16α, 17α-epoxypregnenolone-20-oxime inhibits NO and IL-6 production in LPS-treated RAW264.7 cells

Sun,Hu‑Nan; Han,Ying‑Hao; Feng,Li; Jin,Cheng‑Hao; Han,Bing; Liu,Lei; Lee,Dong‑Soek; Kwon,Tea‑Ho; Li,Le‑Gong; Ge,Wen‑Zhong; Cui,Yu‑Dong

doi:10.3892/mmr.2016.5125

16α, 17α-epoxypregnenolone-20-oxime inhibits NO and IL-6 production in LPS-treated RAW264.7 cells

Authors:
- Hu‑Nan Sun
- Ying‑Hao Han
- Li Feng
- Cheng‑Hao Jin
- Bing Han
- Lei Liu
- Dong‑Soek Lee
- Tea‑Ho Kwon
- Le‑Gong Li
- Wen‑Zhong Ge
- Yu‑Dong Cui
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Affiliations: Laboratory of Disease Animal Model Research Center (DMARC), College of Life Science and Technology, Heilongjiang Bayi Agricultural University, Daqing, Heilongjiang 163319, P.R. China, Laboratory of Neurobiology School of Life Sciences, KNU Creative Bioresearch Group (BK21 plus project), Kyungpook National University, Daegu 702‑701, Republic of Korea, Department of Bioscience and Biotechnology, Bio/Molecular Informatics Center, Konkuk University, Seoul 143‑701, Republic of Korea
Published online on: April 14, 2016 https://doi.org/10.3892/mmr.2016.5125
Pages: 4927-4933

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Abstract

It has previously been reported that 16α, 17α-epoxypregnenolone-20-oxime (EPREGO) exerts an inhibitory effect on nitric oxide (NO) production and inducible NO synthase (iNOS) expression in microglia. The present study aimed to investigate the effects of EPREGO on the lipopolysaccharide (LPS)‑induced inflammatory response in RAW264.7 macrophage cells, and to determine the underlying molecular mechanisms using western blot analysis, enzyme‑linked immunosorbent assays and fluorescence‑activated cell sorting. The present study demonstrated that LPS‑induced production of NO and interleukin (IL)-6, and the protein expression levels of iNOS, were reduced by EPREGO in a dose‑ and time‑dependent manner, whereas, EPREGO did not affect tumor necrosis factor‑α production. In addition, EPREGO suppressed LPS‑induced cellular reactive oxygen species production and phagocytosis. Furthermore, EPREGO significantly inhibited the LPS‑induced activation of mitogen‑activated protein kinases and inhibitor of κB α degradation in LPS‑stimulated RAW264.7 cells, thus resulting in modulation of the production of NO and IL‑6. Taken together, these results suggest that EPREGO exhibits anti-inflammatory activity in macrophages, thus validating the hypothesis that EPREGO may be useful as a therapeutic agent for the treatment of macrophage-mediated inflammation.

