Downregulation of aquaporin 9 decreases catabolic factor expression through nuclear factor‑κB signaling in chondrocytes

Takeuchi,Kazuhiro; Hayashi,Shinya; Matumoto,Tomoyuki; Hashimoto,Shingo; Takayama,Koji; Chinzei,Nobuaki; Kihara,Shinsuke; Haneda,Masahiko; Kirizuki,Shinsuke; Kuroda,Yuichi; Tsubosaka,Masanori; Nishida,Kotaro; Kuroda,Ryosuke

doi:10.3892/ijmm.2018.3729

Downregulation of aquaporin 9 decreases catabolic factor expression through nuclear factor‑κB signaling in chondrocytes

Authors:
- Kazuhiro Takeuchi
- Shinya Hayashi
- Tomoyuki Matumoto
- Shingo Hashimoto
- Koji Takayama
- Nobuaki Chinzei
- Shinsuke Kihara
- Masahiko Haneda
- Shinsuke Kirizuki
- Yuichi Kuroda
- Masanori Tsubosaka
- Kotaro Nishida
- Ryosuke Kuroda
View Affiliations

Affiliations: Department of Orthopedic Surgery, Kobe University Graduate School of Medicine, Kobe 650‑0017, Japan
Published online on: June 13, 2018 https://doi.org/10.3892/ijmm.2018.3729
Pages: 1548-1558

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Aquaporins (AQPs) are small integral membrane proteins that are essential for water transport across membranes. AQP9, one of the 13 mammalian AQPs (including AQP0 to 12), has been reported to be highly expressed in hydrarthrosis and synovitis patients. Given that several studies have identified signal transduction as an additional function of AQPs, it is hypothesized that AQP9 may modulate inflammatory signal transduction in chondrocytes. Therefore, the present study used a model of interleukin (IL)‑1β‑induced inflammation to determine the mechanisms associated with AQP9 functions in chondrocytes. Osteoarthritis (OA) and normal cartilage samples were subjected to immunohistological analysis. In addition, matrix metalloproteinase (MMP)3, MMP13 and a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS‑5) mRNA and protein analysis was conducted in normal human articular chondrocytes from the knee (NHAC‑Kn) stimulated with IL‑1β by reverse transcription‑polymerase chain reaction (RT‑qPCR) and western blotting, respectively. AQP9 knockdown was also performed by transfection of AQP9‑specific small interfering RNA using Lipofectamine. AQP1, 3, 7, 9 and 11 mRNA expression levels were detected in OA human chondrocytes and in IL‑1β‑treated normal human chondrocytes. The levels of AQP9, MMP‑3, MMP‑13 and ADAMTS‑5 mRNA were increased by treatment with 10 ng/ml IL‑1β in a time‑dependent manner, while knockdown of AQP9 expression significantly decreased the mRNA levels of the MMP3, MMP13 and ADAMTS‑5 genes, as well as the phosphorylation of IκB kinase (IKK). Treatment with a specific IKK inhibitor also significantly decreased the expression levels of MMP‑3, MMP‑13 and ADAMTS‑5 in response to IL‑1β stimulation. Furthermore, immunohistochemical analysis demonstrated that AQP9 and inflammatory markers were highly expressed in OA cartilage. In addition, the downregulation of AQP9 in cultured chondrocytes decreased the catabolic gene expression in response to IL‑1β stimulation through nuclear factor‑κB signaling. Therefore, AQP9 may be a promising target for the treatment of OA.

