Carboxymethyl‑chitosan attenuates inducible nitric oxide synthase and promotes interleukin‑10 production in rat chondrocytes

Kong,Ying; Zhang,Yuanmin; Zhao,Xiaowei; Wang,Guodong; Liu,Qingkuan

doi:10.3892/etm.2017.5258

Carboxymethyl‑chitosan attenuates inducible nitric oxide synthase and promotes interleukin‑10 production in rat chondrocytes

Authors:
- Ying Kong
- Yuanmin Zhang
- Xiaowei Zhao
- Guodong Wang
- Qingkuan Liu
View Affiliations

Affiliations: Department of Bone and Joint Surgery, Affiliated Hospital of Jining Medical University, Jining, Shandong 272029, P.R. China
Published online on: October 3, 2017 https://doi.org/10.3892/etm.2017.5258
Pages: 5641-5646

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

Osteoarthritis (OA) is a common age‑related degenerative joint disease, which is caused by the breakdown of joint cartilage and the underlying bone. Carboxymethyl (CM)‑chitosan is a soluble derivative of chitosan that has similar physicochemical properties to the extracellular proteoglycans identified in hyaline cartilage. Previous studies have demonstrated that CM‑chitosan serves a protective role in a rabbit OA model. The aim of the present study was to investigate the effect of CM‑chitosan on NO production and inflammation through its upregulation of interleukin (IL)‑10, and the activation of the janus kinase (JAK)/signal transducer and activator of transcription (STAT)/suppressor of cytokine signaling (SOCS) signaling pathway. In the present study primary rat chondrocytes were induced to inflammation with 2 µg/ml lipopolysaccharide. The cells were subsequently subjected to increasing concentrations of CM‑chitosan (50, 100 and 200 µg/ml) and the relative mRNA and protein expression of inducible nitric oxide synthase (iNOS), IL‑10, JAK1, STAT3 and SOCS3 were measured by RT‑qPCR and western blot analysis respectively. The results revealed that CM‑chitosan attenuated inflammation by significantly reducing iNOS expression and upregulating the anti‑inflammatory cytokine IL‑10 in a dose‑dependent manner (P<0.05). The expression of JAK1, STAT3 and SOCS3 were also significantly upregulated by CM‑chitosan (all P<0.05). The protective role of CM‑chitosan against NO production was due to its upregulation of IL‑10 and its activation of the JAK/STAT/SOCS signaling pathway.

