Aquaporin 3 protects against lumbar intervertebral disc degeneration via the Wnt/β-catenin pathway

Xie,Huanxin; Jing,Yongbin; Xia,Jingjun; Wang,Xintao; You,Changcheng; Yan,Jinglong

doi:10.3892/ijmm.2016.2470

Aquaporin 3 protects against lumbar intervertebral disc degeneration via the Wnt/β-catenin pathway

Authors:
- Huanxin Xie
- Yongbin Jing
- Jingjun Xia
- Xintao Wang
- Changcheng You
- Jinglong Yan
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Affiliations: Department of Orthopedic Surgery, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China
Published online on: January 27, 2016 https://doi.org/10.3892/ijmm.2016.2470
Pages: 859-864

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Abstract

Previous studies have demonstrated that the expression of aquaporin 3 (AQP3), a water channel which promotes glycerol permeability and water transport across cell membranes, is reduced in degenerative lumbar intervertebral disc (IVD) tissues. However, the role of AQP3 in the pathogenesis of IVD degeneration has not recieved much scholarly attention. The objective of the present study was to investigate the effect of AQP3 on cell proliferation and extracellular matrix (ECM) degradation in human nucleus pulposus cells (hNPCs) using gain-of-function and loss-of-function experiments, and to determine whether Wnt/β-catenin signaling is involved in the effect of AQP3 on IVD degeneration. hNPCs were transfected with the AQP3-pcDNA3.1 plasmid or AQP3 siRNA to overexpress or suppress AQP3. An MTT assay was performed to determine cell proliferation, and we found that AQP3 promoted hNPC proliferation. The expression of aggrecan, a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)4 and ADAMTS5 was detected using western blot analysis, to examine the effect of AQP3 on ECM degradation in hNPCs. The results revealed that AQP3 inhibited ECM degradation in hNPCs. In addition, we found that Wnt/β-catenin signaling was suppressed by AQP3. However, the effect of AQP3 on hNPC proliferation and ECM degradation was reversed by treatment with lithium chloride, a known activator of Wnt/β‑catenin signaling. In conclusion, using in vitro and in vivo tests, we have reported for the first time, to the best of our knowledge, that AQP3 exerts protective effects against IVD degeneration, and these are effected, at least partially, through the inhibition of Wnt/β-catenin signaling.

