MicroRNA-34a and MicroRNA-181a Mediate Visfatin-Induced Apoptosis and Oxidative Stress via NF-κB Pathway in Human Osteoarthritic Chondrocytes
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chondrocyte Cultures
2.2. Treatment of Chondrocyte Cultures
2.3. Transfection of Chondrocytes
2.4. Detection of Apoptosis
2.5. RNA Isolation and Quantitative Real-Time PCR
2.6. Mitochondrial Superoxide Anion (•O2−) Production
2.7. Western Blot
2.8. Immunofluorescence Analysis
2.9. Statistical Analysis
3. Results
3.1. Visfatin Regulates Apoptosis and Oxidative Stress Processes
3.2. MiRNA Specific Inhibitors Block Visfatin Effect on Mir-34a and Mir-181a Gene Expression
3.3. MiR-34a and Mir-181a Specific Inhibitors Prevent Chondrocyte Apoptosis Induced by Visfatin by Regulating BCL2
3.4. MiR-34a and MiR-181a Specific Inhibitors Regulate Oxidative Stress Induced by Visfatin
3.5. NF-κB Pathway Mediates Visfatin Effects
3.6. MiR-34a and MiR-181a Regulate NF-κB Pathway
4. Discussion
5. Conclusions
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- Malemud, C.J. MicroRNAs and Osteoarthritis. Cells 2018, 7, 92. [Google Scholar] [CrossRef] [PubMed]
- Díaz-Prado, S.; Cicione, C.; Muiños-López, E.; Hermida-Gómez, T.; Oreiro, N.; Fernández-López, C.; Blanco, F.J. Characterization of microRNA expression profiles in normal and osteoarthritic human chondrocytes. BMC Musculoskelet. Disord. 2012, 13, 144. [Google Scholar] [CrossRef] [PubMed]
- De Palma, A.; Cheleschi, S.; Pascarelli, N.A.; Tenti, S.; Galeazzi, M.; Fioravanti, A. Do MicroRNAs have a key epigenetic role in osteoarthritis and in mechanotransduction? Clin. Exp. Rheumatol. 2017, 35, 518–526. [Google Scholar] [PubMed]
- Cheleschi, S.; De Palma, A.; Pascarelli, N.A.; Giordano, N.; Galeazzi, M.; Tenti, S.; Fioravanti, A. Could Oxidative Stress Regulate the Expression of MicroRNA-146a and MicroRNA-34a in Human Osteoarthritic Chondrocyte Cultures? Int. J. Mol. Sci. 2017, 18, 2660. [Google Scholar] [CrossRef] [PubMed]
- Bu, H.; Wedel, S.; Cavinato, M.; Jansen-Dürr, P. MicroRNA Regulation of Oxidative Stress-Induced Cellular Senescence. Oxidative Med. Cell. Longev. 2017, 2017, 1–12. [Google Scholar] [CrossRef]
- Neumann, E.; Junker, S.; Schett, G.; Frommer, K.; Müller-Ladner, U. Adipokines in bone disease. Nat. Rev. Rheumatol. 2016, 12, 296–302. [Google Scholar] [CrossRef] [PubMed]
- Francisco, V.; Pérez, T.; Pino, J.; López, V.; Franco, E.; Alonso, A.; Gonzalez-Gay, M.A.; Mera, A.; Lago, F.; Gómez, R.; et al. Biomechanics, obesity, and osteoarthritis. The role of adipokines: When the levee breaks. J. Orthop. Res. 2017, 36, 594–604. [Google Scholar] [CrossRef] [Green Version]
- Tenti, S.; Palmitesta, P.; Giordano, N.; Galeazzi, M.; Fioravanti, A. Increased serum leptin and visfatin levels in patients with diffuse idiopathic skeletal hyperostosis: A comparative study. Scand. J. Rheumatol. 2017, 46, 156–158. [Google Scholar] [CrossRef]
- Fioravanti, A.; Cheleschi, S.; De Palma, A.; Addimanda, O.; Mancarella, L.; Pignotti, E.; Pulsatelli, L.; Galeazzi, M.; Meliconi, R. Can adipokines serum levels be used as biomarkers of hand osteoarthritis? Biomarkers 2018, 23, 265–270. [Google Scholar] [CrossRef]
- Francisco, V.; Ruiz-Fernández, C.; Pino, J.; Mera, A.; Gonzalez-Gay, M.A.; Gómez, R.; Lago, F.; Mobasheri, A.; Gualillo, O. Adipokines: Linking metabolic syndrome, the immune system, and arthritic diseases. Biochem. Pharmacol. 2019, 165, 196–206. [Google Scholar] [CrossRef]
