Anti-Inflammatory Property of 5-Demethylnobiletin (5-Hydroxy-6, 7, 8, 3′, 4′-pentamethoxyflavone) and Its Metabolites in Lipopolysaccharide (LPS)-Induced RAW 264.7 Cells
Abstract
Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Chemicals
2.2. Cell Culture
2.3. Cell Viability Assay
2.4. Nitric Oxide Assay
2.5. ELISA for Interleukin-1β (IL-1β)
2.6. Preparation of Whole-Cell Lysate
2.7. Immunoblot Analysis
2.8. Quantitative Real-Time Reverse-Transcription Polymerase Chain Reaction (qRT-PCR) Analysis
2.9. Statistical Analysis
3. Results
3.1. 5DN and Its Metabolites (M1, M2, and M3) Inhibit NO Production in LPS-Induced RAW 264.7 Cells
3.2. 5DN and Its Metabolites (M2 and M3) Suppress iNOS and COX-2 Gene Expression in LPS-Induced RAW 264.7 Cells
3.3. 5DN and Its Metabolites (M2 and M3) Attenuate IL-1β Gene Expression in LPS-Induced RAW 264.7 Cells
3.4. 5DN and Its Metabolites (M1 and M3) Increase HO-1 Gene Expression in LPS-Induced RAW 264.7 Cells
4. Discussion
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Coussens, L.M.; Werb, Z. Inflammation and cancer. Nature 2002, 420, 860–867. [Google Scholar] [CrossRef] [PubMed]
- Li, S.; Lo, C.Y.; Ho, C.T. Hydroxylated polymethoxyflavones and methylated flavonoids in sweet orange (Citrus sinensis) peel. J. Agric. Food Chem. 2006, 54, 4176–4185. [Google Scholar] [CrossRef] [PubMed]
- Murakami, A.; Nakamura, Y.; Ohto, Y.; Yano, M.; Koshiba, T.; Koshimizu, K.; Tokuda, H.; Nishino, H.; Ohigashi, H. Suppressive effects of citrus fruits on free radical generation and nobiletin, an anti-inflammatory polymethoxyflavonoid. Biofactors 2000, 12, 187–192. [Google Scholar] [CrossRef] [PubMed]
- Murakami, A.; Shigemori, T.; Ohigashi, H. Zingiberaceous and citrus constituents, 1’-acetoxychavicol acetate, zerumbone, auraptene, and nobiletin, suppress lipopolysaccharide-induced cyclooxygenase-2 expression in RAW264.7 murine macrophages through different modes of action. J. Nutr. 2005, 135, 2987S–2992S. [Google Scholar] [CrossRef]
- Lai, C.S.; Li, S.; Chai, C.Y.; Lo, C.Y.; Ho, C.T.; Wang, Y.J.; Pan, M.H. Inhibitory effect of citrus 5-hydroxy-3,6,7,8,3’,4’-hexamethoxyflavone on 12-O-tetradecanoylphorbol 13-acetate-induced skin inflammation and tumor promotion in mice. Carcinogenesis 2007, 28, 2581–2588. [Google Scholar] [CrossRef]
- Murakami, A.; Nakamura, Y.; Torikai, K.; Tanaka, T.; Koshiba, T.; Koshimizu, K.; Kuwahara, S.; Takahashi, Y.; Ogawa, K.; Yano, M.; et al. Inhibitory effect of citrus nobiletin on phorbol ester-induced skin inflammation, oxidative stress, and tumor promotion in mice. Cancer Res. 2000, 60, 5059–5066. [Google Scholar]
- Tang, M.; Ogawa, K.; Asamoto, M.; Hokaiwado, N.; Seeni, A.; Suzuki, S.; Takahashi, S.; Tanaka, T.; Ichikawa, K.; Shirai, T. Protective effects of citrus nobiletin and auraptene in transgenic rats developing adenocarcinoma of the prostate (TRAP) and human prostate carcinoma cells. Cancer Sci. 2007, 98, 471–477. [Google Scholar] [CrossRef]
- Wu, X.; Li, Z.; Sun, Y.; Li, F.; Gao, Z.; Zheng, J.; Xiao, H. Identification of Xanthomicrol as a major metabolite of 5-demethyltangeretin in mouse gastrointestinal tract and its inhibitory effects on colon cancer cells. Front. Nutr. 2020, 7, 103. [Google Scholar] [CrossRef]
