YCG063 inhibits Pseudomonas aeruginosa LPS-induced inflammation in human retinal pigment epithelial cells through the TLR2-mediated AKT/NF-κB pathway and ROS-independent pathways

Paeng,Sung Hwa; Park,Won Sun; Jung,Won-Kyo; Lee,Dae-Sung; Kim,Gi-Young; Choi,Yung Hyun; Seo,Su-Kil; Jang,Won Hee; Choi,Jung Sik; Lee,Young-Min; Park,Saegwang; Choi,Il-Whan

doi:10.3892/ijmm.2015.2266

YCG063 inhibits Pseudomonas aeruginosa LPS-induced inflammation in human retinal pigment epithelial cells through the TLR2-mediated AKT/NF-κB pathway and ROS-independent pathways

Authors:
- Sung Hwa Paeng
- Won Sun Park
- Won-Kyo Jung
- Dae-Sung Lee
- Gi-Young Kim
- Yung Hyun Choi
- Su-Kil Seo
- Won Hee Jang
- Jung Sik Choi
- Young-Min Lee
- Saegwang Park
- Il-Whan Choi
View Affiliations

Affiliations: Department of Neurosurgery, Busan Paik Hospital, Inje University College of Medicine, Busan, Republic of Korea, Department of Physiology, Kangwon National University School of Medicine, Chuncheon, Gangwon, Republic of Korea, Department of Biomedical Engineering, and Center for Marine-Integrated Biomedical Technology (BK21 Plus), Pukyong National University, Busan, Republic of Korea, Marine Biodiversity Institute of Korea, Seocheon, Chungcheongnam-do, Republic of Korea, Laboratory of Immunobiology, Department of Marine Life Sciences, Jeju National University, Jeju, Republic of Korea, Department of Biochemistry, College of Oriental Medicine, Dongeui University, Busan, Republic of Korea, Department of Microbiology, Inje University College of Medicine, Busan, Republic of Korea, Department of Biochemistry, Inje University College of Medicine, Busan, Republic of Korea, Department of Internal Medicine, Busan Paik Hospital, Inje University College of Medicine, Busan, Republic of Korea
Published online on: June 30, 2015 https://doi.org/10.3892/ijmm.2015.2266
Pages: 808-816

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

YCG063 is known as an inhibitor of reactive oxygen species (ROS); however, its intracellular mechanisms of action remain poorly understood. In the present study, we investigated the effects of YCG063 on the inflammatory response of Pseudomonas aeruginosa lipopolysaccharide (PA-LPS)‑stimulated human retinal pigment epithelial cells (RPE cells). Human adult RPE cells (ARPE‑19) were stimulated with PA-LPS. We then investigated the LPS-induced expression of several inflammatory mediators, such as interleukin (IL)-6, IL-8, monocyte chemoattractant protein-1 (MCP-1) and intracellular adhesion molecule-1 (ICAM-1) in the ARPE-19 cells. We performed an enzyme-linked immunosorbent assay (ELISA), western blot analysis, electrophoretic mobility shift assay (EMSA) and fluorescence-activated cell sorting (FACS) to elucidate the mechanisms involved in the anti-inflammatory effects of YCG063 in the PA-LPS-stimulated cells. The results revealed that treatment with YCG063 significantly inhibited the levels of IL-6, IL-8, MCP-1 and ICAM-1 in the PA-LPS-stimulated ARPE-19 cells. YCG063 also markedly inhibited the phosphorylation of AKT in the PA‑LPS-stimulated cells. In addition, the activation of nuclear factor-κB (NF-κB) was also attenuated folllowing treatment with YCG063. ROS were not generated in the PA-LPS-stimulated cells. In conclusion, our data indicate that YCG063 may prove to be a potential protective agent against inflammation, possibly through the downregulation of Toll‑like receptor 2 (TLR2) and the AKT-dependent NF-κB activation pathway in PA-LPS-stimulated ARPE-19 cells. Furthermore, this anti-inflammatory activity occurred through ROS-independent signaling pathways.

