Curcumin inhibits nuclear translocation of nuclear factor-kappa B P65 in a rat model of traumatic osteoarthritis

doi:10.3969/j.issn.2095-4344.2016.15.005

Abstract

Abstract:

BACKGROUND: Mechanical, inflammatory, and biochemical factors, particularly matrix metalloproteinases and reactive oxygen lead to chondrocyte degeneration in osteoarthritis. Curcumin has been shown to be a potent antioxidant; however, its protective effects against chondrocyte degeneration in osteoarthritis remain unclear.
OBJECTIVE: To investigate the potential molecular mechanisms underlying the protective effects of curcumin on articular cartilage of osteoarthritis in rats.
METHODS: A total of 30 Sprague-Dawley rats were used and randomly divided into model group (positive control, n=15) and normal group (negative control, n=15). Rat models of traumatic osteoarthritis were established, and then cartilage cells were isolated from articular cartilage and cultured in vitro. Chondrocytes were treated with curcumin (curcumin group) or PDTC (an inhibitor of nuclear factor-kappa B) for 24 hours. The expression level of nuclear factor-kappa B P65 in nucleus and cytoplasm in chondrocytes were determined by western blot assay and immunofluorescence. Moreover, mRNA expressions of type II collagen, matrix metalloproteinase-1 and -13 were analyzed using RT-qPCR.
RESULTS AND CONCLUSION: Nuclear factor-kappa B P65 protein was mainly expressed in nucleus, but few in cytoplasm in positive control group; the reversed results were found in the curcumin group. Nuclear translocation of nuclear factor-kappa B P65 was observed mainly in nucleus in the positive control group; however, that was observed mainly in cytoplasm in the negative control, curcumin, and PDTC groups. Matrix metalloproteinase-1 and -13 mRNA expressions were significantly decreased, while type II collagen mRNA expression was significantly increased in the curcumin group compared with the positive control group. These findings indicated that curcumin protect chondrocytes against degeneration through inhibiting the activation of nuclear factor-kappa B signaling pathway, suppressing nuclear translocation of nuclear factor-kappa B P65 and inhibiting the expressions of matrix metalloproteinase-1 and -13, which are responsible for upregulation of type II collagen expression.
中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Curcumin, Osteoarthritis, NF-kappa B, Matrix Metalloproteinases, Collagen Type II, Tissue Engineering

Wang Jian, Ma Jie, Gu Jian-hua, Wang Fu-yong, Shang Xiu-shuai, Wang Zhao-fei, Wang Xiang, Tao Hai-rong. Curcumin inhibits nuclear translocation of nuclear factor-kappa B P65 in a rat model of traumatic osteoarthritis[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(15): 2163-2170.

Figures/Tables 5

References

[1] Shi D,Dai J,Xu Z,et al.Update on basic and clinical aspects of osteoarthritis. Ann Transl Med. 2015;3(10):142.

[2] Abeles AM, Pillinger MH.The role of the synovial fibroblast in rheumatoid arthritis: cartilage destruction and the regulation of matrix metalloproteinases. Bull NYU Hosp Jt Dis. 2006;64(1-2):20-24.

[3] Tchetverikov I,Lohmander LS,Verzijl N,et al. MMP protein and activity levels in synovial fluid from patients with joint injury, inflammatory arthritis, and osteoarthritis. Ann Rheum Dis. 2005;64(5):694-698.

[4] Aigner T,Soder S,Gebhard PM,et al.Mechanisms of disease: role of chondrocytes in the pathogenesis of osteoarthritis--structure, chaos and senescence. Nat Clin Pract Rheumatol. 2007;3(7):391-399.

[5] Pardo A,Selman M.MMP-1: the elder of the family. Int J Biochem Cell Biol.2005; 37(2):283-288.

[6] Tardif G,Reboul P, Pelletier JP, et al.Ten years in the life of an enzyme: the story of the human MMP-13 (collagenase-3).Mod Rheumatol. 2004;14(3):197-204.

[7] Bharti AC, Aggarwal BB.Nuclear factor-kappa B and cancer: its role in prevention and therapy. Biochemical pharmacology.2002;64(5-6):883-888.

