Angiopoietin-like 2 upregulation promotes human chondrocyte injury via NF-κB and p38/MAPK signaling pathway
- 147 Downloads
Several cellular and molecular processes participate in the pathologic changes of osteoarthritis (OA). However, the core molecular regulators of these processes are unclear, and no effective treatment for OA disease has been developed so far. ANGPTL2 is well known for its tissue remolding and pro-inflammation properties. However, the role of ANGPTL2 in osteoarthritis (OA) still remains unclear. To explore the expression level of ANGPTL2 in human OA cartilage and investigate the function of ANGPTL2 in human chondrocytes injury, qRT-PCR, western blot and immunohistochemistry were employed to investigate the expression of ANGPTL2 between human OA and normal cartilage samples. Next, human primary chondrocytes were treated with IL-1β to mimic OA progress in vitro, and the expression of ANGPTL2 were tested by qRT-PCR and western blot. Furthermore, the effect of ANGPTL2 in the expression of pro-inflammation cytokines (IL-1β, IL-6), proteolytic enzymes (MMP-1, MMP-13) and component of the cartilage matrix (COL2A1 and aggrecan) in human primary chondrocyte were explored by gain-of-function and loss-of-function methods. Finally, the nuclear factor kappa B (NF-κB) and p38/MAPK signaling pathways were also tested by western blot analysis. In this study, firstly, the expression level of ANGPTL2 was elevated both in human OA cartilage samples and IL-1β stimulated human chondrocytes. Secondly, ANGPTL2 upregulation promotes extracellular matrix (ECM) degradation and inflammation mediator production in human chondrocytes. Finally, ANGPTL2 activated the NF-κB and p38/MAPK signaling pathways via integrin α5β1. This study, for the first time, highlights that ANGPTL2 secreted by human chondrocytes plays a negative role in the pathogenesis of osteoarthritis, and it may be a potential therapeutic target in OA.
KeywordsOsteoarthritis ANGPTL2 ECM degradation Inflammation mediators production
Nuclear factor kB
Mitogen-activated protein kinase
We thank Kai Lin, Yinglei Fang, Penghui Ke for assistance in collecting cartilage samples.
Zongsheng Yin, Wei He and Jiegou Xu designed the study. Wenshan Shan, Zhenfei Ding, Guanjun Cui performed in vitro experiments. Chao Chen, Wei Huang, Wei Lu and Fuen Liu detected the expression level of ANGPTL2 in OA cartilage samples. Sha Luo participated in the supplementary experiment. Wenshan Shan wrote the paper.
This work was supported by grants from the National Natural Science Foundation of China (No. 81672161) and National Undergraduate Traning Programs for Innovation and Entrepreneurship-China (No. 201810366025).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no competing interests.
All cartilage samples were obtained from the First Affiliated Hospital of Anhui Medical University. This research was approved by the Ethics Committee of the First Affiliated Hospital of Anhui Medical University.
All patients obtained a written informed consent about the study.
- 3.Echtermeyer F, Bertrand J, Dreier R, Meinecke I, Neugebauer K, Fuerst M, Lee YJ, Song YW, Herzog C, Theilmeier G, Pap T (2009) Syndecan-4 regulates ADAMTS-5 activation and cartilage breakdown in osteoarthritis (in eng). Nat Med 15:1072–1076. https://doi.org/10.1038/nm.1998 CrossRefPubMedGoogle Scholar
- 5.Neuhold LA, Killar L, Zhao W, Sung ML, Warner L, Kulik J, Turner J, Wu W, Billinghurst C, Meijers T, Poole AR, Babij P, DeGennaro LJ (2001) Postnatal expression in hyaline cartilage of constitutively active human collagenase-3 (MMP-13) induces osteoarthritis in mice (in eng). J Clin Investig 107:35–44. https://doi.org/10.1172/jci10564 CrossRefPubMedGoogle Scholar
- 7.Chen WP, Hu ZN, Jin LB, Wu LD (2017) Licochalcone A Inhibits MMPs and ADAMTSs via the NF-kappaB and Wnt/beta-Catenin Signaling Pathways in Rat Chondrocytes (in eng). Cellular physiology and biochemistry: international journal of experimental cellular physiology, biochemistry, and pharmacology 43:937–944. https://doi.org/10.1159/000481645 CrossRefGoogle Scholar
