The role and mechanisms of action of SIRT6 in the suppression of postoperative epidural scar formation

Fan,Xiaochen; Chen,Junjie; Shi,Da; Jia,Jing; He,Jia; Li,Liang; Lei,Tao; Chen,Xuesong

doi:10.3892/ijmm.2016.2522

The role and mechanisms of action of SIRT6 in the suppression of postoperative epidural scar formation

Authors:
- Xiaochen Fan
- Junjie Chen
- Da Shi
- Jing Jia
- Jia He
- Liang Li
- Tao Lei
- Xuesong Chen
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Affiliations: Department of Traditional Chinese Medicine Orthopedics Diagnosis and Treatment Center, Honghui Hospital, Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi 710054, P.R. China, Department of Orthopedics and Traumatology, The First Affiliated Hospital of Zhejiang Chinese Medicine University, Hangzhou, Zhejiang 310006, P.R. China
Published online on: March 11, 2016 https://doi.org/10.3892/ijmm.2016.2522
Pages: 1337-1344

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Abstract

In order to investigate the role which sirtuin-6 (SIRT6) plays in lumbar spinal epidural fibrosis (EF) and scar formation in vitro and in vivo, SIRT6 and transforming growth factor β (TGF‑β) protein levels in the lumbar disc of patients were detected using western blotting in patients who had undergone a laminectomy. The results demonstrated that SIRT6 expression was significantly reduced in the lumbar discs of patients in whom an epidural scar had formed, but the expression pattern of TGF-β2 was much higher. Subsequently, a pcDNA‑SIRT6 expression vector was constructed and transfected into the primary fibroblasts isolated from the epidural scars. Flow cytometric and MTT analyses indicated that overexpression of SIRT6 suppressed the proliferation of the fibroblasts, and TGF‑β2 and interleukin-1α expression, as well as collagen type I (Col I) production. The results of bioinformatics and molecular biological analyses demonstarted that TGF‑β2 was a target of microRNA-21 (miR-21) and SIRT6 overexpression suppressed the levels of TGF-β2 through promoting the expression of miR-21. Finally, by injecting the pcDNA-SIRT6 vector, it was possible to observe that SIRT6 suppressed EF and epidural scar formation in vivo. In conclusion, we noted that SIRT6 suppressed EF and epidural scar formation in vitro and in vivo. It was also noted that SIRT6 overexpression suppressed TGF-β2 levels by promoting the expression of miR-21.