View Figures

View References

1	Bertino JS Jr: Impact of antibiotic resistance in the management of ocular infections: The role of current and future antibiotics. Clin Ophthalmol. 3:507–521. 2009. View Article : Google Scholar : PubMed/NCBI
2	Al-Amri MS: Endogenous endophthalmitis secondary to pyogenic liver abscess. Int J Diabet Mellitus. 2:646. 2010. View Article : Google Scholar
3	Callegan MC, Engelbert M, Parke DW II, Jett BD and Gilmore MS: Bacterial endophthalmitis: Epidemiology, therapeutics, and bacterium-host interactions. Clin Microbiol Rev. 15:111–124. 2002. View Article : Google Scholar : PubMed/NCBI
4	Obritsch MD, Fish DN, MacLaren R and Jung R: Nosocomial infections due to multidrug-resistant Pseudomonas aeruginosa: Epidemiology and treatment options. Pharmacotherapy. 25:1353–1364. 2005. View Article : Google Scholar : PubMed/NCBI
5	Eifrig CW, Scott IU, Flynn HW Jr and Miller D: Endophthalmitis caused by Pseudomonas aeruginosa. Ophthalmology. 110:1714–1717. 2003. View Article : Google Scholar : PubMed/NCBI
6	Ren Y, Xie Y, Jiang G, Fan J, Yeung J, Li W, Tam PK and Savill J: Apoptotic cells protect mice against lipopolysaccharide-induced shock. J Immunol. 180:4978–4985. 2008. View Article : Google Scholar : PubMed/NCBI
7	Vallejo-Garcia JL, Asencio-Duran M, Pastora-Salvador N, Vinciguerra P and Romano MR: Role of inflammation in endo-phthalmitis. Mediators Inflamm. 2012:1960942012. View Article : Google Scholar
8	Campochiaro PA: Ocular neovascularization. J Mol Med Berl. 91:311–321. 2013. View Article : Google Scholar : PubMed/NCBI
9	Bonilha VL: Age and disease-related structural changes in the retinal pigment epithelium. Clin Ophthalmol. 2:413–424. 2008. View Article : Google Scholar : PubMed/NCBI
10	Arranz-Valsero I, Schulze U, Contreras-Ruiz L, García-Posadas L, López-García A, Paulsen F and Diebold Y: Involvement of corneal epithelial cells in the Th17 response in an in vitro bacterial inflammation model. Mol Vis. 19:85–99. 2013.PubMed/NCBI
11	Gallucci RM, Simeonova PP, Matheson JM, Kommineni C, Guriel JL, Sugawara T and Luster MI: Impaired cutaneous wound healing in interleukin-6-deficient and immunosuppressed mice. FASEB J. 14:2525–2531. 2000. View Article : Google Scholar : PubMed/NCBI
12	Gallucci RM, Sugawara T, Yucesoy B, Berryann K, Simeonova PP, Matheson JM and Luster MI: Interleukin-6 treatment augments cutaneous wound healing in immunosup-pressed mice. J Interferon Cytokine Res. 21:603–609. 2001. View Article : Google Scholar : PubMed/NCBI
13	Biswas PS, Banerjee K, Kinchington PR and Rouse BT: Involvement of IL-6 in the paracrine production of VEGF in ocular HSV-1 infection. Exp Eye Res. 82:46–54. 2006. View Article : Google Scholar
14	De Vos AF, Hoekzema R and Kijlstra A: Cytokines and uveitis, a review. Curr Eye Res. 11:581–597. 1992. View Article : Google Scholar : PubMed/NCBI
15	Fenton RR, Molesworth-Kenyon S, Oakes JE and Lausch RN: Linkage of IL-6 with neutrophil chemoattractant expression in virus-induced ocular inflammation. Invest Ophthalmol Vis Sci. 43:737–743. 2002.PubMed/NCBI
16	Cole N, Bao S, Willcox M and Husband AJ: Expression of interleukin-6 in the cornea in response to infection with different strains of Pseudomonas aeruginosa. Infect Immun. 67:2497–2502. 1999.PubMed/NCBI
17	Youker K, Smith CW, Anderson DC, Miller D, Michael LH, Rossen RD and Entman ML: Neutrophil adherence to isolated adult cardiac myocytes. Induction by cardiac lymph collected during ischemia and reperfusion. J Clin Invest. 89:602–609. 1992. View Article : Google Scholar : PubMed/NCBI
18	Liu Y, Shaw SK, Ma S, Yang L, Luscinskas FW and Parkos CA: Regulation of leukocyte transmigration: Cell surface interactions and signaling events. J Immunol. 172:7–13. 2004. View Article : Google Scholar
19	Tacke F and Randolph GJ: Migratory fate and differentiation of blood monocyte subsets. Immunobiology. 211:609–618. 2006. View Article : Google Scholar : PubMed/NCBI