- Fioravanti, A.; Giannitti, C.; Cheleschi, S.; Simpatico, A.; Pascarelli, N.A.; Galeazzi, M. Circulating levels of adiponectin, resistin, and visfatin after mud-bath therapy in patients with bilateral knee osteoarthritis. Int. J. Biometeorol. 2015, 59, 1691–1700. [Google Scholar] [CrossRef] [PubMed]
- Wu, M.-H.; Tsai, C.-H.; Huang, Y.-L.; Fong, Y.-C.; Tang, C.-H. Visfatin Promotes IL-6 and TNF-α Production in Human Synovial Fibroblasts by Repressing miR-199a-5p through ERK, p38 and JNK Signaling Pathways. Int. J. Mol. Sci. 2018, 19, 190. [Google Scholar] [CrossRef] [PubMed]
- Cheleschi, S.; Giordano, N.; Volpi, N.; Tenti, S.; Gallo, I.; Di Meglio, M.; Giannotti, S.; Fioravanti, A. A Complex Relationship between Visfatin and Resistin and microRNA: An In Vitro Study on Human Chondrocyte Cultures. Int. J. Mol. Sci. 2018, 19, 3909. [Google Scholar] [CrossRef] [PubMed]
- Maurizi, G.; Babini, L.; Della Guardia, L. Potential role of microRNAs in the regulation of adipocytes liposecretion and adipose tissue physiology. J. Cell. Physiol. 2018, 233, 9077–9086. [Google Scholar] [CrossRef] [PubMed]
- Altman, R.; Alarcon, G.; Appelrouth, D.; Bloch, D.; Borenstein, D.; Brandt, K.; Brown, C.; Cooke, T.D.; Daniel, W.; Feldman, D.; et al. The American College of Rheumatology criteria for the classification and reporting of osteoarthritis of the hip. Arthritis Rheum. 1991, 34, 505–514. [Google Scholar] [CrossRef] [PubMed]
- Mankin, H.J.; Dorfman, H.; Lippiello, L.; Zarins, A. Biochemical and metabolic abnormalities in articular cartilage from osteo-arthritic human hips. II. Correlation of morphology with biochemical and metabolic data. J. Bone Jt. Surgery-American Vol. 1971, 53, 523–537. [Google Scholar] [CrossRef]
- Francin, P.-J.; Guillaume, C.; Humbert, A.-C.; Pottie, P.; Netter, P.; Mainard, D.; Presle, N. Association between the chondrocyte phenotype and the expression of adipokines and their receptors: Evidence for a role of leptin but not adiponectin in the expression of cartilage-specific markers. J. Cell. Physiol. 2011, 226, 2790–2797. [Google Scholar] [CrossRef]
- Gosset, M.; Berenbaum, F.; Salvat, C.; Sautet, A.; Pigenet, A.; Tahiri, K.; Jacques, C. Crucial role of visfatin/pre–B cell colony-enhancing factor in matrix degradation and prostaglandin E2 synthesis in chondrocytes: Possible influence on osteoarthritis. Arthritis Rheum. 2008, 58, 1399–1409. [Google Scholar] [CrossRef]
- Cheleschi, S.; De Palma, A.; Pecorelli, A.; Pascarelli, N.A.; Valacchi, G.; Belmonte, G.; Carta, S.; Galeazzi, M.; Fioravanti, A. Hydrostatic Pressure Regulates MicroRNA Expression Levels in Osteoarthritic Chondrocyte Cultures via the Wnt/β-Catenin Pathway. Int. J. Mol. Sci. 2017, 18, 133. [Google Scholar] [CrossRef]
- Mobasheri, A.; Rayman, M.P.; Gualillo, O.; Sellam, J.; Van Der Kraan, P.; Fearon, U. The role of metabolism in the pathogenesis of osteoarthritis. Nat. Rev. Rheumatol. 2017, 13, 302–311. [Google Scholar] [CrossRef]
- Deiuliis, J.A. MicroRNAs as regulators of metabolic disease: Pathophysiologic significance and emerging role as biomarkers and therapeutics. Int. J. Obes. 2016, 40, 88–101. [Google Scholar] [CrossRef] [PubMed]
- Lei, L.; Zhou, C.; Yang, X.; Li, L. Down-regulation of microRNA-375 regulates adipokines and inhibits inflammatory cytokines by targeting AdipoR2 in non-alcoholic fatty liver disease. Clin. Exp. Pharmacol. Physiol. 2018, 45, 819–831. [Google Scholar] [CrossRef] [PubMed]