- 8.Yuan Y, Tan H, Dai P (2017) Kruppel-Like Factor 2 Regulates Degradation of Type II Collagen by Suppressing the Expression of Matrix Metalloproteinase (MMP)-13 (in eng). Cellular physiology and biochemistry: international journal of experimental cellular physiology, biochemistry, and pharmacology 42:2159–2168. https://doi.org/10.1159/000479991 CrossRefGoogle Scholar
- 10.Son YO, Park S, Kwak JS, Won Y, Choi WS, Rhee J, Chun CH, Ryu JH, Kim DK, Choi HS, Chun JS (2017) Estrogen-related receptor gamma causes osteoarthritis by upregulating extracellular matrix-degrading enzymes (in eng). Nature communications 8:2133. https://doi.org/10.1038/s41467-017-01868-8 CrossRefPubMedPubMedCentralGoogle Scholar
- 11.Pelletier JP, Martel-Pelletier J, Abramson SB (2001) Osteoarthritis, an inflammatory disease: potential implication for the selection of new therapeutic targets (in eng). Arthritis Rheum 44:1237–1247. https://doi.org/10.1002/1529-0131(200106)44:6%3c1237:AID-ART214%3e3.0.CO;2-F CrossRefPubMedGoogle Scholar
- 14.Bradley JD, Brandt KD, Katz BP, Kalasinski LA, Ryan SI (1991) Comparison of an antiinflammatory dose of ibuprofen, an analgesic dose of ibuprofen, and acetaminophen in the treatment of patients with osteoarthritis of the knee (in eng). The New England journal of medicine 325:87–91. https://doi.org/10.1056/nejm199107113250203 CrossRefPubMedGoogle Scholar
- 15.da Costa BR, Reichenbach S, Keller N, Nartey L, Wandel S, Juni P, Trelle S (2016) RETRACTED: effectiveness of non-steroidal anti-inflammatory drugs for the treatment of pain in knee and hip osteoarthritis: a network meta-analysis (in eng). Lancet (London, England) 387:2093–2105. https://doi.org/10.1016/s0140-6736(16)30002-2 CrossRefGoogle Scholar
- 22.Aoi J, Endo M, Kadomatsu T, Miyata K, Nakano M, Horiguchi H, Ogata A, Odagiri H, Yano M, Araki K, Jinnin M, Ito T, Hirakawa S, Ihn H, Oike Y (2011) Angiopoietin-like protein 2 is an important facilitator of inflammatory carcinogenesis and metastasis (in eng). Can Res 71:7502–7512. https://doi.org/10.1158/0008-5472.can-11-1758 CrossRefGoogle Scholar
- 23.Odagiri H, Kadomatsu T, Endo M, Masuda T, Morioka MS, Fukuhara S, Miyamoto T, Kobayashi E, Miyata K, Aoi J, Horiguchi H, Nishimura N, Terada K, Yakushiji T, Manabe I, Mochizuki N, Mizuta H, Oike Y (2014) The secreted protein ANGPTL2 promotes metastasis of osteosarcoma cells through integrin alpha5beta1, p38 MAPK, and matrix metalloproteinases (in eng). Science Signaling 7:7. https://doi.org/10.1126/scisignal.2004612 CrossRefGoogle Scholar
- 24.Horio E, Kadomatsu T, Miyata K, Arai Y, Hosokawa K et al (2014) Role of endothelial cell-derived angptl2 in vascular inflammation leading to endothelial dysfunction and atherosclerosis progression (in eng). Arterioscler Thromb Vasc Biol 34:790–800. https://doi.org/10.1161/atvbaha.113.303116 CrossRefPubMedGoogle Scholar
- 26.Corciulo C, Lendhey M, Wilder T, Schoen H, Cornelissen AS, Chang G, Kennedy OD, Cronstein BN (2017) Endogenous adenosine maintains cartilage homeostasis and exogenous adenosine inhibits osteoarthritis progression (in eng). Nature communications 8:15019. https://doi.org/10.1038/ncomms15019 CrossRefPubMedPubMedCentralGoogle Scholar
- 27.Jacques C, Bereziat G, Humbert L, Olivier JL, Corvol MT, Masliah J, Berenbaum F (1997) Posttranscriptional effect of insulin-like growth factor-I on interleukin-1beta-induced type II-secreted phospholipase A2 gene expression in rabbit articular chondrocytes (in eng). J Clin Investig 99:1864–1872. https://doi.org/10.1172/jci119353 CrossRefPubMedGoogle Scholar
- 28.Meng F, Li Z, Zhang Z, Yang Z, Kang Y, Zhao X, Long D, Hu S, Gu M, He S, Wu P, Chang Z, He A, Liao W (2018) MicroRNA-193b-3p regulates chondrogenesis and chondrocyte metabolism by targeting HDAC3 (in eng). Theranostics 8:2862–2883. https://doi.org/10.7150/thno.23547 CrossRefPubMedPubMedCentralGoogle Scholar