20	Van Riper G, Siciliano S, Fischer PA, Meurer R, Springer MS and Rosen H: Characterization and species distribution of high affinity GTP-coupled receptors for human rantes and monocyte chemoattractant protein 1. J Exp Med. 177:851–856. 1993. View Article : Google Scholar : PubMed/NCBI
21	Carr MW, Roth SJ, Luther E, Rose SS and Springer TA: Monocyte chemoattractant protein 1 acts as a T-lymphocyte chemoattractant. Proc Natl Acad Sci USA. 91:3652–3656. 1994. View Article : Google Scholar : PubMed/NCBI
22	Kawashima A, Suzuki T, Nishihara F, Kobayashi T, Takaku Y, Nakagome K, Soma T, Hagiwara K, Kanazawa M and Nagata M: Effect of formoterol on eosinophil trans-basement membrane migration induced by interleukin-8-stimulated neutrophils. Int Arch Allergy Immunol. 161(Suppl 2): 10–15. 2013. View Article : Google Scholar : PubMed/NCBI
23	Chen YJ, Huang YS, Chen JT, Chen YH, Tai MC, Chen CL and Liang CM: Protective effects of glucosamine on oxidative-stress and ischemia/reperfusion-induced retinal injury. Invest Ophthalmol Vis Sci. 56:1506–1516. 2015. View Article : Google Scholar : PubMed/NCBI
24	Kim KH, Park JY, Jung HJ and Kwon HJ: Identification and biological activities of a new antiangiogenic small molecule that suppresses mitochondrial reactive oxygen species. Biochem Biophys Res Commun. 404:541–545. 2011. View Article : Google Scholar
25	Meng Z, Yan C, Deng Q, Gao DF and Niu XL: Curcumin inhibits LPS-induced inflammation in rat vascular smooth muscle cells in vitro via ROS-relative TLR4-MAPK/NF-κB pathways. Acta Pharmacol Sin. 34:901–911. 2013. View Article : Google Scholar : PubMed/NCBI
26	Novosad BD, Astley RA and Callegan MC: Role of Toll-like receptor (TLR) 2 in experimental Bacillus cereus endo-phthalmitis. PLoS One. 6:e286192011. View Article : Google Scholar
27	Schwarz JM and Bilbo SD: LPS elicits a much larger and broader inflammatory response than Escherichia coli infection within the hippocampus of neonatal rats. Neurosci Lett. 497:110–115. 2011. View Article : Google Scholar : PubMed/NCBI
28	Aderem A and Ulevitch RJ: Toll-like receptors in the induction of the innate immune response. Nature. 406:782–787. 2000. View Article : Google Scholar : PubMed/NCBI
29	Akira S and Takeda K: Toll-like receptor signalling. Nat Rev Immunol. 4:499–511. 2004. View Article : Google Scholar : PubMed/NCBI
30	Vogel SN, Fitzgerald KA and Fenton MJ: TLRs: Differential adapter utilization by toll-like receptors mediates TLR-specific patterns of gene expression. Mol Interv. 3:466–477. 2003. View Article : Google Scholar
31	McGuire VA, Gray A, Monk CE, Santos SG, Lee K, Aubareda A, Crowe J, Ronkina N, Schwermann J, Batty IH, et al: Cross talk between the Akt and p38α pathways in macrophages downstream of Toll-like receptor signaling. Mol Cell Biol. 33:4152–4165. 2013. View Article : Google Scholar : PubMed/NCBI
32	Jung WK, Lee DY, Park C, et al: Cilostazol is anti-inflammatory in BV2 microglial cells by inactivating nuclear factor-kappaB and inhibiting mitogen-activated protein kinases. Br J Pharmacol. 159:1274–1285. 2010. View Article : Google Scholar : PubMed/NCBI
33	Dąbek J, Kułach A and Gąsior Z: Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB): a new potential therapeutic target in atherosclerosis? Pharmacol Rep. 62:778–783. 2010. View Article : Google Scholar
34	Chen BC, Wu WT, Ho FM and Lin WW: Inhibition of interleukin-1beta-induced NF-kappa B activation by calcium/calmodulin-dependent protein kinase kinase occurs through Akt activation associated with interleukin-1 receptor-associated kinase phosphorylation and uncoupling of MyD88. J Biol Chem. 277:24169–24179. 2002. View Article : Google Scholar : PubMed/NCBI
35	Li W, Yan F, Zhou H, Lin X, Wu Y, Chen C, Zhou N, Chen Z, Li JD and Shen H: P. aeruginosa lipopolysaccharide-induced MUC5AC and CLCA3 expression is partly through Duox1 in vitro and in vivo. PLoS One. 8:e639452013. View Article : Google Scholar : PubMed/NCBI