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1	Alkalay RN, Kim DH, Urry DW, Xu J, Parker TM and Glazer PA: Prevention of postlaminectomy epidural fibrosis using bioelastic materials. Spine. 28:1659–1665. 2003. View Article : Google Scholar : PubMed/NCBI
2	Li C, Wang H, Liu H, Yin J, Cui L and Chen Z: The prevention effect of poly (L-glutamic acid)/chitosan on spinal epidural fibrosis and peridural adhesion in the post-laminectomy rabbit model. Eur Spine J. 23:2423–2431. 2014. View Article : Google Scholar : PubMed/NCBI
3	Zhang C, Kong X, Ning G, Liang Z, Qu T, Chen F, Cao D, Wang T, Sharma HS and Feng S: All-trans retinoic acid prevents epidural fibrosis through NF-κB signaling pathway in post-laminectomy rats. Neuropharmacology. 79:275–281. 2014. View Article : Google Scholar
4	Sharma Manoj, Dahima Rashmi and Gupta Anil Kumar: A review on role of peridural fibrosis and its diagnosis. Journal of Biomedical and Pharmaceutical Research. 2:81–87. 2013.
5	Spiegelberg L, Swagemakers SM, Van Ijcken WF, Oole E, Wolvius EB, Essers J and Braks JA: Gene expression analysis reveals inhibition of radiation-induced TGFβ-signaling by hyperbaric oxygen therapy in mouse salivary glands. Mol Med. 20:257–269. 2014. View Article : Google Scholar : PubMed/NCBI
6	Sun Y, Yan LQ, Liang Y, Li XL, Cao XJ and Lu C: Reduction of epidural scar adhesion by topical application of simvastatin after laminectomy in rats. Eur Rev Med Pharmacol Sci. 19:3–8. 2015.PubMed/NCBI
7	Wang Z, Wang Y, Xie P, Liu W and Zhang S: Calcium channel blockers in reduction of epidural fibrosis and dural adhesions in laminectomy rats. Eur J Orthop Surg Traumatol. 24(Suppl 1): S293–S298. 2014. View Article : Google Scholar
8	Chang H, Brown CW and Matzuk MM: Genetic analysis of the mammalian transforming growth factor-β superfamily. Endocr Rev. 23:787–823. 2013. View Article : Google Scholar
9	Yan J, Zhang H, Yin Y, Li J, Tang Y, Purkayastha S, Li L and Cai D: Obesity- and aging-induced excess of central transforming growth factor-β potentiates diabetic development via an RNA stress response. Nat Med. 20:1001–1008. 2014. View Article : Google Scholar : PubMed/NCBI
10	Avouac J, Palumbo K, Tomcik M, Zerr P, Dees C, Horn A, Maurer B, Akhmetshina A, Beyer C, Sadowski A, et al: Inhibition of activator protein 1 signaling abrogates transforming growth factor β-mediated activation of fibroblasts and prevents experimental fibrosis. Arthritis Rheum. 64:1642–1652. 2012. View Article : Google Scholar
11	Bhattacharyya S, Kelley K, Melichian DS, Tamaki Z, Fang F, Su Y, Feng G, Pope RM, Budinger GR, Mutlu GM, et al: Toll-like receptor 4 signaling augments transforming growth factor-β responses: a novel mechanism for maintaining and amplifying fibrosis in scleroderma. Am J Pathol. 182:192–205. 2013. View Article : Google Scholar :
12	Heldin CH and Moustakas A: Role of Smads in TGFβ signaling. Cell Tissue Res. 347:21–36. 2012. View Article : Google Scholar
13	Huang C and Ogawa R: Fibroproliferative disorders and their mechanobiology. Connect Tissue Res. 53:187–196. 2012. View Article : Google Scholar : PubMed/NCBI
14	Zgheib C, Xu J and Liechty KW: Targeting inflammatory cytokines and extracellular matrix composition to promote wound regeneration. Adv Wound Care (New Rochelle). 3:344–355. 2014. View Article : Google Scholar
15	Joko M, Osuka K, Usuda N, Atsuzawa K, Aoyama M and Takayasu M: Different modifications of phosphorylated Smad3C and Smad3L through TGF-β after spinal cord injury in mice. Neurosci Lett. 549:168–172. 2013. View Article : Google Scholar : PubMed/NCBI
16	Lagord C, Berry M and Logan A: Expression of TGFβ2 but not TGFβ1 correlates with the deposition of scar tissue in the lesioned spinal cord. Mol Cell Neurosci. 20:69–92. 2002. View Article : Google Scholar : PubMed/NCBI
17	Lin H: Sirtuins and novel protein post translational modifications. The FASEB Journal. 29:496.12015.
18	Lombard DB, Schwer B, Alt FW and Mostoslavsky R: SIRT6 in DNA repair, metabolism and ageing. J Intern Med. 263:128–141. 2008. View Article : Google Scholar : PubMed/NCBI
19	Van Meter M, Mao Z, Gorbunova V and Seluanov A: SIRT6 overexpression induces massive apoptosis in cancer cells but not in normal cells. Cell Cycle. 10:3153–3158. 2011. View Article : Google Scholar : PubMed/NCBI
20	Schwer B, Schumacher B, Lombard DB, Xiao C, Kurtev MV, Gao J, Schneider JI, Chai H, Bronson RT, Tsai LH, et al: Neural sirtuin 6 (Sirt6) ablation attenuates somatic growth and causes obesity. Proc Natl Acad Sci USA. 107:21790–21794. 2010. View Article : Google Scholar : PubMed/NCBI
21	Etchegaray JP, Zhong L and Mostoslavsky R: The histone deacetylase SIRT6: at the crossroads between epigenetics, metabolism and disease. Curr Top Med Chem. 13:2991–3000. 2013. View Article : Google Scholar : PubMed/NCBI
22	Shi K, Wang D, Cao X and Ge Y: Endoplasmic reticulum stress signaling is involved in mitomycin C (MMC)-induced apoptosis in human fibroblasts via PERK pathway. PLoS One. 8:e593302013. View Article : Google Scholar : PubMed/NCBI
23	Sekiguchi M, Konno S and Kikuchi S: The effects of a 5-HT2A receptor antagonist on blood flow in lumbar disc herniation: application of nucleus pulposus in a canine model. Eur Spine J. 17:307–313. 2008. View Article : Google Scholar
24	Verrecchia F and Mauviel A: Transforming growth factor-beta and fibrosis. World J Gastroenterol. 13:3056–3062. 2007.PubMed/NCBI
25	Kamath VV, Krishnamurthy S, Satelur KP and Rajkumar K: Transforming growth factor-β1 and TGF-β2 act synergistically in the fibrotic pathway in oral submucous fibrosis: an immunohistochemical observation. Indian J Med Paediatr Oncol. 36:111–116. 2015. View Article : Google Scholar : PubMed/NCBI
26	Feng Z, Li R, Shi H, Bi W, Hou W and Zhang X: Combined silencing of TGF-β2 and Snail genes inhibit epithelial-mesenchymal transition of retinal pigment epithelial cells under hypoxia. Graefes Arch Clin Exp Ophthalmol. 253:875–884. 2015. View Article : Google Scholar : PubMed/NCBI
27	Xiao C, Wang R-H, Lahusen TJ, Park O, Bertola A, Maruyama T, Reynolds D, Chen Q, Xu X, Young HA, et al: Progression of chronic liver inflammation and fibrosis driven by activation of c-JUN signaling in Sirt6 mutant mice. J Biol Chem. 287:41903–41913. 2012. View Article : Google Scholar : PubMed/NCBI
28	Swierczynski S, Klieser E, Illig R, Alinger-Scharinger B, Kiesslich T and Neureiter D: Histone deacetylation meets miRNA: epigenetics and post-transcriptional regulation in cancer and chronic diseases. Expert Opin Biol Ther. 15:651–664. 2015. View Article : Google Scholar : PubMed/NCBI
29	Pillai VB, Sundaresan NR and Gupta MP: Regulation of Akt signaling by sirtuins: its implication in cardiac hypertrophy and aging. Circ Res. 114:368–378. 2014. View Article : Google Scholar : PubMed/NCBI
30	Zhang P, Tu B, Wang H, Cao Z, Tang M, Zhang C, Gu B, Li Z, Wang L, Yang Y, et al: Tumor suppressor p53 cooperates with SIRT6 to regulate gluconeogenesis by promoting FoxO1 nuclear exclusion. Proc Natl Acad Sci USA. 111:10684–10689. 2014. View Article : Google Scholar : PubMed/NCBI
31	Iwashina T, Mochida J, Sakai D, Yamamoto Y, Miyazaki T, Ando K and Hotta T: Feasibility of using a human nucleus pulposus cell line as a cell source in cell transplantation therapy for intervertebral disc degeneration. Spine. 31:1177–1186. 2006. View Article : Google Scholar : PubMed/NCBI