Icariin possesses chondroprotective efficacy in a rat model of dexamethasone-induced cartilage injury through the activation of miR-206 targeting of cathepsin K

Liu,Ning; Zhang,Tao; Cao,Bo-Ran; Luan,Fei-Yu; Liu,Rui-Xuan; Yin,Hao-Rong; Wang,Wen-Bo

doi:10.3892/ijmm.2017.3289

Icariin possesses chondroprotective efficacy in a rat model of dexamethasone-induced cartilage injury through the activation of miR-206 targeting of cathepsin K

Authors:
- Ning Liu
- Tao Zhang
- Bo-Ran Cao
- Fei-Yu Luan
- Rui-Xuan Liu
- Hao-Rong Yin
- Wen-Bo Wang
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Affiliations: Department of Orthopedic Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China
Published online on: November 27, 2017 https://doi.org/10.3892/ijmm.2017.3289
Pages: 1039-1047

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Abstract

The present study aimed to investigate the articular cartilage and chondrocytes of dexamethasone (DXM)-induced cartilage injuries in rats in response to treatment with icariin, as well as the implicated molecular mechanism. Effects of icariin on bone metabolism and articular cartilage in experimental rats were investigated. Subsequently, the present study assessed dysregulated microRNA (miRNA) in the articular cartilage of experimental rats, and validated the significant miRNA and their targets. Finally, the effects of icariin on chondrocytes in experimental rats and the implicated molecular mechanism were explored. Quantitative polymerase chain reaction demonstrated that icariin significantly reversed DXM-induced bone degradation and stimulated bone regeneration. In addition, some notable changes in articular cartilage were also observed following continuous administration of icariin to DXM-treated rats, including enhanced cartilage area (revealed by safranin-O staining), substantial decrements in serum concentrations of deoxypyridinoline and C-terminal telopeptide of type II collagen, reduced expression of collagen type I and matrix metalloproteinase-13, as well as elevated expression of transforming growth factor-β. Furthermore, miR-206 was determined to be significantly upregulated in the icariin group compared with the DXM-treated group. A luciferase assay further validated cathepsin K as the target RNA of miR-206. Additionally, icariin (100 µM) facilitated the viability of chondrocytes and reduced apoptotic chondrocytes. More importantly, icariin (100 µM) not only abolished the inhibition effect of DXM on miR-206 expression in chondrocytes, but also eliminated the enhancing effect of DXM on cathepsin K expression. Overall, the present study identified icariin as a novel therapeutic agent in DXM-induced cartilage injury in rats, and revealed that the activation of miR-206 targeting of cathepsin K may be responsible for the chondroprotective efficacy of icariin.

