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Expression of HSPA8 in Nucleus Pulposus of Lumbar Intervertebral Disc and Its Effect on Degree of Degeneration

Abstract

Introduction

This study aimed to investigate the expression of a 70-kDa heat shock protein [heat shock 70-kDa protein 8 (HSPA8)/heat shock protein 70 (Hsc70)] in human degenerative lumbar intervertebral discs and its relationship with the degree of degeneration of human intervertebral discs.

Methods

A total of 72 cases of lumbar intervertebral disc nucleus pulposus tissues were collected. Among these, 18 cases of nucleus pulposus tissue were assigned to the control group, while 54 cases of nucleus pulposus tissues were assigned to the experimental group. According to the preoperative MRI, cases in the experimental group were further divided into three groups: protrusion group (n = 18), extrusion group (n = 18), and sequestration group (n = 18). Western blot was performed to determine the relative expression of HSPA8 in the nucleus pulposus in each group. Hematoxylin and eosin staining was performed to determine the number of nucleus pulposus cells, morphological differences, and cell densities of the degenerated intervertebral discs and normal intervertebral discs. Immunohistochemistry was performed to determine the expression of HSPA8 in nucleus pulposus tissues in each group.

Results

Hematoxylin and eosin staining results: There were significant differences in cell morphology and number between the control group and the experimental group. Furthermore, there were significant differences in cell density (F = 936.80, P < 0.01). Immunohistochemistry results: HSPA8 was expressed in lumbar intervertebral disc nucleus pulposus tissues, and its expression of gradually decreased with the severity of the disease, and the differences were significant (F = 2110.43, P < 0.01). Western blot results: The expression of HSPA8 in human degenerative nucleus pulposus tissues gradually decreased, and the differences were significant (F = 1841.72, P < 0.01).

Conclusion

HSPA8 is stably expressed in human intervertebral disc nucleus pulposus tissues, and its expression is associated with the degree of intervertebral disc degeneration.

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References

  1. Feng C, Liu H, Yang Y, Huang B, Zhou Y. Growth and differentiation factor-5 contributes to the structural and functional maintenance of the intervertebral disc. Cell Physiol Biochem. 2015;35:1–16.

    Article  Google Scholar 

  2. Blanquer SB, Grijpma DW, Poot AA. Delivery systems for the treatment of degenerated intervertebral discs. Adv Drug Deliv Rev. 2015;84:172–87.

    CAS  Article  Google Scholar 

  3. Ma CJ, Luo SS, Li JS, Cao HJ, An ZW, Zhang X. Role of HIF in the pathology mechanism of the herniation of lumbar intervertebral disc. Anhui Med Pharm J. 2017;21:1306–10.

    Google Scholar 

  4. Ohashi M, Watanabe K, Hirano T, Hasegawa K, Katsumi K, Shoji H, Mizouchi T, Endo N. Predicting factors at skeletal maturity for curve progression and low back pain in adult patients treated nonoperatively for adolescent idiopathic scoliosis with thoracolumbar/lumbar curves: a mean 25-year follow-up. Spine (Phila Pa 1976). 2018;43:E1403–11.

    Article  Google Scholar 

  5. Matsui H, Kanamori M, Ishihara H, Yudoh K, Naruse Y, Tsuji H. Familial predisposition for lumbar degenerative disc disease. A case-control study. Spine (Phila Pa 1976). 1998;23:1029–34.

    CAS  Article  Google Scholar 

  6. Ala-Kokko L. Genetic risk factors for lumbar disc disease. Ann Med. 2002;34:42–7.

    CAS  Article  Google Scholar 

  7. Schistad EI, Bjorland S, Røe C, Gjerstad J, Vetti N, Myhre K, Espeland A. Five-year development of lumbar disc degeneration-a prospective study. Skelet Radiol. 2019;48:871–9.

    Article  Google Scholar 

  8. Song YQ, Karasugi T, Cheung KM, Chiba K, Ho DW, Miyake A, Kao PY, Sze KL, Yee A, Takahashi A, Kawaguchi Y, Mikami Y, Matsumoto M, Togawa D, Kanayama M, Shi D, Dai J, Jiang Q, Wu C, Tian W, Wang N, Leong JC, Luk KD, Yip SP, Cherny SS, Wang J, Mundlos S, Kelempisioti A, Eskola PJ, Männikkö M, Mäkelä P, Karppinen J, Järvelin MR, O’Reilly PF, Kubo M, Kimura T, Kubo T, Toyama Y, Mizuta H, Cheah KS, Tsunoda T, Sham PC, Ikegawa S, Chan D. Lumbar disc degeneration is linked to a carbohydrate sulfotransferase 3 variant. J Clin Investig. 2013;123:4909–17.

