Wonmin Ko; Jae Hak Sohn; Youn-Chul Kim; Hyuncheol Oh

doi:10.20307/nps.2015.21.4.240

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Ko, Sohn, Kim, and Oh: Viridicatol from Marine-derived Fungal Strain Penicillium sp. SF-5295 Exerts Anti-inflammatory Effects through Inhibiting NF-κ B Signaling Pathway on Lipopolysaccharide-induced RAW264.7 and BV2 Cells

Original Article

Natural Product Sciences 2015;21(4):240-247.

Published online: 17 January 2015

DOI: https://doi.org/10.20307/nps.2015.21.4.240

Viridicatol from Marine-derived Fungal Strain Penicillium sp. SF-5295 Exerts Anti-inflammatory Effects through Inhibiting NF-κ B Signaling Pathway on Lipopolysaccharide-induced RAW264.7 and BV2 Cells

Wonmin Ko¹, Jae Hak Sohn², Youn-Chul Kim^1,^∗, Hyuncheol Oh^1,^∗

¹ College of Pharmacy, Wonkwang University, Iksan 570-749, Korea

² College of Medical and Life Sciences, Silla University, Busan 617-736, Republic of Korea

^∗Author for correspondence Youn-Chul Kim, College of Pharmacy, Wonkwang University, Iksan 570-749, Republic of Korea Tel: +82-63-850-6823; E-mail: yckim@wku.ac.kr

Hyuncheol Oh, College of Pharmacy, Wonkwang University, Iksan 570-749, Republic of Korea Tel: +82-63-850-6815; E-mail: hoh@wku.ac.kr

Revised 29 June 20154 August 2015 Accepted 5 August 2015

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Viridicatol (1) has previously been isolated from the extract of the marine-derived fungus Penicillium sp. SF-5295. In the course of further biological evaluation of this quinolone alkaloid, anti-inflammatory effect of 1 in RAW264.7 and BV2 cells stimulated with lipopolysaccharide (LPS) was observed. In this study, our data indicated that 1 suppressed the expression of well-known proinflammatory mediators such as inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2, and consequently inhibited the production of iNOS-derived nitric oxide (NO) and COX-2-derived prostaglandin E2 (PGE₂) in LPS stimulated RAW264.7 and BV2 cells. Compound 1 also reduced mRNA expression of proinflammatory cytokines such as interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α). In the further evaluation of the mechanisms of these anti-inflammatory effects, 1 was shown to inhibit nuclear factor-kappa B (NF-κB) pathway in LPS-stimulated RAW264.7 and BV2 cells. Compound 1 blocked the phosphorylation and degradation of inhibitor kappa B (IκB)-α in the cytoplasm, and suppressed the translocation of NF-κ B p65 and p50 heterodimer in nucleus. In addition, viridicatol (1) attenuated the DNA-binding activity of NF-κ B in LPS-stimulated RAW264.7 and BV2 cells.

Keywords: Viridicatol, Penicillium sp., Marine fungus, Anti-neuroinflammation, Nuclear factor-kappa B (NF-κ B)

REFERENCES

(1). Ingersoll M. A., Platt A. M., Potteaux S., Randolph G. J.Trends Immunol. 2011; 32:470–477.

(2). Sosroseno W., Barid I., Herminajeng E., Susilowati H.Oral Microbiol. Immunol. 2002; 17:72–78.

(3). Block M. L., Zecca L., Hong J. S.Nat. Rev. Neurosci. 2007; 8:57–69.

(4). Jiang F., Ramanathan A., Miller M. T., Tang G. Q., Gale M. Jr., Patel S. S., Marcotrigiano J.Nature. 2011; 479:423–427.

(5). Tsan M. F., Gao B. J.Endotoxin Res. 2007; 13:6–14.

(6). Yang Y. Z., Tang Y. Z., Liu Y. H. J.Ethnopharmacol. 2013; 148:271–276.

(7). Mizuno T., Kurotani T., Komatsu Y., Kawanokuchi J., Kato H., Mitsuma N., Suzumura A.Neuropharmacology. 2004; 46:404–411.

(8). Ivashkiv L. B.Eur. J. Immunol. 2011; 41:2477–2481.

(9). Baldwin A. S.Jr. Annu. Rev. Immunol. 1996; 14:649–683.

(10). Mancino A., Lawrence T.Clin. Cancer Res. 2010; 16:784–789.

(11). Bremner P., Heinrich M. J.Pharm. Pharmacol. 2002; 54:453–472.

(12). Rateb M. E., Ebel R.Nat. Prod. Rep. 2011; 28:290–334.

(13). Bugni T. S., Ireland C. M.Nat. Prod. Rep. 2004; 21:143–163.

(14). Fremlin L. J., Piggott A. M., Lacey E., Capon R. J. J.Nat. Prod. 2009; 72:666–670.

(15). Sohn J. H., Lee Y. R., Lee D. S., Kim Y. C., Oh H. J.Microbiol. Biotechnol. 2013; 23:1206–1211.

(16). Berridge M. V., Tan A. S.Arch. Biochem. Biophys. 1993; 303:474–482.