- Jasinski-Bergner, S.; Kielstein, H. Adipokines Regulate the Expression of Tumor-Relevant MicroRNAs. Obes. Facts 2019, 12, 211–225. [Google Scholar] [CrossRef] [PubMed]
- Musumeci, G.; Castrogiovanni, P.; Trovato, F.M.; Weinberg, A.M.; Al-Wasiyah, M.K.; Alqahtani, M.H.; Mobasheri, A. Biomarkers of Chondrocyte Apoptosis and Autophagy in Osteoarthritis. Int. J. Mol. Sci. 2015, 16, 20560–20575. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sun, L.; Chen, S.; Gao, H.; Ren, L.; Song, G. Visfatin induces the apoptosis of endothelial progenitor cells via the induction of pro-inflammatory mediators through the NF-κB pathway. Int. J. Mol. Med. 2017, 40, 637–646. [Google Scholar] [CrossRef]
- Yan, S.; Wang, M.; Zhao, J.; Zhang, H.; Zhou, C.; Jin, L.; Zhang, Y.; Qiu, X.; Ma, B.; Fan, Q. MicroRNA-34a affects chondrocyte apoptosis and proliferation by targeting the SIRT1/p53 signaling pathway during the pathogenesis of osteoarthritis. Int. J. Mol. Med. 2016, 38, 201–209. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Abouheif, M.M.; Nakasa, T.; Shibuya, H.; Niimoto, T.; Kongcharoensombat, W.; Ochi, M. Silencing microRNA-34a inhibits chondrocyte apoptosis in a rat osteoarthritis model in vitro. Rheumatology 2010, 49, 2054–2060. [Google Scholar] [CrossRef] [Green Version]
- Bartel, D.P.; Chen, C.-Z.; Li, L.; Lodish, H.F. MicroRNAs Modulate Hematopoietic Lineage Differentiation. Science 2004, 303, 83–86. [Google Scholar] [Green Version]
- Okuhara, A.; Nakasa, T.; Shibuya, H.; Niimoto, T.; Adachi, N.; Deie, M.; Ochi, M. Changes in microRNA expression in peripheral mononuclear cells according to the progression of osteoarthritis. Mod. Rheumatol. 2012, 22, 446–457. [Google Scholar] [CrossRef]
- Nakamura, A.; Rampersaud, Y.R.; Sharma, A.; Lewis, S.J.; Wu, B.; Datta, P.; Sundararajan, K.; Endisha, H.; Rossomacha, E.; Rockel, J.S.; et al. Identification of microRNA-181a-5p and microRNA-4454 as mediators of facet cartilage degeneration. JCI Insight 2016, 1, 86820. [Google Scholar] [CrossRef]
- De Palma, A.; Cheleschi, S.; Pascarelli, N.A.; Giannotti, S.; Galeazzi, M.; Fioravanti, A. Hydrostatic pressure as epigenetic modulator in chondrocyte cultures: A study on miRNA-155, miRNA-181a and miRNA-223 expression levels. J. Biomech. 2018, 66, 165–169. [Google Scholar] [CrossRef] [PubMed]
- Zheng, H.; Liu, J.; Tycksen, E.; Nunley, R.; McAlinden, A. MicroRNA-181a/b-1 over-expression enhances osteogenesis by modulating PTEN/PI3K/AKT signaling and mitochondrial metabolism. Bone 2019, 123, 92–102. [Google Scholar] [CrossRef] [PubMed]
- Feng, X.; Zhang, C.; Yang, Y.; Hou, D.; Zhu, A. Role of miR-181a in the process of apoptosis of multiple malignant tumors: A literature review. Adv. Clin. Exp. Med. 2018, 27, 263–270. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Nakamura, A.; Rampersaud, Y.R.; Nakamura, S.; Sharma, A.M.; Zeng, F.; Rossomacha, E.; Ali, S.A.; Krawetz, R.; Haroon, N.; Perruccio, A.V.; et al. microRNA-181a-5p antisense oligonucleotides attenuate osteoarthritis in facet and knee joints. Ann. Rheum. Dis. 2019, 78, 111–121. [Google Scholar] [CrossRef] [PubMed]
- Kim, D.; Song, J.; Kim, S.; Chun, C.-H.; Jin, E.-J. MicroRNA-34a regulates migration of chondroblast and IL-1β-induced degeneration of chondrocytes by targeting EphA5. Biochem. Biophys. Res. Commun. 2011, 415, 551–557. [Google Scholar] [CrossRef] [PubMed]