    CAS  Article  Google Scholar 

  9. Zhang WQ, Jiang J, Pang XD, et al. Mechanism of lumbar disc degeneration. Chin J Orthop. 2008;21:1635–6.

    Google Scholar 

  10. Yaltirik CK, Timirci-Kahraman Ö, Gulec-Yilmaz S, Ozdogan S, Atalay B, Isbir T. The evaluation of proteoglycan levels and the possible role of gene (c.6423T > C) variant in patients with lumbar disc degeneration disease. In Vivo. 2019;33:413–7.

    CAS  Article  Google Scholar 

  11. Lamian V, Small GM, Feldherr CM. Evidence for the existence of a novel mechanism for the nuclear import of Hsc70. Exp Cell Res. 1996;228:84–91.

    CAS  Article  Google Scholar 

  12. Marciniak J, Lossdörfer S, Kirschneck C, Deschner J, Jäger A, Wolf M. Heat shock protein 70 dampens the inflammatory response of human PDL cells to mechanical loading in vitro. J Periodontal Res. 2019. https://doi.org/10.1111/jre.12648.

    Article  PubMed  Google Scholar 

  13. Bi X, Xu M, Li J, Huang T, Jiang B, Shen L, Luo L, Liu S, Yin Z. Heat shock protein 27 inhibits HMGB1 translocation by regulating CBP acetyltransferase activity and ubiquitination. Mol Immunol. 2019;108:45–55.

    CAS  Article  Google Scholar 

  14. Hunt C, Morimoto RI. Conserved features of eukaryotic hsp70 genes revealed by comparison with the nucleotide sequence of human hsp70. Proc Natl Acad Sci USA. 1985;82:6455–9.

    CAS  Article  Google Scholar 

  15. Liu T, Daniels CK, Cao S. Comprehensive review on the HSC70 functions, interactions with related molecules and involvement in clinical diseases and therapeutic potential. Pharmacol Ther. 2012;136:354–74.

    CAS  Article  Google Scholar 

  16. Kityk R, Kopp J, Sinning I, Mayer MP. Structure and dynamics of the ATP-bound open conformation of Hsp70 chaperones. Mol Cell. 2012;48:863–74.

    CAS  Article  Google Scholar 

  17. Loeffler DA, Klaver AC, Coffey MP, Aasly JO, LeWitt PA. Age-related decrease in heat shock 70-kDa protein 8 in cerebrospinal fluid is associated with increased oxidative stress. Front Aging Neurosci. 2016;8:178.

    Article  Google Scholar 

  18. Ye D, Liang W, Dai L, Zhou L, Yao Y, Zhong X, Chen H, Xu J. Comparative and quantitative proteomic analysis of normal and degenerated human annulus fibrosus cells. Clin Exp Pharmacol Physiol. 2015;42:530–6.

    CAS  Article  Google Scholar 

  19. Spengler DM. Lumbar discectomy Results with limited disc excision and selective foraminotomy. Spine (Phila Pa 1976). 1982;7:604–7.

    CAS  Article  Google Scholar 

  20. Vadalà G, Russo F, Di Martino A, Denaro V. Intervertebral disc regeneration: from the degenerative cascade to molecular therapy and tissue engineering. J Tissue Eng Regen Med. 2015;9:679–90.

    Article  Google Scholar 

  21. Gornet MG, Peacock J, Claude J, Schranck FW, Copay AG, Eastlack RK, Benz R, Olshen A, Lotz JC. Magnetic resonance spectroscopy (MRS) can identify painful lumbar discs and may facilitate improved clinical outcomes of lumbar surgeries for discogenic pain. Eur Spine J. 2019;28:674–87.

    Article  Google Scholar 

  22. Zehra U, Cheung JPY, Bow C, Lu W, Samartzis D. Multidimensional vertebral endplate defects are associated with disc degeneration, Modic changes, facet joint abnormalities and pain. J Orthop Res. 2019;37:1080–9.