(17). Titheradge M. A.Methods Mol. Biol. 1998; 100:83–91.

(18). Karin M., Ben-Neriah Y.Annu. Rev. Immunol. 2000; 18:621–663.

(19). Grilli M., Chiu J. J., Lenardo M. J.Int. Rev. Cytol. 1993; 143:1–62.

(20). Chen F., Sun S. C., Kuh D. C., Gaydos L. J., Demers L. M.Biochem. Biophys. Res. Commun. 1995; 214:985–992.

(21). Saleem M., Ali M. S., Hussain S., Jabbar A., Ashraf M., Lee Y. S.Nat. Prod. Rep. 2007; 24:1142–1152.

(22). Fenical W., Jensen P. R.Nat. Chem. Biol. 2006; 2:666–673.

(23). Lee D. S., Ko W., Quang T. H., Kim K. S., Sohn J. H., Jang J. H., Ahn J. S., Kim Y. C., Oh H.Mar. Drugs. 2013; 11:4510–4526.

(24). Su X., Chen Q., Chen W., Chen T., Li W., Li Y., Dou X., Zhang Y., Shen Y., Wu H., Yu C.Int. Immunopharmacol. 2014; 19:88–93.

(25). Wei M. Y., Yang R. Y., Shao C. L., Wang C. Y., Deng D. S., She Z. G., Lin Y. C.Chem. Nat. Compd. 2011; 47:322–325.

Fig. 1.

Chemical structure of viridicatol (1).

Fig. 2.

Effects of 1 on cell viability. RAW264.7 (A) and BV2 cells (B) were incubated for 24 h with various concentrations of 1 (1 - 160μ M). Cell viability was determined by MTT assay. Each bar represents the mean ± S.D. of three independent experiments.

Fig. 3.

Effects of 1 on the protein expression of nitrite (A), PGE₂ (B) iNOS (C) and COX-2 (D) in RAW 264.7 cells stimulated with LPS. The cells were pre-treated for 3 h with indicated concentrations of 1, and then stimulated for 24 h with LPS (1 μ g/mL). The concentrations of nitrite, PGE₂ and Western blot analysis were performed as described in the experimental section, and representative data of three independent experiments are shown. ^∗p < 0.05 compared with the LPS.

Fig. 4.

Effects of 1 on the protein expression of nitrite (A), PGE₂ (B) iNOS (C) and COX-2 (D) in BV2 cells stimulated with LPS. The cells were pre-treated for 3 h with indicated concentrations of 1, and then stimulated for 24 h with LPS (1 μg/mL). The concentrations of nitrite, PGE₂ and Western blot analysis were performed as described in the experimental section, and representative data of three independent experiments are shown. ^∗p < 0.05 compared with the LPS.

Fig. 5.

Effects of 1 on IL-1β mRNA (A), IL-6 mRNA (B), and TNF-α mRNA (C) expression in RAW264.7 cells. Cells were incubated with the indicated concentrations of 1 for 3 h, and then treated for 6 h with LPS (1 μg/mL). RNA quantification for IL-1β, IL-6 and TNF-α expression were performed as described in the experimental section. Data shown represent the mean values of three independent experiments. ^∗p < 0.05 compared with the LPS.

Fig. 6.

Effects of 1 on IL-1β mRNA (A), IL-6 mRNA (B), and TNF-α mRNA (C) expression in BV2 cells. Cells were incubated with the indicated concentrations of 1 for 3 h, and then treated for 6 h with LPS (1 μg/mL). RNA quantification for IL-1β, IL-6 and TNF-α expression were performed, as described in the experimental section. Data shown represent the mean values of three independent experiments. ^∗p < 0.05 compared with the LPS.

Fig. 7.

Effects of 1 on the Iκ B-α phosphorylation and degradation (A), NF-κ B activation (p65 and p50), and NF-κ B DNA-binding activity (C). RAW264.7 cells were pre-treated with the indicated concentrations of 1 for 3 h, and then stimulated with LPS (1 μg/mL) for 1 h. The western blot analysis of IκBα and p-IκBα in the cytoplasm and NF-κB in the nucleus was performed as described in the experimental section. The representative blots of three independent experiments are shown. A commercially available NF-κB ELISA (Active Motif) was then used to test nuclear extracts and to determine the degree of NF-κB binding. The data represent the mean values of 3 experiments ± SD. ^∗P < 0.05 compared with the group treated with LPS.

Fig. 8.

Effects of 1 on the Iκ B-α phosphorylation and degradation (A), NF-κ B activation (p65 and p50), and NF-κ B DNA-binding activity (C). BV2 cells were pre-treated with the indicated concentrations of 1 for 3 h, and then stimulated with LPS (1 μg/mL) for 1 h. The western blot analysis of Iκ Bα and p-Iκ Bα in the cytoplasm and NF-κ B in the nucleus was performed as described in the experimental section. The representative blots of three independent experiments are shown. A commercially available NF-κB ELISA (Active Motif) was then used to test nuclear extracts and to determine the degree of NF-κB binding. The data represent the mean values of 3 experiments ± SD. ^∗P < 0.05 compared with the group treated with LPS.

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