View Figures

View References

1	Chevalier X, Groult N, Emod I and Planchenault T: Proteoglycan-degrading activity associated with the 40 kDa collagen-binding fragment of fibronectin. Br J Rheumatol. 35:506–514. 1996. View Article : Google Scholar : PubMed/NCBI
2	Berenbaum F and van den Berg WB: Inflammation in osteoarthritis: Changing views. Osteoarthritis Cartilage. 23:1823–1824. 2015. View Article : Google Scholar : PubMed/NCBI
3	Berenbaum F: Osteoarthritis as an inflammatory disease (osteoarthritis is not osteoarthrosis!). Osteoarthritis Cartilage. 21:16–21. 2013. View Article : Google Scholar
4	Fernandes JC, Martel-Pelletier J and Pelletier JP: The role of cytokines in osteoarthritis pathophysiology. Biorheology. 39:237–246. 2002.PubMed/NCBI
5	Goldring MB: The role of cytokines as inflammatory mediators in osteoarthritis: Lessons from animal models. Connect Tissue Res. 40:1–11. 1999. View Article : Google Scholar
6	Loeser RF: Molecular mechanisms of cartilage destruction: Mechanics, inflammatory mediators, and aging collide. Arthritis Rheum. 54:1357–1360. 2006. View Article : Google Scholar : PubMed/NCBI
7	Wojdasiewicz P, Poniatowski LA and Szukiewicz D: The role of inflammatory and anti-inflammatory cytokines in the pathogenesis of osteoarthritis. Mediators Inflamm. 2014:5614592014. View Article : Google Scholar : PubMed/NCBI
8	Shakibaei M, Schulze-Tanzil G, John T and Mobasheri A: Curcumin protects human chondrocytes from IL-l1beta-induced inhibition of collagen type II and beta1-integrin expression and activation of caspase-3: An immunomorphological study. Ann Anat. 187:487–497. 2005. View Article : Google Scholar : PubMed/NCBI
9	Stove J, Huch K, Gunther KP and Scharf HP: Interleukin-1beta induces different gene expression of stromelysin, aggrecan and tumor-necrosis-factor-stimulated gene 6 in human osteoarthritic chondrocytes in vitro. Pathobiology. 68:144–149. 2000. View Article : Google Scholar
10	Mengshol JA, Vincenti MP, Coon CI, Barchowsky A and Brinckerhoff CE: Interleukin-1 induction of collagenase 3 (matrix metalloproteinase 13) gene expression in chondrocytes requires p38, c-Jun N-terminal kinase, and nuclear factor kappaB: Differential regulation of collagenase 1 and collagenase 3. Arthritis Rheum. 43:801–811. 2000. View Article : Google Scholar : PubMed/NCBI
11	Matsushita T, Sasaki H, Takayama K, Ishida K, Matsumoto T, Kubo S, Matsuzaki T, Nishida K, Kurosaka M and Kuroda R: The overexpression of SIRT1 inhibited osteoarthritic gene expression changes induced by interleukin-1β in human chondrocytes. J Orthop Res. 31:531–537. 2013. View Article : Google Scholar
12	Wang M, Sampson ER, Jin H, Li J, Ke QH, Im HJ and Chen D: MMP13 is a critical target gene during the progression of osteoarthritis. Arthritis Res Ther. 15:R52013. View Article : Google Scholar : PubMed/NCBI
13	Miller RE, Tran PB, Ishihara S, Larkin J and Malfait AM: Therapeutic effects of an anti-ADAMTS-5 antibody on joint damage and mechanical allodynia in a murine model of osteoarthritis. Osteoarthritis Cartilage. 24:299–306. 2016. View Article : Google Scholar :
14	Ishibashi K, Hara S and Kondo S: Aquaporin water channels in mammals. Clin Exp Nephrol. 13:107–117. 2009. View Article : Google Scholar
15	Tsukaguchi H, Weremowicz S, Morton CC and Hediger MA: Functional and molecular characterization of the human neutral solute channel aquaporin-9. Am J Physiol. 277:F685–F696. 1999.PubMed/NCBI
16	Ishibashi K: New members of mammalian aquaporins: AQP10-AQP12. Handb Exp Pharmacol. 251–262. 2009. View Article : Google Scholar
17	Verkman AS: Roles of aquaporins in kidney revealed by transgenic mice. Semin Nephrol. 26:200–208. 2006. View Article : Google Scholar : PubMed/NCBI
18	Sohara E, Rai T, Sasaki S and Uchida S: Physiological roles of AQP7 in the kidney: Lessons from AQP7 knockout mice. Biochim Biophys Acta. 1758:1106–1110. 2006. View Article : Google Scholar : PubMed/NCBI
19	Ko SB, Mizuno N, Yatabe Y, Yoshikawa T, Ishiguro H, Yamamoto A, Azuma S, Naruse S, Yamao K, Muallem S and Goto H: Aquaporin 1 water channel is overexpressed in the plasma membranes of pancreatic ducts in patients with autoimmune pancreatitis. J Med Invest. 56(Suppl): 318–321. 2009. View Article : Google Scholar : PubMed/NCBI
20	Huysseune S, Kienlen-Campard P, Hebert S, Tasiaux B, Leroy K, Devuyst O, Brion JP, De Strooper B and Octave JN: Epigenetic control of aquaporin 1 expression by the amyloid precursor protein. FASEB J. 23:4158–4167. 2009. View Article : Google Scholar : PubMed/NCBI
21	Mobasheri A, Moskaluk CA, Marples D and Shakibaei M: Expression of aquaporin 1 (AQP1) in human synovitis. Ann Anat. 192:116–121. 2010. View Article : Google Scholar : PubMed/NCBI
