UROTENSİN II’NİN İNSAN OSTEOBLAST HÜCRELERİNDE ENFLAMASYONLA İLİŞKİLİ ROLÜ

Yıl 2021, , 353 - 361, 25.03.2021

Hamza Malik Okuyan , Menderes Yusuf Terzi , Gülay Duran , Aydıner Kalacı

https://doi.org/10.33715/inonusaglik.834716

Cited By: 1

Öz

Osteoartrit (OA), kemik, kıkırdak ve sinoviyal dokuda katabolik değişikliklere neden olan yaygın bir eklem hastalığıdır. OA’nın güncel tedavisi, hastalığı tamamen iyileştirmek için yeterli değildir. Eklem kondrositlerinin yanı sıra, kemik hücrelerindeki OA ile ilişkili mekanizmaların anlaşılması bu hastalığın önlenmesi ve tedavisine katkı sağlayacaktır. Urotensin II (UTS2), nörohormon benzeri aktiviteye sahip polipeptid bir moleküldür ve kardiyovasküler, pulmoner, renal ve merkezi sinir sisteminde eksprese edildiği bilinmektedir. Pek çok sistem üzerinde patofizyolojik etkileri olan UTS2’nin, OA’daki moleküler mekanizması tam olarak bilinmemektedir. Bu çalışmada, insan osteoblast hücreleri kullanılarak UTS2’nin OA patogenezindeki enflamasyonla ve apoptozis ile ilişkili rolünün incelenmesi amaçlanmıştır. Bu çalışmada, insan osteoblast hücreleri, tümör nekroz faktörü alfa (TNF-α) ve UTS2’ye maruz bırakıldı. İnkübasyon süresinin sonunda hücrelerden RNA izole edildi ve kantitatif ters transkripsiyon-polimeraz zincir reaksiyonu analizleri yapıldı. TNF-α maruziyeti, interlökin 6 ve nükleer faktör kappa B gen ekspresyon seviyelerinde anlamlı düzeyde bir artışa neden olmuştur. UTS2 maruziyeti, kendi gen ekspresyon seviyesini azaltmıştır. TNF-a ve UTS2 maruziyetleri, Kaspaz-3 gen ekspresyonu üzerinde istatistiksel olarak anlamlı bir farklılığa neden olmamıştır. Çalışmamızda, bulgularımız; insan osteoblast hücrelerinde UTS2’nin eksprese edildiğini ve UTS2’nin hücre kültür ortamına eklenmesi bu genin kendi ekspresyon seviyeleri üzerinde etkili olabileceğini göstermektedir.

Anahtar Kelimeler

Osteoartrit, Urotensin II, TNF-α, IL-6, Osteoblast

Destekleyen Kurum

Bu çalışma Hatay Mustafa Kemal Üniversitesi Bilimsel Araştırmalar Projeler Birimi tarafından desteklenmiştir.

Proje Numarası

18.M.089

The Role of Urotensin II Associated with Inflammation in Human Osteoblast Cells

Öz

Osteoarthritis (OA) is a common joint disease that causes catabolic changes in bone, cartilage and synovial tissue. The current treatment of OA is not sufficient to completely cure the disease. Understanding of OA-related mechanisms in bone cells as well as in joint chondrocytes will contribute to the prevention and treatment of this disease. Urotensin II (UTS2) is a polypeptide molecule with neurohormone-like activity and is known to be expressed in the cardiovascular, pulmonary, renal, and central nervous system. The molecular mechanism of UTS2, which has pathophysiological effects on many systems, in OA is not fully known. In this study, it has been aimed to investigate the role of UTS2 associated with inflammation and apoptosis in OA pathogenesis by using human osteoblast cells. In this study, human osteoblast cells were exposed to tumor necrosis factor alpha (TNF-α) and UTS2. At the end of the incubation period, RNA was isolated from the cells and quantitative real-time polymerase chain reaction analyses were performed. TNF-α treatment has caused a significant increase in interleukin 6 and nuclear factor kappa B gene expression levels. UTS2 treatment has reduced its own gene expression level. TNF-α and UTS2 treatments have not caused a statistically significant difference in Caspase-3 gene expression. In our study, our findings show that UTS2 is expressed in human osteoblast cells and the addition of UTS2 into the cell culture medium can affect its own expression levels.