- Fisch, K.M.; Akagi, R.; Alvarez-Garcia, O.; Teramura, T.; Muramatsu, Y.; Saito, M.; Duffy, S.; Grogan, S.; Sasho, T.; D’Lima, D.; et al. Integrative omics profiling reveals dysregulated novel pathways mediated by microRNAs and DNA methylation in osteoarthritis. Arthritis Rheumatol. 2014, 66, S829–S830. [Google Scholar]
- Ouyang, Y.B.; Lu, Y.; Yue, S.; Giffard, R.G. miR-181 targets multiple Bcl-2 family members and influences apoptosis and mitochondrial function in astrocytes. Mitochondrion 2012, 12, 213–219. [Google Scholar] [CrossRef] [Green Version]
- D’Adamo, S.; Cetrullo, S.; Guidotti, S.; Borzì, R.M.; Flamigni, F.; D’Adamo, S. Hydroxytyrosol modulates the levels of microRNA-9 and its target sirtuin-1 thereby counteracting oxidative stress-induced chondrocyte death. Osteoarthr. Cartil. 2017, 25, 600–610. [Google Scholar] [CrossRef] [Green Version]
- Davies, C.M.; Guilak, F.; Weinberg, J.B.; Fermor, B. Reactive nitrogen and oxygen species in interleukin-1-mediated DNA damage associated with osteoarthritis. Osteoarthr. Cartil. 2008, 16, 624–630. [Google Scholar] [CrossRef] [Green Version]
- Teixeira, T.M.; Resende, A.C.; Bezerra, F.F.; Da Costa, D.C.; O Soulage, C.; Daleprane, J.B. Activation of Nrf2-Antioxidant Signaling by 1,25-Dihydroxycholecalciferol Prevents Leptin-Induced Oxidative Stress and Inflammation in Human Endothelial Cells. J. Nutr. 2017, 147, 506–513. [Google Scholar] [CrossRef]
- Khan, N.M.; Ahmad, I.; Haqqi, T.M. Nrf2/ARE pathway attenuates oxidative and apoptotic response in human osteoarthritis chondrocytes by activating ERK1/2/ELK1-P70S6K-P90RSK signaling axis. Free. Radic. Boil. Med. 2018, 116, 159–171. [Google Scholar] [CrossRef]
- Mathy-Hartert, M.; Hogge, L.; Sanchez, C.; Deby-Dupont, G.; Crielaard, J.; Henrotin, Y. Interleukin-1β and interleukin-6 disturb the antioxidant enzyme system in bovine chondrocytes: a possible explanation for oxidative stress generation. Osteoarthr. Cartil. 2008, 16, 756–763. [Google Scholar] [CrossRef]
- Huang, M.L.-H.; Chiang, S.; Kalinowski, D.S.; Bae, D.-H.; Sahni, S.; Richardson, D.R. The Role of the Antioxidant Response in Mitochondrial Dysfunction in Degenerative Diseases: Cross-Talk between Antioxidant Defense, Autophagy, and Apoptosis. Oxidative Med. Cell. Longev. 2019, 2019, 1–26. [Google Scholar] [CrossRef] [Green Version]
- McMahon, M.; Thomas, N.; Itoh, K.; Yamamoto, M.; Hayes, J.D. Redox-regulated Turnover of Nrf2 Is Determined by at Least Two Separate Protein Domains, the Redox-sensitive Neh2 Degron and the Redox-insensitive Neh6 Degron*. J. Boil. Chem. 2004, 279, 31556–31567. [Google Scholar] [CrossRef]
- Marchev, A.S.; Dimitrova, P.A.; Burns, A.J.; Kostov, R.V.; Dinkova-Kostova, A.T.; Georgiev, M.I. Oxidative stress and chronic inflammation in osteoarthritis: can NRF2 counteract these partners in crime? Ann. New York Acad. Sci. 2017, 1401, 114–135. [Google Scholar] [CrossRef]
- Wang, L.; Huang, H.; Fan, Y.; Kong, B.; Hu, H.; Hu, K.; Guo, J.; Mei, Y.; Liu, W.-L. Effects of Downregulation of MicroRNA-181a on H2O2-Induced H9c2 Cell Apoptosis via the Mitochondrial Apoptotic Pathway. Oxidative Med. Cell. Longev. 2014, 2014, 1–16. [Google Scholar] [CrossRef]