- Kurowska, E.M.; Manthey, J.A. Hypolipidemic effects and absorption of citrus polymethoxylated flavones in hamsters with diet-induced hypercholesterolemia. J. Agric. Food Chem. 2004, 52, 2879–2886. [Google Scholar] [CrossRef]
- Saito, T.; Abe, D.; Sekiya, K. Nobiletin enhances differentiation and lipolysis of 3T3-L1 adipocytes. Biochem. Biophys. Res. Commun. 2007, 357, 371–376. [Google Scholar] [CrossRef]
- Li, S.; Pan, M.H.; Lai, C.S.; Lo, C.Y.; Dushenkov, S.; Ho, C.T. Isolation and syntheses of polymethoxyflavones and hydroxylated polymethoxyflavones as inhibitors of HL-60 cell lines. Bioorg. Med. Chem. 2007, 15, 3381–3389. [Google Scholar] [CrossRef] [PubMed]
- Li, S.; Lambros, T.; Wang, Z.; Goodnow, R.; Ho, C.T. Efficient and scalable method in isolation of polymethoxyflavones from orange peel extract by supercritical fluid chromatography. J. Chromatogr. B Anal. Technol. Biomed. Life Sci. 2007, 846, 291–297. [Google Scholar] [CrossRef] [PubMed]
- Ishiwa, J.; Sato, T.; Mimaki, Y.; Sashida, Y.; Yano, M.; Ito, A. A citrus flavonoid, nobiletin, suppresses production and gene expression of matrix metalloproteinase 9/gelatinase B in rabbit synovial fibroblasts. J. Rheumatol. 2000, 27, 20–25. [Google Scholar] [PubMed]
- Mora, A.; Paya, M.; Rios, J.L.; Alcaraz, M.J. Structure-activity relationships of polymethoxyflavones and other flavonoids as inhibitors of non-enzymic lipid peroxidation. Biochem. Pharmacol. 1990, 40, 793–797. [Google Scholar] [CrossRef] [PubMed]
- Huguet, A.I.; Manez, S.; Alcaraz, M.J. Superoxide scavenging properties of flavonoids in a non-enzymic system. Z. Nat. C 1990, 45, 19–24. [Google Scholar] [CrossRef] [PubMed]
- Bas, E.; Recio, M.C.; Giner, R.M.; Manez, S.; Cerda-Nicolas, M.; Rios, J.L. Anti-inflammatory activity of 5-O-demethylnobiletin, a polymethoxyflavone isolated from Sideritis tragoriganum. Planta Med. 2006, 72, 136–142. [Google Scholar] [CrossRef] [PubMed]
- Qiu, P.; Dong, P.; Guan, H.; Li, S.; Ho, C.T.; Pan, M.H.; McClements, D.J.; Xiao, H. Inhibitory effects of 5-hydroxy polymethoxyflavones on colon cancer cells. Mol. Nutr. Food Res. 2010, 54, S244–S252. [Google Scholar] [CrossRef]
- Kim, H.; Moon, J.Y.; Mosaddik, A.; Cho, S.K. Induction of apoptosis in human cervical carcinoma HeLa cells by polymethoxylated flavone-rich Citrus grandis Osbeck (Dangyuja) leaf extract. Food Chem. Toxicol. 2010, 48, 2435–2442. [Google Scholar] [CrossRef]
- Xiao, H.; Yang, C.S.; Li, S.; Jin, H.; Ho, C.T.; Patel, T. Monodemethylated polymethoxyflavones from sweet orange (Citrus sinensis) peel inhibit growth of human lung cancer cells by apoptosis. Mol. Nutr. Food Res. 2009, 53, 398–406. [Google Scholar] [CrossRef]
- Zheng, J.; Song, M.; Dong, P.; Qiu, P.; Guo, S.; Zhong, Z.; Li, S.; Ho, C.T.; Xiao, H. Identification of novel bioactive metabolites of 5-demethylnobiletin in mice. Mol. Nutr. Food Res. 2013, 57, 1999–2007. [Google Scholar] [CrossRef]
- Fushiya, S.; Kishi, Y.; Hattori, K.; Batkhuu, J.; Takano, F.; Singab, A.N.; Okuyama, T. Flavonoids from Cleome droserifolia suppress NO production in activated macrophages in vitro. Planta Med. 1999, 65, 404–407. [Google Scholar] [CrossRef] [PubMed]
- Kim, H.; Lee, H.S.; Chang, K.T.; Ko, T.H.; Baek, K.J.; Kwon, N.S. Chloromethyl ketones block induction of nitric oxide synthase in murine macrophages by preventing activation of nuclear factor-kappa B. J. Immunol. 1995, 154, 4741–4748. [Google Scholar] [PubMed]