Anahtar Kelimeler

Osteoarthritis, Urotensin II, TNF-α, IL-6, Osteoblast

Proje Numarası

18.M.089

Kaynakça

• Ames, R. S., Sarau, H. M., Chambers, J. K., Willette, R. N., Aiyar, N. V., Romanic, A. M., … Douglas, S.A. (1999). Human urotensin-II is a potent vasoconstrictor and agonist for the orphan receptor GPR14. Nature, 401(6750), 282-286. doi: 10.1038/45809.
• Berlind, A. (1972). Teleost caudal neurosecretory system: release of urotensin II from isolated urophyses. Gen Comp Endocrinol, 18(3), 557-560. doi: 10.1016/0016-6480(72)90036-6.
• Bin, G., Cuifang, W., Bo, Z., Jing, W., Jin, J., Xiaoyi, T., …Yayi, X. (2015). Fluid shear stress inhibits TNF-alpha-induced osteoblast apoptosis via ERK5 signaling pathway. Biochem Biophys Res Commun, 466(1), 117-123. doi: 10.1016/j.bbrc.2015.08.117
• Davenport, A. P., Maguire, J. J. (2000). Urotensin II: fish neuropeptide catches orphan receptor. Trends Pharmacol Sci, 21(3), 80-82.
• Denizot, F., Lang, R. (1986). Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J Immunol Methods, 89(2), 271-277. doi: 10.1016/0022-1759(86)90368-6
• Gogebakan, B., Uruc, V., Ozden, R., Duman, I. G., Yagiz, A. E., Okuyan, H. M., ...Kalacı, A. (2014). Urotensin II (U-II), a novel cyclic peptide, possibly associated with the pathophysiology of osteoarthritis. Peptides, 54, 159-161. doi: 10.1016/j.peptides.2014.01.010
• Goldring, M. B., Goldring, S. R. (2010). Articular cartilage and subchondral bone in the pathogenesis of osteoarthritis. Ann N Y Acad Sci, 1192, 230-237. doi: 10.1111/j.1749-6632.2009.05240.x
• Hassan, G. S., Douglas, S. A., Ohlstein, E. H., Giaid, A. (2005). Expression of urotensin-II in human coronary atherosclerosis. Peptides, 26(12), 2464-2472. doi: 10.1016/j.peptides.2005.05.028
• Holt, I., Davie, M. W., Marshall, M. J. (1996). Osteoclasts are not the major source of interleukin-6 in mouse parietal bones. Bone, 18(3), 221-226. doi: 10.1016/8756-3282(95)00482-3
• Ishimi, Y., Miyaura, C., Jin, C. H., Akatsu, T., Abe, E., Nakamura, Y., ... Hirano, T. (1990). IL-6 is produced by osteoblasts and induces bone resorption. J Immunol, 145(10), 3297-3303.
• Kotake, S., Sato, K., Kim, K. J., Takahashi, N., Udagawa, N., Nakamura, I., …Kashiwazaki, S. (1996). Interleukin-6 and soluble interleukin-6 receptors in the synovial fluids from rheumatoid arthritis patients are responsible for osteoclast-like cell formation. J Bone Miner Res, 11(1), 88-95. doi: 10.1002/jbmr.5650110113
• Liu, L. M., Liang, D. Y., Ye, C. G., Tu, W. J.,Zhu, T. (2015). The UII/UT system mediates upregulation of proinflammatory cytokines through p38 MAPK and NF-kappaB pathways in LPS-stimulated Kupffer cells. PLoS One, 10(3), e0121383. doi: 10.1371/journal.pone.0121383
• Livak, K. J., Schmittgen, T. D. (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods, 25(4), 402-408. doi: 10.1006/meth.2001.1262