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1	Podichetty VK: The aging spine: the role of inflammatory mediators in intervertebral disc degeneration. Cell Mol Biol (Noisy-le-grand). 53:4–18. 2007.
2	Speed C: Low back pain. BMJ. 328:1119–1121. 2004. View Article : Google Scholar : PubMed/NCBI
3	Takahashi K, Aoki Y and Ohtori S: Resolving discogenic pain. Eur Spine J. 17(Suppl 4): 428–431. 2008. View Article : Google Scholar : PubMed/NCBI
4	Weber KT, Jacobsen TD, Maidhof R, Virojanapa J, Overby C, Bloom O, Quraishi S, Levine M and Chahine NO: Developments in intervertebral disc disease research: pathophysiology, mechanobiology, and therapeutics. Curr Rev Musculoskelet Med. 8:18–31. 2015. View Article : Google Scholar : PubMed/NCBI
5	Wang SZ, Rui YF, Lu J and Wang C: Cell and molecular biology of intervertebral disc degeneration: current understanding and implications for potential therapeutic strategies. Cell Prolif. 47:381–390. 2014. View Article : Google Scholar : PubMed/NCBI
6	Isherwood I, Prendergast DJ, Hickey DS and Jenkins JP: Quantitative analysis of intervertebral disc structure. Acta Radiol Suppl. 369:492–495. 1986.PubMed/NCBI
7	Agre P: The aquaporin water channels. Proc Am Thorac Soc. 3:5–13. 2006. View Article : Google Scholar : PubMed/NCBI
8	Zeuthen T and Klaerke DA: Transport of water and glycerol in aquaporin 3 is gated by H⁺. J Biol Chem. 274:21631–21636. 1999. View Article : Google Scholar : PubMed/NCBI
9	Taş U, Caylı S, Inanır A, Ozyurt B, Ocaklı S, Karaca Zİ and Sarsılmaz M: Aquaporin-1 and aquaporin-3 expressions in the intervertebral disc of rats with aging. Balkan Med J. 29:349–353. 2012.
10	Li SB, Yang KS and Zhang YT: Expression of aquaporins 1 and 3 in degenerative tissue of the lumbar intervertebral disc. Genet Mol Res. 13:8225–8233. 2014. View Article : Google Scholar : PubMed/NCBI
11	Ye S, Wang J, Yang S, Xu W, Xie M, Han K, Zhang B and Wu Z: Specific inhibitory protein Dkk-1 blocking Wnt/β-catenin signaling pathway improve protectives effect on the extracellular matrix. J Huazhong Univ Sci Technolog Med Sci. 31:657–662. 2011. View Article : Google Scholar : PubMed/NCBI
12	Hiyama A, Yokoyama K, Nukaga T, Sakai D and Mochida J: A complex interaction between Wnt signaling and TNF-α in nucleus pulposus cells. Arthritis Res Ther. 15:R1892013. View Article : Google Scholar
13	Kong J, Ma X, Wang T, Ma J, Tian P, Han C, Zang J, Li P and Jiang H: Research progress of Wnt/beta-catenin and nuclear factor-kappa B pathways and their relevance to intervertebral disc degeneration. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 27:1523–1528. 2013.In Chinese.
14	Hiyama A, Sakai D, Arai F, Nakajima D, Yokoyama K and Mochida J: Effects of a glycogen synthase kinase-3β inhibitor (LiCl) on c-myc protein in intervertebral disc cells. J Cell Biochem. 112:2974–2986. 2011. View Article : Google Scholar : PubMed/NCBI
15	Hiyama A, Sakai D, Tanaka M, Arai F, Nakajima D, Abe K and Mochida J: The relationship between the Wnt/β-catenin and TGF-β/BMP signals in the intervertebral disc cell. J Cell Physiol. 226:1139–1148. 2011. View Article : Google Scholar
16	Hiyama A, Sakai D, Risbud MV, Tanaka M, Arai F, Abe K and Mochida J: Enhancement of intervertebral disc cell senescence by WNT/β-catenin signaling-induced matrix metalloproteinase expression. Arthritis Rheum. 62:3036–3047. 2010. View Article : Google Scholar : PubMed/NCBI
17	Mobasheri A, Wray S and Marples D: Distribution of AQP2 and AQP3 water channels in human tissue microarrays. J Mol Histol. 36:1–14. 2005. View Article : Google Scholar : PubMed/NCBI
18	Hagiwara K, Shinozaki T, Matsuzaki T, Takata K and Takagishi K: Immunolocalization of water channel aquaporins in human knee articular cartilage with intact and early degenerative regions. Med Mol Morphol. 46:104–108. 2013. View Article : Google Scholar : PubMed/NCBI
19	Meng JH, Ma XC, Li ZM and Wu DC: Aquaporin-1 and aquaporin-3 expressions in the temporo-mandibular joint condylar cartilage after an experimentally induced osteoarthritis. Chin Med J (Engl). 120:2191–2194. 2007.
20	Shen L, Zhu Z, Huang Y, Shu Y, Sun M, Xu H, Zhang G, Guo R, Wei W and Wu W: Expression profile of multiple aquaporins in human gastric carcinoma and its clinical significance. Biomed Pharmacother. 64:313–318. 2010. View Article : Google Scholar : PubMed/NCBI
21	Sive JI, Baird P, Jeziorsk M, Watkins A, Hoyland JA and Freemont AJ: Expression of chondrocyte markers by cells of normal and degenerate intervertebral discs. Mol Pathol. 55:91–97. 2002. View Article : Google Scholar : PubMed/NCBI
22	Roughley PJ, Alini M and Antoniou J: The role of proteoglycans in aging, degeneration and repair of the intervertebral disc. Biochem Soc Trans. 30:869–874. 2002. View Article : Google Scholar : PubMed/NCBI
23	David G, Ciurea AV, Mitrica M and Mohan A: Impact of changes in extracellular matrix in the lumbar degenerative disc. J Med Life. 4:269–274. 2011.
24	Le Maitre CL, Pockert A, Buttle DJ, Freemont AJ and Hoyland JA: Matrix synthesis and degradation in human intervertebral disc degeneration. Biochem Soc Trans. 35:652–655. 2007. View Article : Google Scholar : PubMed/NCBI
25	Tian Y, Yuan W, Fujita N, Wang J, Wang H, Shapiro IM and Risbud MV: Inflammatory cytokines associated with degenerative disc disease control aggrecanase-1 (ADAMTS-4) expression in nucleus pulposus cells through MAPK and NF-κB. Am J Pathol. 182:2310–2321. 2013. View Article : Google Scholar : PubMed/NCBI
26	Huang M, Wang HQ, Zhang Q, Yan XD, Hao M and Luo ZJ: Alterations of ADAMTSs and TIMP-3 in human nucleus pulposus cells subjected to compressive load: Implications in the pathogenesis of human intervertebral disc degeneration. J Orthop Res. 30:267–273. 2012. View Article : Google Scholar
27	Smolders LA, Meij BP, Onis D, Riemers FM, Bergknut N, Wubbolts R, Grinwis GC, Houweling M, Groot Koerkamp MJ, van Leenen D, et al: Gene expression profiling of early intervertebral disc degeneration reveals a down-regulation of canonical Wnt signaling and caveolin-1 expression: implications for development of regenerative strategies. Arthritis Res Ther. 15:R232013. View Article : Google Scholar : PubMed/NCBI
28	Sanges D, Romo N, Simonte G, Di Vicino U, Tahoces AD, Fernández E and Cosma MP: Wnt/β-catenin signaling triggers neuron reprogramming and regeneration in the mouse retina. Cell Rep. 4:271–286. 2013. View Article : Google Scholar : PubMed/NCBI
29	Iwata M, Aikawa T, Hakozaki T, Arai K, Ochi H, Haro H, Tagawa M, Asou Y and Hara Y: Enhancement of Runx2 expression is potentially linked to β-catenin accumulation in canine intervertebral disc degeneration. J Cell Physiol. 230:180–190. 2015. View Article : Google Scholar