The peptide lycosin-I attenuates TNF-α-induced inflammation in human umbilical vein endothelial cells via IκB/NF-κB signaling pathway
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The peptide lycosin-I has anti-bacterial and anti-cancer capacities. However, the anti-inflammatory activity of lycosin-I remains unknown. We investigated whether lycosin-I could attenuate inflammation.
Materials and methods
Human umbilical vein endothelial cells (HUVECs) were treated with lycosin-I before exposure to tumor necrosis factor-α (TNF-α). The expression of intercellular cell adhesion molecule-1 (ICAM-1), nuclear transcription factor-kappa B (NF-κB) p65 and inhibitory subunit of NF-κB alpha (IκBα) was evaluated by western blot. The expression of interleukin-6 (IL-6) and interleukin-8 (IL-8) was detected by quantitative RT-PCR or ELISA. Immunofluorescence analysis was used to determine the impact of lycosin-I on NF-κB pathway. C57BL/6 mice were pretreated with lycosin-I before exposure with lipopolysaccharide (LPS).
Lycosin-I significantly reduced the TNF-α-enhanced expression of IL-6, IL-8 and ICAM-1. Lycosin-I also inhibited the human monocyte cells adhesion to HUVECs. We further demonstrated that lycosin-I could effectively suppress the reaction of endothelial cells to TNF-α by inhibiting IκBα degradation. Subsequently, the phosphorylation and translocation of NF-κB p65 could also be attenuated. Furthermore, lycosin-I exhibited a significant protection of C57BL/6 mice against LPS-induced death.
Our results suggested that the anti-inflammatory activity of lycosin-I was associated with NF-κB activation and lycosin-I had potential to be a novel therapeutic candidate for inflammatory diseases.
KeywordsLycosin-I Inflammation HUVECs NF-κB
This project was supported by the Natural Science Foundation of Hunan Province (2015JJ2192 and 2017JJ2372), China Postdoctoral Science Foundation (2015M580704), Scientific Research Foundation of Central South University (2014JSJJ027), the National Natural Science Foundation of China (30901874 and 31672290) and National Undergraduate Innovation Training Program of Central South University (201510533321 and 201610533535). The authors are grateful to Dr. Songping, Liang and Zhonghua Liu, Hunan Normal University, for providing professional advice.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
Research involving human participants and/or animals
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.
- 3.Libby P. Inflammation in atherosclerosis. arteriosclerosis, thrombosis, and vascular biology. 2012;32(9):2045–51. https://doi.org/10.1161/atvbaha.108.179705.
- 5.Kinlay S, Ganz P. Role of endothelial dysfunction in coronary artery disease and implications for therapy. Am J Cardiol. 1997;80(9A):11I-6I.Google Scholar
- 10.Chang CC, Chu CF, Wang CN, Wu HT, Bi KW, Pang JH, et al. The anti-atherosclerotic effect of tanshinone IIA is associated with the inhibition of TNF-alpha-induced VCAM-1, ICAM-1 and CX3CL1 expression. Phytomed: Int J Phytother Phytopharmacol. 2014;21(3):207 – 16. https://doi.org/10.1016/j.phymed.2013.09.012.CrossRefGoogle Scholar
- 12.de Haij S, Bakker AC, van der Geest RN, Haegeman G, Vanden Berghe W, Aarbiou J, et al. NF-kappaB mediated IL-6 production by renal epithelial cells is regulated by c-jun NH2-terminal kinase. J Am Soc Nephrol: JASN. 2005;16(6):1603–11. https://doi.org/10.1681/asn.2004090781.CrossRefPubMedGoogle Scholar
- 13.Boisvert WA, Santiago R, Curtiss LK, Terkeltaub RA. A leukocyte homologue of the IL-8 receptor CXCR-2 mediates the accumulation of macrophages in atherosclerotic lesions of LDL receptor-deficient mice. J Clin Investig. 1998;101(2):353–63. https://doi.org/10.1172/jci1195.CrossRefPubMedPubMedCentralGoogle Scholar
- 17.Baeuerle PA, Henkel T. Function and activation of NF-kappa B in the immune system. Ann Rev Immunol. 1994;12:141–79. https://doi.org/10.1146/annurev.iy.12.040194.001041.CrossRefGoogle Scholar
- 19.Newton K, Dixit VM. Signaling in innate immunity and inflammation. Cold Spring Harbor Perspect Biol. 2012;4(3). https://doi.org/10.1101/cshperspect.a006049.