View Figures

View References

1	Kidd BL: Osteoarthritis and joint pain. Pain. 123:6–9. 2006. View Article : Google Scholar : PubMed/NCBI
2	Rogers MW and Wilder FV: The association of BMI and knee pain among persons with radiographic knee osteoarthritis: A cross-sectional study. BMC Musculoskelet Disord. 9:1632008. View Article : Google Scholar : PubMed/NCBI
3	Zhang Y and Jordan JM: Epidemiology of osteoarthritis. Clin Geriatr Med. 26:355–369. 2010. View Article : Google Scholar : PubMed/NCBI
4	Le LT, Swingler TE and Clark IM: Review: The role of microRNAs in osteoarthritis and chondrogenesis. Arthritis Rheum. 65:1963–1974. 2013. View Article : Google Scholar : PubMed/NCBI
5	Wheaton AJ, Borthakur A, Shapiro EM, Regatte RR, Akella SV, Kneeland JB and Reddy R: Proteoglycan loss in human knee cartilage: Quantitation with sodium MR imaging-seasibility study. Radiology. 231:900–905. 2004. View Article : Google Scholar : PubMed/NCBI
6	Clements KM, Price JS, Chambers MG, Visco DM, Poole AR and Mason RM: Gene deletion of either interleukin-1beta, interleukin-1beta-converting enzyme, inducible nitric oxide synthase, or stromelysin 1 accelerates the development of knee osteoarthritis in mice after surgical transection of the medial collateral ligament and partial medial meniscectomy. Arthritis Rheum. 48:3452–3463. 2003. View Article : Google Scholar : PubMed/NCBI
7	Murrell GA, Jang D and Williams RJ: Nitric oxide activates metalloprotease enzymes in articular cartilage. Biochem Biophys Res Commun. 206:15–21. 1995. View Article : Google Scholar : PubMed/NCBI
8	Hashimoto S, Takahashi K, Amiel D, Coutts RD and Lotz M: Chondrocyte apoptosis and nitric oxide production during experimentally induced osteoarthritis. Arthritis Rheum. 41:1266–1274. 1998. View Article : Google Scholar : PubMed/NCBI
9	Farrell AJ, Blake DR, Palmer RM and Moncada S: Increased concentrations of nitrite in synovial fluid and serum samples suggest increased nitric oxide synthesis in rheumatic diseases. Ann Rheum Dis. 51:1219–1222. 1992. View Article : Google Scholar : PubMed/NCBI
10	Vuolteenaho K, Moilanen T, Jalonen U, Lahti A, Nieminen R, van Beuningen HM, van der Kraan PM and Moilanen E: TGFbeta inhibits IL-1 -induced iNOS expression and NO production in immortalized chondrocytes. Inflamm Res. 54:420–427. 2005. View Article : Google Scholar : PubMed/NCBI
11	Gómez R, Scotece M, Conde J, Lopez V, Pino J, Lago F, Gómez-Reino JJ and Gualillo O: Nitric oxide boosts TLR-4 mediated lipocalin 2 expression in chondrocytes. J Orthop Res. 31:1046–1052. 2013. View Article : Google Scholar : PubMed/NCBI
12	Takada K, Hirose J, Yamabe S, Uehara Y and Mizuta H: Endoplasmic reticulum stress mediates nitric oxide-induced chondrocyte apoptosis. Biomed Rep. 1:315–319. 2013. View Article : Google Scholar : PubMed/NCBI
13	de Girolamo L, Kon E, Filardo G, Marmotti AG, Soler F, Peretti GM, Vannini F, Madry H and Chubinskaya S: Regenerative approaches for the treatment of early OA. Knee Surg Sports Traumatol Arthrosc. 24:1826–1835. 2016. View Article : Google Scholar : PubMed/NCBI
14	Lahiji A, Sohrabi A, Hungerford DS and Frondoza CG: Chitosan supports the expression of extracellular matrix proteins in human osteoblasts and chondrocytes. J Biomed Mater Res. 51:586–595. 2000. View Article : Google Scholar : PubMed/NCBI
15	Oprenyeszk F, Sanchez C, Dubuc JE, Maquet V, Henrist C, Compère P and Henrotin Y: Chitosan enriched three-dimensional matrix reduces inflammatory and catabolic mediators production by human chondrocytes. PLoS One. 10:e01283622015. View Article : Google Scholar : PubMed/NCBI
16	Xia W, Liu P, Zhang J and Chen J: Biological activities of chitosan and chitooligosaccharides. Food Hydrocolloid. 25:170–179. 2011. View Article : Google Scholar
17	Jung WJ and Park RD: Bioproduction of chitooligosaccharides: Present and perspectives. Mar Drugs. 12:5328–5356. 2014. View Article : Google Scholar : PubMed/NCBI
18	Kerch G: The potential of chitosan and its derivatives in prevention and treatment of age-related diseases. Mar Drugs. 13:2158–2182. 2015. View Article : Google Scholar : PubMed/NCBI
19	Croisier F and Jérôme C: Chitosan-based biomaterials for tissue engineering. Eur Polym J. 49:780–792. 2013. View Article : Google Scholar
20	Chen Q, Liu SQ, Du YM, Peng H and Sun LP: Carboxymethyl-chitosan protects rabbit chondrocytes from interleukin-1beta-induced apoptosis. Eur J Pharmacol. 541:1–8. 2006. View Article : Google Scholar : PubMed/NCBI
21	Liu SQ, Qiu B, Chen LY, Peng H and Du YM: The effects of carboxymethylated chitosan on metalloproteinase-1, −3 and tissue inhibitor of metalloproteinase-1 gene expression in cartilage of experimental osteoarthritis. Rheumatol Int. 26:52–57. 2005. View Article : Google Scholar : PubMed/NCBI
22	Yu H, Liu Z, Zhou H, Dai W, Chen S, Shu Y and Feng J: JAK-STAT pathway modulates the roles of iNOS and COX-2 in the cytoprotection of early phase of hydrogen peroxide preconditioning against apoptosis induced by oxidative stress. Neurosci Lett. 529:166–171. 2012. View Article : Google Scholar : PubMed/NCBI