[8] Mukhopadhyay A, Bueso-Ramos C, Chatterjee D,et al. Curcumin downregulates cell survival mechanisms in human prostate cancer cell lines.Oncogene.2001; 20(52):7597-7609.

[9] Bharti AC,Donato N,Singh S,et al.Curcumin (diferuloylmethane) down-regulates the constitutive activation of nuclear factor-kappa B and IkappaBalpha kinase in human multiple myeloma cells, leading to suppression of proliferation and induction of apoptosis. Blood.2003;101(3):1053-1062.

[10] Wan XH,Li YW,Luo XP. Curcumin attenuated the lipid peroxidation and apoptotic liver injury in copper-overloaded rats. Zhonghua Er Ke Za Zhi. 2007;45(8):604-608.

[11] Shen SQ,Zhang Y,Xiang JJ,et al.Protective effect of curcumin against liver warm ischemia/reperfusion injury in rat model is associated with regulation of heat shock protein and antioxidant enzymes. World J Gastroenterol. 2007;13(13):1953-1961.

[12] Chen Q,Wang Y,Xu K,et al.Curcumin induces apoptosis in human lung adenocarcinoma A549 cells through a reactive oxygen species-dependent mitochondrial signaling pathway. Oncology reports.2010;23(2):397-403.

[13] Zhang XH, Xu XX, Xu T.Ginsenoside Ro suppresses interleukin-1beta-induced apoptosis and inflammation in rat chondrocytes by inhibiting NF-kappaB. Chin J Nat Med.2015;13(4):283-289.

[14] Lee WT,Lee TH,Cheng CH,et al.Antroquinonol from Antrodia Camphorata suppresses breast tumor migration/invasion through inhibiting ERK-AP-1- and AKT-NF-kappaB-dependent MMP-9 and epithelial-mesenchymal transition expressions. Food Chem Toxicol. 2015;78:33-7841.

[15] Wang JJ, Huan SK, Hsieh KH, et al. Inhibitory effect of midazolam on MMP-9, MMP-1 and MMP-13 expression in PMA-stimulated human chondrocytes via recovery of NF-kappaB signaling. Arch Med Sci. 2013;9(2):332-339.

[16] Liacini A,Sylvester J,Li WQ,et al.Induction of matrix metalloproteinase-13 gene expression by TNF-alpha is mediated by MAP kinases, AP-1, and NF-kappaB transcription factors in articular chondrocytes.Exp Cell Res. 2003;288(1):208-217.

[17] Chen C,Ma C,Zhang Y,et al.Pioglitazone inhibits advanced glycation end product-induced TNF-alpha and MMP-13 expression via the antagonism of NF-kappaB activation in chondrocytes. Pharmacology. 2014;94(5-6):265-272.

[18] Cheng CC, Chen YH, Chang WL,et al.Phytoestrogen bavachin mediates anti-inflammation targeting Ikappa B kinase-I kappaB alpha-NF-kappaB signaling pathway in chondrocytes in vitro. Eur J Pharmacol. 2010;636(1-3):181-188.

[19] Caterson B,Flannery CR,Hughes CE,et al. Mechanisms involved in cartilage proteoglycan catabolism. Matrix biology: Matrix Biol. 2000;19(4):333-344.

[20] Yoshihara Y,Nakamura H,Obata K,et al.Matrix metalloproteinases and tissue inhibitors of metalloproteinases in synovial fluids from patients with rheumatoid arthritis or osteoarthritis. Ann Rheum Dis. 2000;59(6):455-461.

[21] Westermarck J,Kahari VM.Regulation of matrix metalloproteinase expression in tumor invasion. FASEB J.1999;13(8):781-792.

[22] Mort JS, Billington CJ.Articular cartilage and changes in arthritis: matrix degradation. Arthritis Res.2001;3(6): 337-341.

[23] Goldring SR,Goldring MB.The role of cytokines in cartilage matrix degeneration in osteoarthritis. Clin Orthop Relat Res. 2004;(427 Suppl):S27-36.

[24] Shakibaei M,John T,Schulze-Tanzil G,et al.Suppression of NF-kappaB activation by curcumin leads to inhibition of expression of cyclo-oxygenase-2 and matrix metalloproteinase-9 in human articular chondrocytes: Implications for the treatment of osteoarthritis.Biochem Pharmacol. 2007;73(9):1434-1445.