- Guo, S.; Qiu, P.; Xu, G.; Wu, X.; Dong, P.; Yang, G.; Zheng, J.; McClements, D.J.; Xiao, H. Synergistic anti-inflammatory effects of nobiletin and sulforaphane in lipopolysaccharide-stimulated RAW 264.7 cells. J. Agric. Food Chem. 2012, 60, 2157–2164. [Google Scholar] [CrossRef] [PubMed]
- Guo, S.; Wu, X.; Zheng, J.; Charoensinphon, N.; Dong, P.; Qiu, P.; Song, M.; Tang, Z.; Xiao, H. Anti-inflammatory effect of xanthomicrol, a major colonic metabolite of 5-demethyltangeretin. Food Funct. 2018, 9, 3104–3113. [Google Scholar] [CrossRef]
- Pan, M.H.; Lai, C.S.; Wang, Y.J.; Ho, C.T. Acacetin suppressed LPS-induced up-expression of iNOS and COX-2 in murine macrophages and TPA-induced tumor promotion in mice. Biochem. Pharmacol. 2006, 72, 1293–1303. [Google Scholar] [CrossRef]
- Cheung, K.L.; Khor, T.O.; Kong, A.N. Synergistic effect of combination of phenethyl isothiocyanate and sulforaphane or curcumin and sulforaphane in the inhibition of inflammation. Pharm. Res. 2009, 26, 224–231. [Google Scholar] [CrossRef]
- Livak, K.J.; Schmittgen, T.D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 2001, 25, 402–408. [Google Scholar] [CrossRef]
- Campbell, N.K.; Fitzgerald, H.K.; Dunne, A. Regulation of inflammation by the antioxidant haem oxygenase 1. Nat. Rev. Immunol. 2021, 21, 411–425. [Google Scholar] [CrossRef]
- Li, S.; Sang, S.; Pan, M.H.; Lai, C.S.; Lo, C.Y.; Yang, C.S.; Ho, C.T. Anti-inflammatory property of the urinary metabolites of nobiletin in mouse. Bioorg. Med. Chem. Lett. 2007, 17, 5177–5181. [Google Scholar] [CrossRef]
- Hibbs, J.B., Jr. Synthesis of nitric oxide from L-arginine: A recently discovered pathway induced by cytokines with antitumour and antimicrobial activity. Res. Immunol. 1991, 142, 565–569; discussion 596–598. [Google Scholar] [CrossRef]
- Dinarello, C.A. The paradox of pro-inflammatory cytokines in cancer. Cancer Metastasis Rev. 2006, 25, 307–313. [Google Scholar] [CrossRef]
- Lin, W.W.; Karin, M. A cytokine-mediated link between innate immunity, inflammation, and cancer. J. Clin. Investig. 2007, 117, 1175–1183. [Google Scholar] [CrossRef] [PubMed]
- Chen, X.L.; Kunsch, C. Induction of cytoprotective genes through Nrf2/antioxidant response element pathway: A new therapeutic approach for the treatment of inflammatory diseases. Curr. Pharm. Des. 2004, 10, 879–891. [Google Scholar] [CrossRef] [PubMed]
- Li, N.; Alam, J.; Venkatesan, M.I.; Eiguren-Fernandez, A.; Schmitz, D.; Di Stefano, E.; Slaughter, N.; Killeen, E.; Wang, X.; Huang, A.; et al. Nrf2 is a key transcription factor that regulates antioxidant defense in macrophages and epithelial cells: Protecting against the proinflammatory and oxidizing effects of diesel exhaust chemicals. J. Immunol. 2004, 173, 3467–3481. [Google Scholar] [CrossRef] [PubMed]
- Syapin, P.J. Regulation of haeme oxygenase-1 for treatment of neuroinflammation and brain disorders. Br. J. Pharmacol. 2008, 155, 623–640. [Google Scholar] [CrossRef]
- Innamorato, N.G.; Lastres-Becker, I.; Cuadrado, A. Role of microglial redox balance in modulation of neuroinflammation. Curr. Opin. Neurol. 2009, 22, 308–314. [Google Scholar] [CrossRef]