• Menetrey, J., Unno-Veith, F., Madry, H., Van Breuseghem, I. (2010). Epidemiology and imaging of the subchondral bone in articular cartilage repair. Knee Surg Sports Traumatol Arthrosc, 18(4), 463-471. doi: 10.1007/s00167-010-1053-0
• Mohan, G., Perilli, E., Kuliwaba, J. S., Humphries, J. M., Parkinson, I. H., Fazzalari, N. L. (2011). Application of in vivo micro-computed tomography in the temporal characterisation of subchondral bone architecture in a rat model of low-dose monosodium iodoacetate-induced osteoarthritis. Arthritis Res Ther, 13(6), R210. doi: 10.1186/ar3543
• Okumus, S., Igci, Y. Z., Taskin, T., Oztuzcu, S., Gurler, B., Eslik, Z., …Demiryurek, A.T. (2012). Association between Thr21Met and Ser89Asn polymorphisms of the urotensin-II (UTS2) gene, diabetes mellitus, and diabetic retinopathy. Curr Eye Res, 37(10), 921-929. doi: 10.3109/02713683.2012.688181
• Pearson, D., Shively, J. E., Clark, B. R., Geschwind, II, Barkley, M., Nishioka, R. S., Bern, H.A. (1980). Urotensin II: a somatostatin-like peptide in the caudal neurosecretory system of fishes. Proc Natl Acad Sci U S A, 77(8), 5021-5024.
• Pehlivan, Y., Gogebakan, B., Oztuzcu, S., Ozgen, M., Cetin, G. Y., Bayraktar, R., …Onat, A.M.(2012). Association between Thr21Met and Ser89Asn polymorphisms of the urotensin II gene and systemic sclerosis. J Rheumatol, 39(1), 106-111. doi: 10.3899/jrheum.110509
• Segain, J. P., Rolli-Derkinderen, M., Gervois, N., Raingeard de la Bletiere, D., Loirand, G., Pacaud, P. (2007). Urotensin II is a new chemotactic factor for UT receptor-expressing monocytes. J Immunol, 179(2), 901-909. doi: 10.4049/jimmunol.179.2.901
• Sun, S. L., Liu, L. M. (2019). Urotensin II: an inflammatory cytokine. J Endocrinol. doi: 10.1530/JOE-18-0505
• Tomiyama, S., Nakamachi, T., Uchiyama, M., Matsuda, K.,Konno, N. (2015). Urotensin II upregulates migration and cytokine gene expression in leukocytes of the African clawed frog, Xenopus laevis. Gen Comp Endocrinol, 216, 54-63. doi: 10.1016/j.ygcen.2015.04.009
• Ugan, R. A., Cadirci, E., Halici, Z., Toktay, E., Cinar, I. (2018). The role of urotensin-II and its receptors in sepsis-induced lung injury under diabetic conditions. Eur J Pharmacol, 818, 457-469. doi: 10.1016/j.ejphar.2017.11.011
• Valverde-Franco, G., Pelletier, J. P., Fahmi, H., Hum, D., Matsuo, K., Lussier, B., ... Martel-Pelletier, J. (2012). In vivo bone-specific EphB4 overexpression in mice protects both subchondral bone and cartilage during osteoarthritis. Arthritis Rheum, 64(11), 3614-3625. doi: 10.1002/art.34638
• Yang, Y., Zhang, J., Chen, X., Wu, T., Xu, X., Cao, G., ...Li, Y. (2016). UII/GPR14 is involved in NF-kappaB-mediated colonic inflammation in vivo and in vitro. Oncol Rep, 36(5), 2800-2806. doi: 10.3892/or.2016.5069
• Yu, D., Xu, J., Liu, F., Wang, X., Mao, Y., Zhu, Z. (2016). Subchondral bone changes and the impacts on joint pain and articular cartilage degeneration in osteoarthritis. Clin Exp Rheumatol, 34(5), 929-934.