiting apoptosis in vitro. This study hypothesised that ER stress was vital in neuronal apoptosis in CSM. Shikonin might inhibit such responses by regulating ER stress through the protein kinase-like ER kinase-eukaryotic translation initiation factor 2 α-subunit-C/EBP homologous protein (PERK-eIF2α-CHOP) signalling pathway. Thus, the aim of this study was evaluating the neuroprotective effect of shikonin in rats with double-level chronic cervical cord compression, as well as primary rat cortical neurons with glutamate-induced neurotoxicity. The result showed that ER stress–related upregulation of PERK-eIF2α-CHOP resulted in rat neuronal apoptosis after chronic cervical cord compression; then, shikonin promoted motor recovery and inhibited neuronal apoptosis by attenuating PERK-eIF2α-CHOP and prevented Bax translocation from cytoplasm to mitochondrion induced by CHOP of neurons in rats with chronic compression. Also, it was found that shikonin could protect rat primary cortical neuron against glutamate toxicity by regulating ER stress through the PERK-eIF2α-CHOP pathway in vitro. In conclusion, shikonin might inhibit neuronal apoptosis by regulating ER stress through attenuating the activation of PERK-eIF2α-CHOP.

Graphical abstract

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
Fig. 9
Fig. 10
Fig. 11
Fig. 12
Fig. 13
Fig. 14
Fig. 15

Similar content being viewed by others

Availability of data and material

All the data were in the paper.

Code availability

Not applicable.

References

  • Aluvila S, Mandal T, Hustedt E, Fajer P, Choe JY, Oh KJ. Organization of the mitochondrial apoptotic BAK pore: oligomerization of the BAK homodimers. J Biol Chem. 2014;289:2537–51.

    Article  CAS  PubMed  Google Scholar 

  • Bakhsheshian J, Mehta VA, Liu JC. Current diagnosis and management of cervical spondylotic myelopathy. Global Spine J. 2017;7:572–86.

    Article  PubMed  PubMed Central  Google Scholar 

  • Basso DM, Beattie MS, Bresnahan JC. A sensitive and reliable locomotor rating scale for open field testing in rats. J Neurotrauma. 1995;12:1–21.

    Article  CAS  PubMed  Google Scholar 

  • Bleicken S, Classen M, Padmavathi PV, Ishikawa T, Zeth K, Steinhoff HJ, et al. Molecular details of Bax activation, oligomerization, and membrane insertion. J Biol Chem. 2010;285:6636–47.

    Article  CAS  PubMed  Google Scholar 

  • Bleicken S, Jeschke G, Stegmueller C, Salvador-Gallego R, García-Sáez AJ, Bordignon E. Structural model of active Bax at the membrane. Mol Cell. 2014;56:496–505.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bi Y, Zhu Y, Zhang M, Zhang K, Hua X, Fang Z, et al. Effect of shikonin on spinal cord injury in rats via regulation of HMGB1/TLR4/NF-kB signaling pathway. Cell Physiol Biochem. 2017;43:481–91.

    Article  CAS  PubMed  Google Scholar 

  • Chen B, Li Y, Yu B, Zhang Z, Brommer B, Williams PR, Liu Y, Hegarty SV, Zhou S, Zhu J, Guo H, Lu Y, Zhang Y, Gu X, He Z. Reactivation of Dormant Relay Pathways in Injured Spinal Cord by KCC2 Manipulations. Cell. 2018;174(3):521–35.e13.

  • Czabotar PE, Westphal D, Dewson G, Ma S, Hockings C, Fairlie WD, et al. Bax crystal structures reveal how BH3 domains activate Bax and nucleate its oligomerization to induce apoptosis. Cell. 2013;152:519–31.

    Article  CAS  PubMed  Google Scholar 

  • Dewson G, Ma S, Frederick P, Hockings C, Tan I, Kratina T, et al. Bax dimerizes via a symmetric BH3:groove interface during apoptosis. Cell Death Differ. 2012;19:661–70.