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1	Underhill DM and Ozinsky A: Phagocytosis of microbes: Complexity in action. Annu Rev Immunol. 20:825–852. 2002. View Article : Google Scholar : PubMed/NCBI
2	Li W, Ashok M, Li J, Yang H, Sama AE and Wang H: A major ingredient of green tea rescues mice from lethal sepsis partly by inhibiting HMGB1. PLoS One. 2:e11532007. View Article : Google Scholar : PubMed/NCBI
3	Takeuchi O and Akira S: Pattern recognition receptors and inflammation. Cell. 140:805–820. 2010. View Article : Google Scholar : PubMed/NCBI
4	Cohen J: The immunopathogenesis of sepsis. Nature. 420:885–891. 2002. View Article : Google Scholar : PubMed/NCBI
5	Guha M and Mackman N: LPS induction of gene expression in human monocytes. Cell Signal. 13:85–94. 2001. View Article : Google Scholar : PubMed/NCBI
6	Ehlting C, Ronkina N, Böhmer O, Albrecht U, Bode KA, Lang KS, Kotlyarov A, Radzioch D, Gaestel M, Häussinger D and Bode JG: Distinct functions of the mitogen-activated protein kinase-activated protein (MAPKAP) kinases MK2 and MK3: MK2 mediates lipopolysaccharide-induced signal transducers and activators of transcription 3 (STAT3) activation by preventing negative regulatory effects of MK3. J Biol Chem. 286:24113–24124. 2011. View Article : Google Scholar : PubMed/NCBI
7	Hsieh TP, Sheu SY, Sun JS and Chen MH: Icariin inhibits osteoclast differentiation and bone resorption by suppression of MAPKs/NF-κB regulated HIF-1α and PGE(2) synthesis. Phytomedicine. 18:176–185. 2011. View Article : Google Scholar
8	Shin JS, Noh YS, Lee YS, Cho YW, Baek NI, Choi MS, Jeong TS, Kang E, Chung HG and Lee KT: Arvelexin from Brassica rapa suppresses NF-κB-regulated pro-inflammatory gene expression by inhibiting activation of IκB kinase. Br J Pharmacol. 164:145–158. 2011. View Article : Google Scholar : PubMed/NCBI
9	Kim JH, Kim DH, Baek SH, Lee HJ, Kim MR, Kwon HJ and Lee CH: Rengyolone inhibits inducible nitric oxide synthase expression and nitric oxide production by down-regulation of NF-kappaB and p38 MAP kinase activity in LPS-stimulated RAW 264.7 cells. Biochem Pharmacol. 71:1198–1205. 2006. View Article : Google Scholar : PubMed/NCBI
10	Al-Mutairi MS, Cadalbert LC, McGachy HA, Shweash M, Schroeder J, Kurnik M, Sloss CM, Bryant CE, Alexander J and Plevin R: MAP kinase phosphatase-2 plays a critical role in response to infection by Leishmania mexicana. PLoS Pathog. 6:e10011922010. View Article : Google Scholar : PubMed/NCBI
11	Liu HE, Chang AS, Teng CM, Chen CC, Tsai AC and Yang CR: Potent anti-inflammatory effects of denbinobin mediated by dual inhibition of expression of inducible no synthase and cyclooxygenase 2. Shock. 35:191–197. 2011. View Article : Google Scholar
12	Liou HC: Regulation of the immune system by NF-kappaB and IkappaB. J Biochem Mol Biol. 35:537–546. 2002. View Article : Google Scholar : PubMed/NCBI
13	Karin M and Delhase M: The I kappaB kinase (IKK) and NF-kappa B: Key elements of proinflammatory signalling. Semin Immunol. 12:85–98. 2000. View Article : Google Scholar : PubMed/NCBI
14	Baeuerle PA and Baltimore D: NF-kappaB: Ten years after. Cell. 87:13–20. 1996. View Article : Google Scholar : PubMed/NCBI
15	Sun HN, Jin MH, Han B, Feng L, Han YH, Shen GN, Yu YZ, Jin CH, Lian ZX, Lee DS, et al: 16α, 17α-Epoxypregnenolone-20-oxime prevent LPS-induced NO production and iNOS expression in BV-2 microglial cells by inhibiting JNK phosphorylation. Biol Pharm Bull. 37:1096–1102. 2014. View Article : Google Scholar
16	Liu Y, Hao W, Letiembre M, Walter S, Kulanga M, Neumann H and Fassbender K: Suppression of microglial inflammatory activity by myelin phagocytosis: Role of p47-PHOX-mediated generation of reactive oxygen species. J Neurosci. 26:12904–12913. 2006. View Article : Google Scholar : PubMed/NCBI