- Jeon, W.K.; Kim, B.C. WITHDRAWN: Heme oxygenase-1 mediates the anti-inflammatory effect of propyl gallate in LPS-stimulated macrophages. Biochem. Biophys. Res. Commun. 2007, 361, 645–650. [Google Scholar] [CrossRef]
- Lee, T.S.; Chau, L.Y. Heme oxygenase-1 mediates the anti-inflammatory effect of interleukin-10 in mice. Nat. Med. 2002, 8, 240–246. [Google Scholar] [CrossRef]
- Lee, T.S.; Tsai, H.L.; Chau, L.Y. Induction of heme oxygenase-1 expression in murine macrophages is essential for the anti-inflammatory effect of low dose 15-deoxy-Delta 12,14-prostaglandin J2. J. Biol. Chem. 2003, 278, 19325–19330. [Google Scholar] [CrossRef]
- Lin, C.C.; Liu, X.M.; Peyton, K.; Wang, H.; Yang, W.C.; Lin, S.J.; Durante, W. Far infrared therapy inhibits vascular endothelial inflammation via the induction of heme oxygenase-1. Arter. Thromb Vasc. Biol. 2008, 28, 739–745. [Google Scholar] [CrossRef]
- Wang, W.W.; Smith, D.L.; Zucker, S.D. Bilirubin inhibits iNOS expression and NO production in response to endotoxin in rats. Hepatology 2004, 40, 424–433. [Google Scholar] [CrossRef] [PubMed]
- Ryter, S.W.; Alam, J.; Choi, A.M. Heme oxygenase-1/carbon monoxide: From basic science to therapeutic applications. Physiol. Rev. 2006, 86, 583–650. [Google Scholar] [CrossRef] [PubMed]
- Thorup, C.; Jones, C.L.; Gross, S.S.; Moore, L.C.; Goligorsky, M.S. Carbon monoxide induces vasodilation and nitric oxide release but suppresses endothelial NOS. Am. J. Physiol. 1999, 277, F882–F889. [Google Scholar] [CrossRef] [PubMed]
- Wink, M. Evolutionary advantage and molecular modes of action of multi-component mixtures used in phytomedicine. Curr. Drug Metab. 2008, 9, 996–1009. [Google Scholar] [CrossRef] [PubMed]
- Liang, Y.C.; Huang, Y.T.; Tsai, S.H.; Lin-Shiau, S.Y.; Chen, C.F.; Lin, J.K. Suppression of inducible cyclooxygenase and inducible nitric oxide synthase by apigenin and related flavonoids in mouse macrophages. Carcinogenesis 1999, 20, 1945–1952. [Google Scholar] [CrossRef]
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Guo, S.; Wu, X.; Zheng, J.; Song, M.; Dong, P.; Xiao, H. Anti-Inflammatory Property of 5-Demethylnobiletin (5-Hydroxy-6, 7, 8, 3′, 4′-pentamethoxyflavone) and Its Metabolites in Lipopolysaccharide (LPS)-Induced RAW 264.7 Cells. Biology 2022, 11, 1820. https://doi.org/10.3390/biology11121820
Guo S, Wu X, Zheng J, Song M, Dong P, Xiao H. Anti-Inflammatory Property of 5-Demethylnobiletin (5-Hydroxy-6, 7, 8, 3′, 4′-pentamethoxyflavone) and Its Metabolites in Lipopolysaccharide (LPS)-Induced RAW 264.7 Cells. Biology. 2022; 11(12):1820. https://doi.org/10.3390/biology11121820
Chicago/Turabian StyleGuo, Shanshan, Xian Wu, Jinkai Zheng, Mingyue Song, Ping Dong, and Hang Xiao. 2022. "Anti-Inflammatory Property of 5-Demethylnobiletin (5-Hydroxy-6, 7, 8, 3′, 4′-pentamethoxyflavone) and Its Metabolites in Lipopolysaccharide (LPS)-Induced RAW 264.7 Cells" Biology 11, no. 12: 1820. https://doi.org/10.3390/biology11121820
APA StyleGuo, S., Wu, X., Zheng, J., Song, M., Dong, P., & Xiao, H. (2022). Anti-Inflammatory Property of 5-Demethylnobiletin (5-Hydroxy-6, 7, 8, 3′, 4′-pentamethoxyflavone) and Its Metabolites in Lipopolysaccharide (LPS)-Induced RAW 264.7 Cells. Biology, 11(12), 1820. https://doi.org/10.3390/biology11121820