    Article  CAS  PubMed  Google Scholar 

  • Ducourneau VRR, Dolique T, Hachem-Delaunay S, Miraucourt LS, Amadio A, Blaszczyk L, et al. Cancer pain is not necessarily correlated with spinal overexpression of reactive glia markers. Pain. 2014;155:275–91.

    Article  CAS  PubMed  Google Scholar 

  • Errea O, Moreno B, Gonzalez-Franquesa A, Garcia-Roves PM, Villoslada P. The disruption of mitochondrial axonal transport is an early event in neuroinflammation. J Neuroinflammation. 2015;12:152.

    Article  PubMed  PubMed Central  Google Scholar 

  • Esmaeilzadeh E, Gardaneh M, Gharib E, Sabouni F. Shikonin protects dopaminergic cell line PC12 against 6-hydroxydopamine-mediated neurotoxicity via both glutathione-dependent and independent pathways and by inhibiting apoptosis. Neurochem Res. 2013;38:1590–604.

    Article  CAS  PubMed  Google Scholar 

  • Gara RK, Srivastava VK, Duggal S, Bagga JK, Bhatt M, Sanyal S, et al. Shikonin selectively induces apoptosis in human prostate cancer cells through the endoplasmic reticulum stress and mitochondrial apoptotic pathway. J Biomed Sci. 2015;22:26.

    Article  PubMed  PubMed Central  Google Scholar 

  • Gotoh T, Terada K, Oyadomari S, Mori M. hsp70-DnaJ chaperone pair prevents nitric oxide- and CHOP-induced apoptosis by inhibiting translocation of Bax to mitochondria. Cell Death Differ. 2004;11:390–402.

    Article  CAS  PubMed  Google Scholar 

  • Gerakis Y, Hetz C. Emerging roles of ER stress in the etiology and pathogenesis of Alzheimer’s disease. FEBS J. 2018;285:995–1011.

  • Green DR, Galluzzi L, Kroemer G. Metabolic control of cell death. Science. 2014;345:1250256.

    Article  PubMed  PubMed Central  Google Scholar 

  • Guo C, He J, Song X, Tan L, Wang M, Jiang P, et al. Pharmacological properties and derivatives of shikonin-A review in recent years. Pharmacol Res. 2019;149:104463.

    Article  CAS  PubMed  Google Scholar 

  • Gorman AM, Healy SJ, Jäger R, Samali A. Stress management at the ER: regulators of ER stress-induced apoptosis. Pharmacol Ther. 2012;134:306–16.

    Article  CAS  PubMed  Google Scholar 

  • Hassannejad Z, Zadegan SA, Shakouri-Motlagh A, Mokhatab M, Rezvan M, Sharif-Alhoseini M, et al. The fate of neurons after traumatic spinal cord injury in rats: a systematic review. Iran J Basic Med Sci. 2018;21:546–57.

    PubMed  PubMed Central  Google Scholar 

  • He B, Zhang W, Qiao J, Peng Z, Chai X. Melatonin protects against COPD by attenuating apoptosis and endoplasmic reticulum stress via upregulating SIRT1 expression in rats. Can J Physiol Pharmacol. 2019;97:386–91.

    Article  CAS  PubMed  Google Scholar 

  • Hetz C, Mollereau B. Disturbance of endoplasmic reticulum proteostasis in neurodegenerative diseases. Nat Rev Neurosci. 2014;15:233–49.

    Article  CAS  PubMed  Google Scholar 

  • Hetz C, Saxena S. ER stress and the unfolded protein response in neurodegeneration. Nat Rev Neurol. 2017;13:477–91.

    Article  CAS  PubMed  Google Scholar 

  • Hong D, Jang SY, Jang EH, Jung B, Cho IH, Park MJ, et al. Shikonin as an inhibitor of the LPS-induced epithelial-to-mesenchymal transition in human breast cancer cells. Int J Mol Med. 2015;36:1601–6.