17	Ritchlin CT, Haas-Smith SA, Li P, Hicks DG and Schwarz EM: Mechanisms of TNF-alpha- and RANKL-mediated osteoclastogenesis and bone resorption in psoriatic arthritis. J Clin Invest. 111:821–831. 2003. View Article : Google Scholar : PubMed/NCBI
18	Chowdhury P, Das AM and Goswami P: Synthesis of some new steroidal [16alpha, 17alpha-d]-isoxazolines. Steroids. 70:494–498. 2005. View Article : Google Scholar : PubMed/NCBI
19	Banerjee T, Shukla A, Shinde K and Patil S: Mixed culture bioconversion of 16-dehydropregnenolone acetate to androsta-1,4-diene-3,17-dione: Optimization of parameters. Biotechnol Prog. 19:662–664. 2003. View Article : Google Scholar : PubMed/NCBI
20	Lu YC, Yeh WC and Ohashi PS: LPS/TLR4 signal transduction pathway. Cytokine. 42:145–151. 2008. View Article : Google Scholar : PubMed/NCBI
21	Pålsson-McDermott EM and O'Neill LA: Signal transduction by the lipopolysaccharide receptor, Toll-like receptor-4. Immunology. 113:153–162. 2004. View Article : Google Scholar : PubMed/NCBI
22	Cao W, Bao C, Padalko E and Lowenstein CJ: Acetylation of mitogen-activated protein kinase phosphatase-1 inhibits Toll-like receptor signaling. J Exp Med. 205:1491–1503. 2008. View Article : Google Scholar : PubMed/NCBI
23	Cheng YW, Chang CY, Lin KL, Hu CM, Lin CH and Kang JJ: Shikonin derivatives inhibited LPS-induced NOS in RAW 264.7 cells via downregulation of MAPK/NF-kappaB signaling. J Ethnopharmacol. 120:264–271. 2008. View Article : Google Scholar : PubMed/NCBI
24	Xie C, Kang J, Ferguson ME, Nagarajan S, Badger TM and Wu X: Blueberries reduce pro-inflammatory cytokine TNF-α and IL-6 production in mouse macrophages by inhibiting NF-κB activation and the MAPK pathway. Mol Nutr Food Res. 55:1587–1591. 2011. View Article : Google Scholar : PubMed/NCBI
25	Chu X, Ci X, He J, Wei M, Yang X, Cao Q, Li H, Guan S, Deng Y, Pang D and Deng X: A novel anti-inflammatory role for ginkgolide B in asthma via inhibition of the ERK/MAPK signaling pathway. Molecules. 16:7634–7648. 2011. View Article : Google Scholar : PubMed/NCBI
26	Kim HG, Kim NR, Gim MG, Lee JM, Lee SY, Ko MY, Kim JY, Han SH and Chung DK: Lipoteichoic acid isolated from Lactobacillus plantarum inhibits lipopolysaccharide-induced TNF-alpha production in THP-1 cells and endotoxin shock in mice. J Immunol. 180:2553–2561. 2008. View Article : Google Scholar : PubMed/NCBI
27	Jin S, Park JY, Hong JM, Kim TH, Shin HI, Park EK and Kim SY: Inhibitory effect of (−)-epigallocatechin gallate on titanium particle-induced TNF-α release and in vivo osteolysis. Exp Mol Med. 43:411–418. 2011. View Article : Google Scholar : PubMed/NCBI
28	Li M, Zhong X, He Z, Wen M, Li J, Peng X, Liu G, Deng J, Zhang J and Bai J: Effect of erythromycin on cigarette-induced histone deacetylase protein expression and nuclear factor-κB activity in human macrophages in vitro. Int Immunopharmacol. 12:643–650. 2012. View Article : Google Scholar : PubMed/NCBI
29	Li YQ, Zhang ZX, Xu YJ, Ni W, Chen SX, Yang Z and Ma D: N-Acetyl-L-cysteine and pyrrolidine dithiocarbamate inhibited nuclear factor-kappaB activation in alveolar macrophages by different mechanisms. Acta Pharmacol Sin. 27:339–346. 2006. View Article : Google Scholar : PubMed/NCBI
30	Korobowicz A: Biology of tumor necrosis factor type alpha (TNF-alpha). Pol Merkur Lekarski. 21:358–361. 2006.In Polish.
31	Shuai K and Liu B: Regulation of JAK-STAT signalling in the immune system. Nat Rev Immunol. 3:900–911. 2003. View Article : Google Scholar : PubMed/NCBI
32	Ohmori Y and Hamilton TA: Requirement for STAT1 in LPS-induced gene expression in macrophages. J Leukoc Biol. 69:598–604. 2001.PubMed/NCBI
33	Simon AR, Rai U, Fanburg BL and Cochran BH: Activation of the JAK-STAT pathway by reactive oxygen species. Am J Physiol. 275:C1640–C1652. 1998.PubMed/NCBI