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1	Dang AC and Kuo AC: Cartilage biomechanics and implications for treatment of cartilage injuries. Oper Tech Orthop. 24:288–292. 2014. View Article : Google Scholar
2	Mow VC, Ratcliffe A and Poole AR: Cartilage and diarthrodial joints as paradigms for hierarchical materials and structures. Biomaterials. 13:67–97. 1992. View Article : Google Scholar : PubMed/NCBI
3	Frenkel SR, Clancy RM, Ricci JL, Di Cesare PE, Rediske JJ and Abramson SB: Effects of nitric oxide on chondrocyte migration, adhesion, and cytoskeletal assembly. Arthritis Rheum. 39:1905–1912. 1996. View Article : Google Scholar : PubMed/NCBI
4	Klein TJ, Rizzi SC, Reichert JC, Georgi N, Malda J, Schuurman W, Crawford RW and Hutmacher DW: Strategies for zonal cartilage repair using hydrogels. Macromol Biosci. 9:1049–1058. 2009. View Article : Google Scholar : PubMed/NCBI
5	Furman BD, Strand J, Hembree WC, Ward BD, Guilak F and Olson SA: Joint degeneration following closed intraarticular fracture in the mouse knee: a model of posttraumatic arthritis. J Orthop Res. 25:578–592. 2007. View Article : Google Scholar : PubMed/NCBI
6	Seyedin MS and Matava M: Cartilage repair methods. US Patent. 20070128155 A1. Filed December 7, 2006; issued June 7, 2007. https://www.google.ms/patents/US20070128155.
7	Yu S, Chen K, Li S and Zhang K: In vitro and in vivo studies of the effect of a Chinese herb medicine on osteoclastic bone resorption. Chin J Dent Res. 2:7–11. 1999.PubMed/NCBI
8	Zhang DW, Cheng Y, Wang NL, Zhang JC, Yang MS and Yao XS: Effects of total flavonoids and flavonol glycosides from Epimedium koreanum Nakai on the proliferation and differentiation of primary osteoblasts. Phytomedicine. 15:55–61. 2008. View Article : Google Scholar
9	Xie F, Wu CF, Lai WP, Yang XJ, Cheung PY, Yao XS, Leung PC and Wong MS: The osteoprotective effect of Herba epimedii (HEP) extract in vivo and in vitro. Evid Based Complement Alternat Med. 2:353–361. 2005. View Article : Google Scholar : PubMed/NCBI
10	Qian G, Zhang X, Lu L, Wu X, Li S and Meng J: Regulation of Cbfa1 expression by total flavonoids of Herba epimedii. Endocr J. 53:87–94. 2006. View Article : Google Scholar : PubMed/NCBI
11	Hsieh TP, Sheu SY, Sun JS, Chen MH and Liu MH: Icariin isolated from Epimedium pubescens regulates osteoblasts anabolism through BMP-2, SMAD4, and Cbfa1 expression. Phytomedicine. 17:414–423. 2010. View Article : Google Scholar
12	Zhao J, Ohba S, Komiyama Y, Shinkai M, Chung UI and Nagamune T: Icariin: a potential osteoinductive compound for bone tissue engineering. Tissue Eng Part A. 16:233–243. 2010. View Article : Google Scholar
13	Zhang J, Song J and Shao J: Icariin attenuates glucocorticoid-induced bone deteriorations, hypocalcemia and hypercalciuria in mice. Int J Clin Exp Med. 8:7306–7314. 2015.PubMed/NCBI
14	Xu CQ, Liu BJ, Wu JF, Xu YC, Duan XH, Cao YX and Dong JC: Icariin attenuates LPS-induced acute inflammatory responses: involvement of PI3K/Akt and NF-kappaB signaling pathway. Eur J Pharmacol. 642:146–153. 2010. View Article : Google Scholar : PubMed/NCBI
15	Liu MH, Sun JS, Tsai SW, Sheu SY and Chen MH: Icariin protects murine chondrocytes from lipopolysaccharide-induced inflammatory responses and extracellular matrix degradation. Nutr Res. 30:57–65. 2010. View Article : Google Scholar : PubMed/NCBI
16	Qi WN and Scully SP: Type II collagen modulates the composition of extracellular matrix synthesized by articular chondrocytes. J Orthop Res. 21:282–289. 2003. View Article : Google Scholar : PubMed/NCBI
17	Zhou PH, Liu SQ and Peng H: The effect of hyaluronic acid on IL-1β-induced chondrocyte apoptosis in a rat model of osteoarthritis. J Orthop Res. 26:1643–1648. 2008. View Article : Google Scholar : PubMed/NCBI
18	Matsubara J, Yamada Y, Nakajima TE, Kato K, Hamaguchi T, Shirao K, Shimada Y and Shimoda T: Clinical significance of insulin-like growth factor type 1 receptor and epidermal growth factor receptor in patients with advanced gastric cancer. Oncology. 74:76–83. 2008. View Article : Google Scholar : PubMed/NCBI
19	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−ΔΔC(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar
20	Kim JE, Lee MH, Nam DH, Song HK, Kang YS, Lee JE, Kim HW, Cha JJ, Hyun YY, Han SY, et al: Celastrol, an NF-κB inhibitor, improves insulin resistance and attenuates renal injury in db/db mice. PLoS One. 8:e62068. 2013. View Article : Google Scholar
21	DeLaurier A, Jackson B, Pfeiffer D, Ingham K, Horton MA and Price JS: A comparison of methods for measuring serum and urinary markers of bone metabolism in cats. Res Vet Sci. 77:29–39. 2004. View Article : Google Scholar : PubMed/NCBI
22	Tu Y, Xue H, Francis W, Davies AP, Pallister I, Kanamarlapudi V and Xia Z: Lactoferrin inhibits dexamethasone-induced chondrocyte impairment from osteoarthritic cartilage through up-regulation of extracellular signal-regulated kinase 1/2 and suppression of FASL, FAS, and caspase 3. Biochem Biophys Res Commun. 441:249–255. 2013. View Article : Google Scholar : PubMed/NCBI