    Article  CAS  PubMed  Google Scholar 

  • Hu H, Tian M, Ding C, Yu S. The C/EBP homologous protein (CHOP) transcription factor functions in endoplasmic reticulum stress-induced apoptosis and microbial infection. Front Immunol. 2018;9:3083.

    Article  CAS  PubMed  Google Scholar 

  • Iyer A, Azad TD, Tharin S. Cervical spondylotic myelopathy. . Clin Spine Surg. 2016;29:408–14.

    Article  PubMed  Google Scholar 

  • Kamarehei M, Kabudanian Ardestani S, Firouzi M, Zahednasab H, Keyvani H, Harirchian MH. Increased expression of endoplasmic reticulum stress-related caspase-12 and CHOP in the hippocampus of EAE mice. Brain Res Bull. 2019;147:174–82.

    Article  CAS  PubMed  Google Scholar 

  • Karadimas SK, Gatzounis G, Fehlings MG. Pathobiology of cervical spondylotic myelopathy. Eur Spine J. 2015a;24(Suppl 2):132–8.

    Article  PubMed  Google Scholar 

  • Karadimas SK, Gatzounis G, Fehlings MG. Pathobiology of cervical spondylotic myelopathy. Eur Spine J. 2015b;24:132–8.

    Article  PubMed  Google Scholar 

  • Lebl DR, Hughes A, Cammisa FP, O’Leary PF. Cervical spondylotic myelopathy: pathophysiology, clinical presentation, and treatment. HSS J. 2011;7:170–8.

    Article  PubMed  PubMed Central  Google Scholar 

  • Li H, Zhang X, Qi X, Zhu X, Cheng L. Icariin inhibits endoplasmic reticulum stress-induced neuronal apoptosis after spinal cord injury through modulating the PI3K/AKT signaling pathway. Int J Biol Sci. 2019;15:277–86.

    Article  PubMed  PubMed Central  Google Scholar 

  • Li X, Yu X, Zhou D, Chen B, Li W, Zheng X, et al. CCT020312 inhibits triple-negative breast cancer through PERK pathway-mediated G1 phase cell cycle arrest and apoptosis. Front Pharmacol. 2020;11:737.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu D, Zhang M, Yin H. Signaling pathways involved in endoplasmic reticulum stress-induced neuronal apoptosis. Int J Neurosci. 2013;123:155–62.

    Article  CAS  PubMed  Google Scholar 

  • Liu T, Zhang Q, Mo W, Yu Q, Xu S, Li J, et al. The protective effects of shikonin on hepatic ischemia/reperfusion injury are mediated by the activation of the PI3K/Akt pathway. Sci Rep. 2017;7:44785.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu Y, Kang X, Niu G, He S, Zhang T, Bai Y, et al. Shikonin induces apoptosis and prosurvival autophagy in human melanoma A375 cells via ROS-mediated ER stress and p38 pathways. Artif Cells Nanomed Biotechnol. 2019;47:626–35.

    Article  CAS  PubMed  Google Scholar 

  • Lu PJ, Yang C, Lin CN, Li CF, Chu CC, Wang JJ, et al. Shiunko and acetylshikonin promote reepithelialization, angiogenesis, and granulation tissue formation in wounded skin. Am J Chin Med. 2008;36:115–23.

    Article  CAS  PubMed  Google Scholar 

  • Matz PG, Anderson PA, Holly LT, Groff MW, Heary RF, Kaiser MG, et al. The natural history of cervical spondylotic myelopathy. J Neurosurg Spine. 2009;11:104–11.

    Article  PubMed  Google Scholar 

  • McCormick JR, Sama AJ, Schiller NC, Butler AJ, Donnally CJ 3rd. Cervical spondylotic myelopathy: a guide to diagnosis and management. J Am Board Fam Med. 2020;33:303–13.

    Article  PubMed  Google Scholar 

  • Nouri A, Tetreault L, Singh A, Karadimas SK, Fehlings MG. Degenerative cervical myelopathy: epidemiology, genetics, and pathogenesis. Spine (Phila Pa 1976). 2015;40:E675–93.