- Baker, J.R.; Vuppusetty, C.; Colley, T.; Papaioannou, A.I.; Fenwick, P.; Donnelly, L.; Ito, K.; Barnes, P.J. Oxidative stress dependent microRNA-34a activation via PI3Kα reduces the expression of sirtuin-1 and sirtuin-6 in epithelial cells. Sci. Rep. 2016, 6, 35871. [Google Scholar] [CrossRef]
- Chen, K.L.; Fu, Y.Y.; Shi, M.Y.; Li, H.X. Down-regulation of miR-181a can reduce heat stress damage in PBMCs of Holstein cows. In Vitro Cell Dev. Biol. Anim. 2016, 52, 864–871. [Google Scholar] [CrossRef]
- Zhong, X.; Li, P.; Li, J.; He, R.; Cheng, G.; Li, Y. Downregulation of microRNA-34a inhibits oxidized low-density lipoprotein-induced apoptosis and oxidative stress in human umbilical vein endothelial cells. Int. J. Mol. Med. 2018, 42, 1134–1144. [Google Scholar] [CrossRef]
- Rigoglou, S.; Papavassiliou, A.G. The NF-κB signalling pathway in osteoarthritis. Int. J. Biochem. Cell Boil. 2013, 45, 2580–2584. [Google Scholar] [CrossRef]
- Zhang, Q.; Lenardo, M.J.; Baltimore, D. 30 years of NF-κB: a blossoming of relevance to human pathobiology. Cell 2017, 168, 37–57. [Google Scholar] [CrossRef]
- Romacho, T.; Azcutia, V.; Vázquez-Bella, M.; Matesanz, N.; Cercas, E.; Nevado, J.; Carraro, R.; Rodriguez-Mañas, L.; Sánchez-Ferrer, C.F.; Peiro, C. Extracellular PBEF/NAMPT/visfatin activates pro-inflammatory signalling in human vascular smooth muscle cells through nicotinamide phosphoribosyltransferase activity. Diabetologia 2009, 52, 2455–2463. [Google Scholar] [CrossRef] [Green Version]
- Qi, J.; Qiao, Y.; Wang, P.; Li, S.; Zhao, W.; Gao, C. microRNA-210 negatively regulates LPS-induced production of proinflammatory cytokines by targeting NF-κB1 in murine macrophages. FEBS Lett. 2012, 586, 1201–1207. [Google Scholar] [CrossRef]
- Wen, F.; Yang, Y.; Jin, D.; Sun, J.; Yu, X.; Yang, Z. MiRNA-145 is involved in the development of resistin-induced insulin resistance in HepG2 cells. Biochem. Biophys. Res. Commun. 2014, 445, 517–523. [Google Scholar] [CrossRef]
- Shi, J.; Zhou, B.; Li, H. miR-27 inhibits the NF-κB signaling pathway by targeting leptin in osteoarthritic chondrocytes. Int. J. Mol. Med. 2017, 40, 523–530. [Google Scholar]
miRNA Genes | Cat. No. (Qiagen) |
miR-34a | MS00003318 |
miR-181a | MS00006692 |
SNORD-25 | MS00014007 |
Target Genes | Cat. No. (Qiagen) |
BCL2 | QT00000721 |
SOD-2 | QT01008693 |
CAT | QT00079674 |
NRF2 | QT00027384 |
ACTB | QT00095431 |
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Cheleschi, S.; Tenti, S.; Mondanelli, N.; Corallo, C.; Barbarino, M.; Giannotti, S.; Gallo, I.; Giordano, A.; Fioravanti, A. MicroRNA-34a and MicroRNA-181a Mediate Visfatin-Induced Apoptosis and Oxidative Stress via NF-κB Pathway in Human Osteoarthritic Chondrocytes. Cells 2019, 8, 874. https://doi.org/10.3390/cells8080874
Cheleschi S, Tenti S, Mondanelli N, Corallo C, Barbarino M, Giannotti S, Gallo I, Giordano A, Fioravanti A. MicroRNA-34a and MicroRNA-181a Mediate Visfatin-Induced Apoptosis and Oxidative Stress via NF-κB Pathway in Human Osteoarthritic Chondrocytes. Cells. 2019; 8(8):874. https://doi.org/10.3390/cells8080874
Chicago/Turabian StyleCheleschi, Sara, Sara Tenti, Nicola Mondanelli, Claudio Corallo, Marcella Barbarino, Stefano Giannotti, Ines Gallo, Antonio Giordano, and Antonella Fioravanti. 2019. "MicroRNA-34a and MicroRNA-181a Mediate Visfatin-Induced Apoptosis and Oxidative Stress via NF-κB Pathway in Human Osteoarthritic Chondrocytes" Cells 8, no. 8: 874. https://doi.org/10.3390/cells8080874