    CAS  Article  Google Scholar 

  23. Stricher F, Macri C, Ruff M, Muller S. HSPA8/HSC70 chaperone protein: structure, function, and chemical targeting. Autophagy. 2013;9:1937–54.

    CAS  Article  Google Scholar 

  24. Goldfarb SB, Kashlan OB, Watkins JN, Suaud L, Yan W, Kleyman TR, Rubenstein RC. Differential effects of Hsc70 and Hsp70 on the intracellular trafficking and functional expression of epithelial sodium channels. Proc Natl Acad Sci USA. 2006;103:5817–22.

    CAS  Article  Google Scholar 

  25. Deane EE, Woo NY. Impact of heavy metals and organochlorines on hsp70 and hsc70 gene expression in black sea bream fibroblasts. Aquat Toxicol. 2006;79:9–15.

    CAS  Article  Google Scholar 

  26. Geraci F, Pinsino A, Turturici G, Savona R, Giudice G, Sconzo G. Nickel, lead, and cadmium induce differential cellular responses in sea urchin embryos by activating the synthesis of different HSP70s. Biochem Biophys Res Commun. 2004;322:873–7.

    CAS  Article  Google Scholar 

  27. Makino H, Seki S, Yahara Y, Shiozawa S, Aikawa Y, Motomura H, Nogami M, Watanabe K, Sainoh T, Ito H, Tsumaki N, Kawaguchi Y, Yamazaki M, Kimura T. A selective inhibition of c-Fos/activator protein-1 as a potential therapeutic target for intervertebral disc degeneration and associated pain. Sci Rep. 2017;7:16983.

    Article  Google Scholar 

  28. Jiang PF, Ma ZW, Zhang MZ, Deng YJ. Expression of ADAMTS-7 and its mechanism in patients with degenerative changes in lumbar intervertebral disc. Pract J Clin Med. 2017;14:101–4.

    Google Scholar 

  29. Nikolaou G, Zibis AH, Fyllos AH, Katsioulis A, Sotiriou S, Kotrotsios A, Sgantzos M, Vassiou A, Arvanitis DL. Detection of O-Linked-N-Acetylglucosamine modification and its associated enzymes in human degenerated intervertebral discs. Asian Spine J. 2017;11:863–9.

    Article  Google Scholar 

  30. Johnson ZI, Gogate SS, Day R, Binch A, Markova DZ, Chiverton N, Cole A, Conner M, Shapiro IM, Le Maitre CL, Risbud MV. Aquaporin 1 and 5 expression decreases during human intervertebral disc degeneration: novel HIF-1-mediated regulation of aquaporins in NP cells. Oncotarget. 2015;6:11945–58.

    PubMed  PubMed Central  Google Scholar 

  31. Wang J, Liu X, Sun B, Du W, Zheng Y, Sun Y. Upregulated miR-154 promotes ECM degradation in intervertebral disc degeneration. J Cell Biochem. 2019. https://doi.org/10.1002/jcb.28471.

    Article  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

Funding

This study was sponsored by National Natural Science Foundation of China: Project approval number: 81371998. The Rapid Service Fee was funded by the authors.

Authorship

All named authors meet the International Committee of Medical Journal Editors (ICMJE) criteria for authorship for this article, take responsibility for the integrity of the work as a whole, and have given their approval for this version to be published.

Disclosures

Yong Liu. Hua-Cong Wang, Hong-Fei Xiang, Cang-Hai Jin and Bo-Hua Chen have nothing to disclose.

Compliance with Ethics Guidelines

This study was conducted in accordance with the Declaration of Helsinki. This study was conducted with approval from the Ethics Committee of The Affiliated Hospital of Qingdao University. Written informed consent was obtained from the participants.

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Correspondence to Bo-Hua Chen.

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Liu, Y., Wang, HC., Xiang, HF. et al. Expression of HSPA8 in Nucleus Pulposus of Lumbar Intervertebral Disc and Its Effect on Degree of Degeneration. Adv Ther 37, 390–401 (2020). https://doi.org/10.1007/s12325-019-01136-9

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  • DOI: https://doi.org/10.1007/s12325-019-01136-9

Keywords

  • Hematoxylin
  • HSC70 heat shock proteins
  • Immunohistochemistry
  • Intervertebral disc degeneration
  • Rheumatology
  • Western blotting