  • Peña-Blanco A, García-Sáez AJ. Bax, Bak and beyond-mitochondrial performance in apoptosis. FEBS J. 2018;285:416–31.

    Article  PubMed  Google Scholar 

  • Prentice H, Modi JP, Wu JY. Mechanisms of neuronal protection against excitotoxicity, endoplasmic reticulum stress, and mitochondrial dysfunction in stroke and neurodegenerative diseases. Oxid Med Cell Longev. 2015;2015:964518.

    Article  PubMed  PubMed Central  Google Scholar 

  • Rivlin AS, Tator CH. Effect of duration of acute spinal cord compression in a new acute cord injury model in the rat. Surg Neurol. 1978;10:38–43.

    CAS  PubMed  Google Scholar 

  • Rodemer W, Selzer ME. Role of axon resealing in retrograde neuronal death and regeneration after spinal cord injury. Neural Regen Res. 2019;14:399–404.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Subburaj Y, Cosentino K, Axmann M, Pedrueza-Villalmanzo E, Hermann E, Bleicken S, et al. Bax monomers form dimer units in the membrane that further self-assemble into multiple oligomeric species. Nat Commun. 2015;6:8042.

    Article  CAS  PubMed  Google Scholar 

  • Sprenkle NT, Sims SG, Sánchez CL, Meares GP. Endoplasmic reticulum stress and inflammation in the central nervous system. Mol Neurodegener. 2017;12:42.

    Article  PubMed  PubMed Central  Google Scholar 

  • Tong Y, Bai L, Gong R, Chuan J, Duan X, Zhu Y. Shikonin protects PC12 cells against beta-amyloid peptide-induced cell injury through antioxidant and antiapoptotic activities. Sci Rep. 2018;8:26.

    Article  PubMed  PubMed Central  Google Scholar 

  • Walter P, Ron D. The unfolded protein response: from stress pathway to homeostatic regulation. Science. 2011;334:1081–6.

    Article  CAS  PubMed  Google Scholar 

  • Wang L, Gai P, Xu R, Zheng Y, Lv S, Li Y, et al. Shikonin protects chondrocytes from interleukin-1beta-induced apoptosis by regulating PI3K/Akt signaling pathway. Int J Clin Exp Pathol. 2015;8:298–308.

    PubMed  PubMed Central  Google Scholar 

  • Wang X. The expanding role of mitochondria in apoptosis. Genes Dev. 2001;15:2922–33.

    CAS  PubMed  Google Scholar 

  • Xiang C, Wang Y, Zhang H, Han F. The role of endoplasmic reticulum stress in neurodegenerative disease. Apoptosis. 2017;22:1–26.

    Article  CAS  PubMed  Google Scholar 

  • Xue B, Huang J, Ma B, Yang B, Chang D, Liu J. Astragaloside IV protects primary cerebral cortical neurons from oxygen and glucose deprivation/reoxygenation by activating the PKA/CREB pathway. Neuroscience. 2019;404:326–37.

    Article  CAS  PubMed  Google Scholar 

  • Yang X, Lu F, Li L, Li J, Luo J, Zhang S, et al. Wu-Mei-wan protects pancreatic β cells by inhibiting NLRP3 inflammasome activation in diabetic mice. BMC Complement Altern Med. 2019;19:35.

    Article  PubMed  PubMed Central  Google Scholar 

  • Yang J, Wang Z, Chen DL. Shikonin ameliorates isoproterenol (ISO)-induced myocardial damage through suppressing fibrosis, inflammation, apoptosis and ER stress. Biomed Pharmacother. 2017;93:1343–57.

    Article  CAS  PubMed  Google Scholar 

  • Yoshida LS, Kakegawa T, Yuda Y, Takano-Ohmuro H. Shikonin changes the lipopolysaccharide-induced expression of inflammation-related genes in macrophages. J Nat Med. 2017;71:723–34.