- 23.Abubakar II, Tillmann T, Banerjee A. Global, regional, and national age-sex specific all-cause and cause-specific mortality for 240 causes of death, 1990–2013: a systematic analysis for the Global Burden of Disease Study 2013. Lancet. 2015;385(9963):117–71. https://doi.org/10.1016/s0140-6736(14)61682-2.CrossRefGoogle Scholar
- 27.Wang L, Wang YJ, Liu YY, Li H, Guo LX, Liu ZH, et al. In vitro potential of Lycosin-I as an alternative antimicrobial drug for treatment of multidrug-resistant Acinetobacter baumannii infections. Antimicrobial Agents Chemother. 2014;58(11):6999–7002. https://doi.org/10.1128/AAC.03279-14.CrossRefGoogle Scholar
- 28.Huang L, Tang Y, Qin J, Peng Y, Yuan Q, Zhang F, et al. Vasoactive intestinal peptide enhances TNF-alpha-induced IL-6 and IL-8 synthesis in human proximal renal tubular epithelial cells by NF-kappaB-dependent mechanism. Inflammation. 2012;35(3):1154–60. https://doi.org/10.1007/s10753-011-9423-4.CrossRefPubMedGoogle Scholar
- 37.Mako V, Czucz J, Weiszhar Z, Herczenik E, Matko J, Prohaszka Z, et al. Proinflammatory activation pattern of human umbilical vein endothelial cells induced by IL-1beta, TNF-alpha, and LPS. Cytometry Part A J Int Soc Anal Cytol. 2010;77(10):962–70. https://doi.org/10.1002/cyto.a.20952.CrossRefGoogle Scholar
- 46.Wang L, Xu Y, Yu Q, Sun Q, Gu Q, Xu X. H-RN, a novel antiangiogenic peptide derived from hepatocyte growth factor inhibits inflammation in vitro and in vivo through PI3K/AKT/IKK/NF-kappaB signal pathway. Biochem Pharmacol. 2014;89(2):255–65. https://doi.org/10.1016/j.bcp.2014.02.026.CrossRefPubMedGoogle Scholar
- 47.Wan M, Liu J, Ouyang X. Nucleotide-binding oligomerization domain 1 regulates Porphyromonas gingivalis-induced vascular cell adhesion molecule 1 and intercellular adhesion molecule 1 expression in endothelial cells through NF-kappaB pathway. J Periodontal Res. 2015;50(2):189–96. https://doi.org/10.1111/jre.12192.CrossRefPubMedGoogle Scholar
- 50.Kurokouchi K, Kambe F, Kikumori T, Sakai T, Sarkar D, Ishiguro N, et al. Effects of glucocorticoids on tumor necrosis factor alpha-dependent activation of nuclear factor kappaB and expression of the intercellular adhesion molecule 1 gene in osteoblast-like ROS17/2.8 cells. J Bone Miner Res. 2000;15(9):1707–15. https://doi.org/10.1359/jbmr.2000.15.9.1707.CrossRefPubMedGoogle Scholar
- 51.Eberhardt W, Schulze M, Engels C, Klasmeier E, Pfeilschifter J. Glucocorticoid-mediated suppression of cytokine-induced matrix metalloproteinase-9 expression in rat mesangial cells: involvement of nuclear factor-kappaB and Ets transcription factors. Mol Endocrinol (Baltimore. Md). 2002;16(8):1752–66. https://doi.org/10.1210/me.2001-0278.CrossRefGoogle Scholar
- 52.Vital AL, Goncalo M, Cruz MT, Figueiredo A, Duarte CB, Lopes MC. Dexamethasone prevents granulocyte-macrophage colony-stimulating factor-induced nuclear factor-kappaB activation, inducible nitric oxide synthase expression and nitric oxide production in a skin dendritic cell line. Mediators Inflammation. 2003;12(2):71–8. https://doi.org/10.1080/0962935031000097673.CrossRefGoogle Scholar
- 53.Ma W, Gee K, Lim W, Chambers K, Angel JB, Kozlowski M, et al. Dexamethasone inhibits IL-12p40 production in lipopolysaccharide-stimulated human monocytic cells by down-regulating the activity of c-Jun N-terminal kinase, the activation protein-1, and NF-kappa B transcription factors. J Immunol (Baltimore: 1950). 2004;172(1):318–30.Google Scholar
- 54.Mattioli I, Sebald A, Bucher C, Charles RP, Nakano H, Doi T, et al. Transient and selective NF-kappa B p65 serine 536 phosphorylation induced by T cell costimulation is mediated by I kappa B kinase beta and controls the kinetics of p65 nuclear import. J Immunol (Baltimore: 1950). 2004;172(10):6336–44.Google Scholar