23	Zhou RR, Jia SF, Zhou Z, Wang Y, Bucana CD and Kleinerman ES: Adenovirus-E1A gene therapy enhances the in vivo sensitivity of Ewing's sarcoma to VP-16. Cancer Gene Ther. 9:407–413. 2002. View Article : Google Scholar : PubMed/NCBI
24	Park JW, Choi YJ, Suh SI, Baek WK, Suh MH, Jin IN, Min DS, Woo JH, Chang JS, Passaniti A, et al: Bcl-2 overexpression attenuates resveratrol-induced apoptosis in U937 cells by inhibition of caspase-3 activity. Carcinogenesis. 22:1633–1639. 2001. View Article : Google Scholar : PubMed/NCBI
25	Inose H, Ochi H, Kimura A, Fujita K, Xu R, Sato S, Iwasaki M, Sunamura S, Takeuchi Y, Fukumoto S, et al: A microRNA regulatory mechanism of osteoblast differentiation. Proc Natl Acad Sci USA. 106:20794–20799. 2009. View Article : Google Scholar : PubMed/NCBI
26	Li XF, Xu H, Zhao YJ, Tang DZ, Xu GH, Holz J, Wang J, Cheng SD, Shi Q and Wang YJ: Icariin augments bone formation and reverses the phenotypes of osteoprotegerin-deficient mice through the activation of Wnt/β-catenin-BMP signaling. Evid Based Complement Alternat Med. 2013:6523172013. View Article : Google Scholar
27	Hsieh TP, Sheu SY, Sun JS and Chen MH: Icariin inhibits osteoclast differentiation and bone resorption by suppression of MAPKs/NF-κB regulated HIF-1α and PGE(2) synthesis. Phytomedicine. 18:176–185. 2011. View Article : Google Scholar
28	Mehl B, Delling G, Schlindwein I, Heilmann P, Voia C, Ziegler R, Nawroth P and Kasperk C: Do markers of bone metabolism reflect the presence of a high- or low-turnover state of bone metabolism? Med Klin. 97:588–594. 2002. View Article : Google Scholar
29	Terzi T, Terzi M, Tander B, Cantürk F and Onar M: Changes in bone mineral density and bone metabolism markers in premenopausal women with multiple sclerosis and the relationship to clinical variables. J Clin Neurosci. 17:1260–1264. 2010. View Article : Google Scholar : PubMed/NCBI
30	Guo Y, Wu X and Zhang X, Li D, Xiao Y, Fan Y and Zhang X: The in vivo study of the effect of icariin on ECM secretion and gene expression of chondrocytes. Pharmacol Clin Chin Mater Med. 2012.
31	Sun P, Liu Y, Deng X, Yu C, Dai N, Yuan X, Chen L, Yu S, Si W, Wang X, et al: An inhibitor of cathepsin K, icariin suppresses cartilage and bone degradation in mice of collagen-induced arthritis. Phytomedicine. 20:975–979. 2013. View Article : Google Scholar : PubMed/NCBI
32	Yuan T, He L, Yang J, Zhang L, Xiao Y, Fan Y and Zhang X: Conjugated icariin promotes tissue-engineered cartilage formation in hyaluronic acid/collagen hydrogel. Process Biochem. 50:2242–2250. 2015. View Article : Google Scholar
33	Blanco FJ, Geng Y and Lotz M: Differentiation-dependent effects of IL-1 and TGF-beta on human articular chondrocyte proliferation are related to inducible nitric oxide synthase expression. J Immunol. 154:4018–4026. 1995.PubMed/NCBI
34	Roberts AB, Flanders KC, Heine UI, Jakowlew S, Kondaiah P, Kim SJ and Sporn MB: Transforming growth factor-beta: multifunctional regulator of differentiation and development. Philos Trans R Soc Lond B Biol Sci. 327:145–154. 1990. View Article : Google Scholar : PubMed/NCBI
35	Wang P, Zhang F, He Q, Wang J, Shiu HT, Shu Y, Tsang WP, Liang S, Zhao K and Wan C: Flavonoid compound icariin activates hypoxia inducible factor-1α in chondrocytes and promotes articular cartilage repair. PLoS One. 11:e01483722016. View Article : Google Scholar
36	Shantikumar S, Caporali A and Emanueli C: Role of microRNAs in diabetes and its cardiovascular complications. Cardiovasc Res. 93:583–593. 2012. View Article : Google Scholar :
37	Schoolmeesters A, Eklund T, Leake D, Vermeulen A, Smith Q, Force Aldred S and Fedorov Y: Functional profiling reveals critical role for miRNA in differentiation of human mesenchymal stem cells. PLoS One. 4:e56052009. View Article : Google Scholar : PubMed/NCBI
38	Bueno MJ, Pérez de Castro I and Malumbres M: Control of cell proliferation pathways by microRNAs. Cell Cycle. 7:3143–3148. 2008. View Article : Google Scholar : PubMed/NCBI
39	Sumiyoshi K, Kubota S, Ohgawara T, Kawata K, Abd El Kader T, Nishida T, Ikeda N, Shimo T, Yamashiro T and Takigawa M: Novel role of miR-181a in cartilage metabolism. J Cell Biochem. 114:2094–2100. 2013. View Article : Google Scholar : PubMed/NCBI
40	Mirzamohammadi F, Papaioannou G and Kobayashi T: MicroRNAs in cartilage development, homeostasis, and disease. Curr Osteoporos Rep. 12:410–419. 2014. View Article : Google Scholar : PubMed/NCBI
41	Guérit D, Brondello JM, Chuchana P, Philipot D, Toupet K, Bony C, Jorgensen C and Noël D: FOXO3A regulation by miRNA-29a controls chondrogenic differentiation of mesenchymal stem cells and cartilage formation. Stem Cells Dev. 23:1195–1205. 2014. View Article : Google Scholar : PubMed/NCBI
42	Zhang L, Zhang X, Li KF, Li DX, Xiao YM, Fan YJ and Zhang XD: Icariin promotes extracellular matrix synthesis and gene expression of chondrocytes in vitro. Phytother Res. 26:1385–1392. 2012. View Article : Google Scholar : PubMed/NCBI