    Article  CAS  PubMed  Google Scholar 

  • Zhang X, Yuan Y, Jiang L, Zhang J, Gao J, Shen Z, et al. Endoplasmic reticulum stress induced by tunicamycin and thapsigargin protects against transient ischemic brain injury: Involvement of PARK2-dependent mitophagy. Autophagy. 2014;10(10):1801–13.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhong J, Wang Z, Xie Q, Li T, Chen K, Zhu T, et al. Shikonin ameliorates D-galactose-induced oxidative stress and cognitive impairment in mice via the MAPK and nuclear factor-kappaB signaling pathway. Int Immunopharmacol. 2020;83:106491.

    Article  CAS  PubMed  Google Scholar 

  • Zhou LY, Yao M, Tian ZR, Liu SF, Song YJ, Ye J, et al. Muscone suppresses inflammatory responses and neuronal damage in a rat model of cervical spondylotic myelopathy by regulating Drp1-dependent mitochondrial fission. J Neurochem. 2020a;155:154–76.

    Article  CAS  PubMed  Google Scholar 

  • Zhou L, Yao M, Tian Z, Song Y, Sun Y, Ye J, et al. Echinacoside attenuates inflammatory response in a rat model of cervical spondylotic myelopathy via inhibition of excessive mitochondrial fission. Free Radic Biol Med. 2020b;152:697–714.

    Article  CAS  PubMed  Google Scholar 

Download references

Funding

We thank Natural Science Foundation of China (No. 82074454 for Xue-jun Cui, No. 821751915 for Min Yao, No. 81930116 for Yong-jun Wang, and No. 81873317 for Xue-jun Cui), National key R and D plan (No. 2018YFC1704302 for Yong-jun Wang), and Shanghai Natural Science Foundation (No. 20ZR1459000 for Min Yao) for their financial support.

Author information

Author notes

  1. Min Yao and Gan Li contributed equally to this work

Authors and Affiliations

  1. Spine Disease Institute, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, 725 South Wanping Road, Shanghai, 200032, China

    Min Yao, Gan Li, Zhong Zheng, Yue-li Sun, Shu-fen Liu, Yong-jun Wang & Xue-jun Cui

  2. Key Laboratory of Theory and Therapy of Muscles and Bones, Ministry of Education, Shanghai University of Traditional Chinese Medicine, Shanghai, 200032, China

    Min Yao, Gan Li, Zhong Zheng, Yue-li Sun, Shu-fen Liu, Yong-jun Wang & Xue-jun Cui

  3. Rehabilitation Medicine Center, Jiangsu Provincial People’s Hospital, Jiangsu, 210029, China

    Long-yun Zhou

Authors
  1. Min Yao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gan Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Long-yun Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Zhong Zheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yue-li Sun
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Shu-fen Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yong-jun Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Xue-jun Cui
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Min Yao, Yong-jun Wang, and Xue-jun Cui contributed to study design, data analysis, and manuscript modification. Gan Li, Long-yun Zhou, and Zhong Zheng contributed to animal model and laboratory measurements (immunohistochemistry). Min Yao, Yue-li Sun, and Shu-fen Liu contributed to in vitro cell experiments. All authors were involved in the analysis of results and manuscript preparation and approval.

Corresponding authors

Correspondence to Yong-jun Wang or Xue-jun Cui.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Ethics approval

The study was approved by the Animal Ethics Committee, Fudan University (Approval number: 202005006Z, Shanghai, China). The experimental protocol was performed according to the Guide for the Care and Use of Laboratory Animals from the National Institutes of Health (NIH Publications No. 8023).

Consent to participate

Not applicable.

Consent for publication

Not applicable.

Additional information

Publisher's note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yao, M., Li, G., Zhou, Ly. et al. Shikonin inhibits neuronal apoptosis via regulating endoplasmic reticulum stress in the rat model of double-level chronic cervical cord compression. Cell Biol Toxicol 39, 907–928 (2023). https://doi.org/10.1007/s10565-021-09648-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10565-021-09648-3

Keywords

Search

Navigation