- 29.Hirasawa M, Takubo K, Osada H, Miyake S, Toda E, Endo M, Umezawa K, Tsubota K, Oike Y, Ozawa Y (2016) Angiopoietin-like Protein 2 Is a Multistep Regulator of Inflammatory Neovascularization in a Murine Model of Age-related Macular Degeneration (in eng). The Journal of biological chemistry 291:7373–7385. https://doi.org/10.1074/jbc.M115.710186 CrossRefPubMedPubMedCentralGoogle Scholar
- 30.Tanoue H, Morinaga J, Yoshizawa T, Yugami M, Itoh H et al (2018) Angiopoietin-like protein 2 promotes chondrogenic differentiation during bone growth as a cartilage matrix factor (in eng). Osteoarthritis and cartilage 26:108–117. https://doi.org/10.1016/j.joca.2017.10.011 CrossRefPubMedGoogle Scholar
- 31.Bose S, Li B, He J, Lv H, Liu Y, Lv X, Zhang C, Zhu Y, Ai D (2019) c-Abl regulates YAPY357 phosphorylation to activate endothelial atherogenic responses to disturbed flow (in eng). Nature communications 129:1167–1179. https://doi.org/10.1038/s41467-019-09453-x10.1172/jci122440 CrossRefGoogle Scholar
- 32.Lee SJ, Lee CK, Kang S, Park I, Kim YH, Kim SK, Hong SP, Bae H, He Y, Kubota Y, Koh GY (2018) Angiopoietin-2 exacerbates cardiac hypoxia and inflammation after myocardial infarction (in eng). J Clin Investig 128:5018–5033. https://doi.org/10.1016/j.ccell.2018.11.01610.1172/jci99659 CrossRefPubMedGoogle Scholar
- 33.Duchet BJ, Hansel CS, Maynard SA, Chow LW, Stevens MM, Sundaram A, Chen C, Khalifeh-Soltani A, Atakilit A, Ren X, Qiu W, Jo H, DeGrado W, Huang X, Sheppard D (2017) Targeting integrin alpha5beta1 ameliorates severe airway hyperresponsiveness in experimental asthma (in eng). ACS Nano 127:365–374. https://doi.org/10.1021/acsnano.6b0597510.1172/jci88555 CrossRefGoogle Scholar
- 36.Karsdal MA, Bay-Jensen AC, Lories RJ, Abramson S, Spector T, Pastoureau P, Christiansen C, Attur M, Henriksen K, Goldring SR, Kraus V (2014) The coupling of bone and cartilage turnover in osteoarthritis: opportunities for bone antiresorptives and anabolics as potential treatments? (in eng). Ann Rheum Dis 73:336–348. https://doi.org/10.1136/annrheumdis-2013-204111 CrossRefPubMedGoogle Scholar
- 39.Billinghurst RC, Dahlberg L, Ionescu M, Reiner A, Bourne R, Rorabeck C, Mitchell P, Hambor J, Diekmann O, Tschesche H, Chen J, Van Wart H, Poole AR (1997) Enhanced cleavage of type II collagen by collagenases in osteoarthritic articular cartilage (in eng). J Clin Investig 99:1534–1545. https://doi.org/10.1172/jci119316 CrossRefPubMedGoogle Scholar
- 40.Lark MW, Bayne EK, Flanagan J, Harper CF, Hoerrner LA, Hutchinson NI, Singer II, Donatelli SA, Weidner JR, Williams HR, Mumford RA, Lohmander LS (1997) Aggrecan degradation in human cartilage. Evidence for both matrix metalloproteinase and aggrecanase activity in normal, osteoarthritic, and rheumatoid joints (in eng). J Clin Investig 100:93–106. https://doi.org/10.1172/jci119526 CrossRefPubMedGoogle Scholar
- 41.de Lange-Brokaar BJ, Ioan-Facsinay A, van Osch GJ, Zuurmond AM, Schoones J, Toes RE, Huizinga TW, Kloppenburg M (2012) Synovial inflammation, immune cells and their cytokines in osteoarthritis: a review (in eng). Osteoarthritis and cartilage 20:1484–1499. https://doi.org/10.1016/j.joca.2012.08.027 CrossRefPubMedGoogle Scholar
- 43.Kobayashi H, Chang SH, Mori D, Itoh S, Hirata M, Hosaka Y, Taniguchi Y, Okada K, Mori Y, Yano F, Chung UI, Akiyama H, Kawaguchi H, Tanaka S, Saito T (2016) Biphasic regulation of chondrocytes by Rela through induction of anti-apoptotic and catabolic target genes (in eng). Nature Communications 7:13336. https://doi.org/10.1038/ncomms13336 CrossRefPubMedPubMedCentralGoogle Scholar
- 46.Pesesse L, Sanchez C, Delcour JP, Bellahcene A, Baudouin C, Msika P, Henrotin Y (2013) Consequences of chondrocyte hypertrophy on osteoarthritic cartilage: potential effect on angiogenesis (in eng). Osteoarthritis and cartilage 21:1913–1923. https://doi.org/10.1016/j.joca.2013.08.018 CrossRefPubMedGoogle Scholar