22	Rojek AM, Skowronski MT, Fuchtbauer EM, Füchtbauer AC, Fenton RA, Agre P, Frøkiaer J and Nielsen S: Defective glycerol metabolism in aquaporin 9 (AQP9) knockout mice. Proc Natl Acad Sci USA. 104:3609–3614. 2007. View Article : Google Scholar : PubMed/NCBI
23	Spegel P, Chawade A, Nielsen S, Kjellbom P and Rutzler M: Deletion of glycerol channel aquaporin-9 (Aqp9) impairs long-term blood glucose control in C57BL/6 leptin receptor-deficient (db/db) obese mice. Physiol Rep. 3:e125382015. View Article : Google Scholar : PubMed/NCBI
24	Naka M, Kanamori A, Negi A and Nakamura M: Reduced expression of aquaporin-9 in rat optic nerve head and retina following elevated intraocular pressure. Invest Ophthalmol Vis Sci. 51:4618–4626. 2010. View Article : Google Scholar : PubMed/NCBI
25	de Baey A and Lanzavecchia A: The role of aquaporins in dendritic cell macropinocytosis. J Exp Med. 191:743–748. 2000. View Article : Google Scholar : PubMed/NCBI
26	Zhu N, Feng X, He C, Gao H, Yang L, Ma Q, Guo L, Qiao Y, Yang H and Ma T: Defective macrophage function in aquaporin-3 deficiency. FASEB J. 25:4233–4239. 2011. View Article : Google Scholar : PubMed/NCBI
27	Rabolli V, Wallemme L, Lo Re S, Uwambayinema F, Palmai-Pallag M, Thomassen L, Tyteca D, Octave JN, Marbaix E, Lison D, et al: Critical role of aquaporins in interleukin 1β(IL-1β)-induced inflammation. J Biol Chem. 289:13937–13947. 2014. View Article : Google Scholar : PubMed/NCBI
28	Nagahara M, Waguri-Nagaya Y, Yamagami T, Aoyama M, Tada T, Inoue K, Asai K and Otsuka T: TNF-alpha-induced aquaporin 9 in synoviocytes from patients with OA and RA. Rheumatology (Oxford). 49:898–906. 2010. View Article : Google Scholar
29	General Assembly of the World Medical Association: World medical association declaration of helsinki: Ethical principles for medical research involving human subjects. J Am Coll Dent. 81:14–18. 2014.
30	Mankin HJ, Dorfman H, Lippiello L and Zarins A: Biochemical and metabolic abnormalities in articular cartilage from osteo-arthritic human hips. II Correlation of morphology with biochemical and metabolic data. J Bone Joint Surg Am. 53:523–537. 1971. View Article : Google Scholar : PubMed/NCBI
31	Pattoli MA, MacMaster JF, Gregor KR and Burke JR: Collagen and aggrecan degradation is blocked in interleukin-1-treated cartilage explants by an inhibitor of IkappaB kinase through suppression of metalloproteinase expression. J Pharmacol Exp Ther. 315:382–388. 2005. View Article : Google Scholar : PubMed/NCBI
32	Kihara S, Hayashi S, Hashimoto S, Kanzaki N, Takayama K, Matsumoto T, Chinzei N, Iwasa K, Haneda M, Takeuchi K, et al: Cyclin-dependent kinase inhibitor-1-deficient mice are susceptible to osteoarthritis associated with enhanced inflammation. J Bone Miner Res. 32:991–1001. 2017. View Article : Google Scholar : PubMed/NCBI
33	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
34	Matsushima A, Ogura H, Koh T, Shimazu T and Sugimoto H: Enhanced expression of aquaporin 9 in activated polymor-phonuclear leukocytes in patients with systemic inflammatory response syndrome. Shock. 42:322–326. 2014. View Article : Google Scholar : PubMed/NCBI
35	Mesko B, Poliska S, Szegedi A, Szekanecz Z, Palatka K, Papp M and Nagy L: Peripheral blood gene expression patterns discriminate among chronic inflammatory diseases and healthy controls and identify novel targets. BMC Med Genomics. 3:152010. View Article : Google Scholar : PubMed/NCBI
36	Gilmore TD: The Rel/NF-kappaB signal transduction pathway: Introduction. Oncogene. 18:6842–6844. 1999. View Article : Google Scholar : PubMed/NCBI
37	Holgate ST: Cytokine and anti-cytokine therapy for the treatment of asthma and allergic disease. Cytokine. 28:152–157. 2004. View Article : Google Scholar : PubMed/NCBI
38	Williams RO, Paleolog E and Feldmann M: Cytokine inhibitors in rheumatoid arthritis and other autoimmune diseases. Curr Opin Pharmacol. 7:412–417. 2007. View Article : Google Scholar : PubMed/NCBI
39	Liu SF and Malik AB: NF-kappa B activation as a pathological mechanism of septic shock and inflammation. Am J Physiol Lung Cell Mol Physiol. 290:L622–L645. 2006. View Article : Google Scholar : PubMed/NCBI
40	Wang BF, Cui ZW, Zhong ZH, Sun YH, Sun QF, Yang GY and Bian LG: Curcumin attenuates brain edema in mice with intracerebral hemorrhage through inhibition of AQP4 and AQP9 expression. Acta Pharmacol Sin. 36:939–948. 2015. View Article : Google Scholar : PubMed/NCBI
41	Yang M, Gao F, Liu H, Yu WH and Sun SQ: Temporal changes in expression of aquaporin-3, -4, -5 and -8 in rat brains after permanent focal cerebral ischemia. Brain Res. 1290:121–132. 2009. View Article : Google Scholar : PubMed/NCBI
42	Song TT, Bi YH, Gao YQ, Huang R, Hao K, Xu G, Tang JW, Ma ZQ, Kong FP, Coote JH, et al: Systemic pro-inflammatory response facilitates the development of cerebral edema during short hypoxia. J Neuroinflammation. 13:632016. View Article : Google Scholar : PubMed/NCBI