23	Dell'Albani P, Santangelo R, Torrisi L, Nicoletti VG, De Vellis J and Giuffrida Stella AM: JAK/STAT signaling pathway mediates cytokine-induced iNOS expression in primary astroglial cell cultures. J Neurosci Res. 65:417–424. 2001. View Article : Google Scholar : PubMed/NCBI
24	Séguin CA and Bernier SM: TNFalpha suppresses link protein and type II collagen expression in chondrocytes: Role of MEK1/2 and NF-kappaB signaling pathways. J Cell Physiol. 197:356–369. 2003. View Article : Google Scholar : PubMed/NCBI
25	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 : PubMed/NCBI
26	Goff WL, O'Rourke KI, Johnson WC, Lacy PA, Davis WC and Wyatt CR: The role of IL-10 in iNOS and cytokine mRNA expression during in vitro differentiation of bovine mononuclear phagocytes. J Interf Cytok Res. 18:139–149. 1998. View Article : Google Scholar
27	Bijlsma JW, Berenbaum F and Lafeber FP: Osteoarthritis: An update with relevance for clinical practice. Lancet. 377:2115–2126. 2011. View Article : Google Scholar : PubMed/NCBI
28	Zheng S, Hunter DJ, Xu J and Ding C: Monoclonal antibodies for the treatment of osteoarthritis. Expert Opin Biol Ther. 16:1529–1540. 2016. View Article : Google Scholar : PubMed/NCBI
29	Ma Z, Wang Y, Piao T and Liu J: Echinocystic acid inhibits IL-1β-induced COX-2 and iNOS expression in human osteoarthritis chondrocytes. Inflammation. 39:543–549. 2016. View Article : Google Scholar : PubMed/NCBI
30	Cheng AW, Stabler TV, Bolognesi MP and Kraus VB: Selenomethionine inhibits IL-1β inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX2) expression in primary human chondrocytes. Osteoarthritis Cartilage. 19:118–125. 2011. View Article : Google Scholar : PubMed/NCBI
31	Ying X, Chen X, Cheng S, Shen Y, Peng L and Xu HZ: Piperine inhibits IL-β induced expression of inflammatory mediators in human osteoarthritis chondrocyte. Int Immunopharmaco. 17:293–299. 2013. View Article : Google Scholar
32	Azuma K, Osaki T, Minami S and Okamoto Y: Anticancer and anti-inflammatory properties of chitin and chitosan oligosaccharides. J Funct Biomater. 6:33–49. 2015. View Article : Google Scholar : PubMed/NCBI
33	Martel-Pelletier J: Pathophysiology of osteoarthritis. Osteoarthritis Cartilage. 12 Suppl A:S31–S33. 2004. View Article : Google Scholar : PubMed/NCBI
34	Namba A, Aida Y, Suzuki N, Watanabe Y, Kawato T, Motohashi M, Maeno M, Matsumura H and Matsumoto M: Effects of IL-6 and soluble IL-6 receptor on the expression of cartilage matrix proteins in human chondrocytes. Connect Tissue Res. 48:263–270. 2007. View Article : Google Scholar : PubMed/NCBI
35	Lin PM, Chen CC and Torzilli PA: Increased stromelysin-1 (MMP-3), proteoglycan degradation (3B3- and 7D4) and collagen damage in cyclically load-injured articular cartilage. Osteoarthritis Cartilage. 12:485–496. 2004. View Article : Google Scholar : PubMed/NCBI
36	Porée B, Kypriotou M, Chadjichristos C, Beauchef G, Renard E, Legendre F, Melin M, Gueret S, Hartmann DJ, Malléin-Gerin F, et al: Interleukin-6 (IL-6) and/or soluble IL-6 receptor down-regulation of human type II collagen gene expression in articular chondrocytes requires a decrease of Sp1.Sp3 ratio and of the binding activity of both factors to the COL2A1 promoter. J Biol Chem. 283:4850–4865. 2008. View Article : Google Scholar : PubMed/NCBI
37	Imamura M, Targino RA, Hsing WT, Imamura S, Azevedo RS, Boas LS, Tozetto-Mendoza TR, Alfieri FM, Filippo TR and Battistella LR: Concentration of cytokines in patients with osteoarthritis of the knee and fibromyalgia. Clin Interv Aging. 9:939–944. 2014.PubMed/NCBI
38	Rojas-Ortega M, Cruz R, Vega-López MA, Cabrera-González M, Hernández-Hernández JM, Lavalle-Montalvo C and Kouri JB: Exercise modulates the expression of IL-1β and IL-10 in the articular cartilage of normal and osteoarthritis-induced rats. Pathol Res Pract. 211:435–443. 2015. View Article : Google Scholar : PubMed/NCBI
39	Li S, Wan J, Anderson W, Sun H, Zhang H, Peng X, Yu Z, Wang T, Yan X and Smith W: Downregulation of IL-10 secretion by Treg cells in osteoarthritis is associated with a reduction in Tim-3 expression. Biomed Pharmacothr. 79:159–165. 2016. View Article : Google Scholar
40	Müller RD, John T, Kohl B, Oberholzer A, Gust T, Hostmann A, Hellmuth M, Laface D, Hutchins B, Laube G, et al: IL-10 overexpression differentially affects cartilage matrix gene expression in response to TNF-alpha in human articular chondrocytes in vitro. Cytokine. 44:377–385. 2008. View Article : Google Scholar : PubMed/NCBI
41	Josephson K, Logsdon NJ and Walter MR: Crystal structure of the IL-10/IL-10R1 complex reveals a shared receptor binding site. Immunity. 15:35–46. 2001. View Article : Google Scholar : PubMed/NCBI
42	Carey AJ, Tan CK and Ulett GC: Infection-induced IL-10 and JAK-STAT: A review of the molecular circuitry controlling immune hyperactivity in response to pathogenic microbes. JAKSTAT. 1:159–167. 2012.PubMed/NCBI