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http://dx.doi.org/10.7732/kjpr.2020.33.3.183

Immune-Enhancing Effects of Green Lettuce (Lactuca sativa L.) Extracts through the TLR4-MAPK/NF-κB Signaling Pathways in RAW264.7 Macrophage Cells  

Seo, Hyun-Ju (Institute of Agricultural Science and Technology, Andong National University)
Jeong, Jin Boo (Department of Medicinal Plant Resources, Andong National University)
Publication Information
Korean Journal of Plant Resources / v.33, no.3, 2020 , pp. 183-193 More about this Journal
Abstract
Recently, as a natural substance has been emphasized interest in research to enhance the immune function. Green lettuce (Lactuca sativa L.) is a popular vegetable used fresh and it contains various phytochemicals and antioxidant compounds, and has been reported to have various physiological activities such as antibacterial, antioxidant, antitumor and anti-mutagenic. However, only a few studies have investigated on the mechanism of action of immune-enhancing activity of lettuce. Therefore, in this study, the immunomodulatory activities and potential mechanism of action of Green lettuce extracts (GLE) were evaluated in the murine macrophage cell line RAW264.7. GLE significantly increased NO levels by RAW264.7 cells, as well as expressions of immunomodulators such as iNOS, COX-2, IL-1β, IL-6, IL-12, TNF-α and MCP-1. Although GLE activated ERK1/2, p38, JNK and NF-κB, GLE-mediated expressions of immunomodulators was dependent on p38, JNK and NF-κB. In addition, TLR4 inhibition blocked GLE-mediated expressions of immunomodulators and activation of p38, JNK and NF-κB. Taken together, these results demonstrated that TLR4-MAPK/NF-κB signalling pathways participated in GLE-induced macrophage activation and GLE could be developed as a potential immunomodulating functional food.
Keywords
Green lettuce; Immune-enhancing effects; Lactuca sativa L.; Macrophage activation;
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1 Xi, L., C. Xiao and R.H.J. Bandsma. 2011. C-reactive protein impairs hepatic insulin sensitivity and insulin signaling in rats: role of mitogen-activated protein kinases. Hepatology 53:127-135.   DOI
2 Yu, Y., M.Y. Shen, Z.J. Wang, Y.X. Wang, M.Y. Xie and J.H. Xie. 2017. Sulfated polysaccharide from Cyclocarya paliurus enhances the immunomodulatory activity of macrophages. Carbohydr. Polym. 174:669-676.   DOI
3 Zhang, M., W. Wu, Y. Ren, X. Li, Y. Tang, T. Min, F. Lai and H.Wu. 2017. Structural characterization of a novel polysaccharide from Lepidium meyenii (Maca) and analysis of its regulatory function in macrophage polarization in vitro. J. Agric. Food Chem. 65:1146-1157.   DOI
4 Zheng, D.H., Y. Zhou, S.J. Cobbina, W. Wang, Q. Li and Y. Chen. 2017. Purification, characterization, and immunoregulatory activity of a polysaccharide isolated from Hibiscus sabdariffa L. J. Sci. Food Agric. 97(5):1599-1606.   DOI
5 Lechner, M., P. Lirk and J. Rieder. 2005. Inducible nitric oxide synthase (iNOS) in tumor biology: the two sides of the same coin. Semin. Cancer Biol. 15:277-289.   DOI
6 Pautz, A., J. Art, S. Hahn, S. Nowag, C. Voss and H. Kleinert. 2010. Regulation of the expression of inducible nitric oxide synthase. Nitric Oxide 23:75-93.   DOI
7 Li, F., X.Y. Sun, X.W. Li, T. Yang and L.W. Qi. 2017. Enrichment and separation of quercetin-3-O-d-glucuronide from lotus leaves (Nelumbo nucifera gaertn.) and evaluation of its anti-inflammatory effect. J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci. 1040:186-191.   DOI
8 Liu, X., J.H. Xie, S. Jia, L.X. Huang, Z.J. Wang and C. Li. 2017. Immunomodulatory effects of an acetylated Cyclocarya paliurus polysaccharide on murine macrophages RAW264.7. Int. J. Biol. Macromol. 98:576-581.   DOI
9 Lo, A.H., Y.C. Liang, S.Y. Lin-Shiau, C.T. Ho and J.K. Lin. 2002. Carnosol, an antioxidant in rosemary, suppresses inducible nitric oxide synthase through down-regulating nuclear factor-kappaB in mouse macrophages. Carcinogenesis 23(6):983-991.   DOI
10 Ma, W. K. Gee, W. Lim, K. Chambers, J.B. Angel and M. Kozlowski. 2004. 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. 172:318-330.   DOI
11 Materska, M., K. Olszowka, B. Chilczuk, A. Stochmal, L. Pecio, B. Pacholczyk-Sienicka, S. Piacente, C. Pizza and M. Masullo. 2019. Polyphenolic profiles in lettuce (Lactuca sativa L.) after $CaCl_2$ treatment and cold storage. Eur. Food Res. Technol. 245:733-744.   DOI
12 Mitchell, J.A., S. Larkin and T.J. Williams. 1995. Cyclooxygenase-2: Regulation and relevance in inflammation. Biochem. Pharmacol. 50:1535-1542.   DOI
13 Mitchell, R.A., H. Liao, J. Chesney, G. Fingerle-Rowson, J. Baugh, I. David and R. Bucala. 2002. Macrophage migration inhibitory factor (MIF) sustains macrophage proinflammatory function by inhibiting p53: Regulatory role in the innate immune response. PNASU. 99(1):345-350.   DOI
14 Natarajan, K., P. Abraham, R. Kota and B. Isaac. 2018. NF-${\kappa}$ B-iNOS-COX2-TNF ${\alpha}$ inflammatory signaling pathway plays an important role in methotrexate induced small intestinal injury in rats. Food Chem. Toxicol. 118:766-783.   DOI
15 Adesso, S., G. Pepe, E. Sommella, M. Manfra, A. Scopa, A. Sofo, G.C. Tenore, M. Russo, F. Di Gaudio, G. Autore, P. Campiglia and S. Marzocco. 2016. Anti-inflammatory and antioxidant activity of polyphenolic extracts from Lactuca sativa (var. Maravilla de Verano) under different farming methods. J. Sci. Food Agric. 96:4194-4206.   DOI
16 Akira, S. 2009. Innate immunity to pathogens: diversity in receptors for microbial recognition. Immunol. Rev. 227:5-8.   DOI
17 Arancibia, S.A., C.J. Beltran, I.M. Aguirre, P. Silva, A.L. Peralta, F. Malinarich and M.A. Hermoso. 2007. Toll-like receptors are key participants in innate immune responses. Biol. Res. 40:97-112.   DOI
18 Belardelli, F. 1995. Role of interferons and other cytokines in the regulation of the immune response. APMIS. 103(1-6):161-179.   DOI
19 Bosca, L., M. Zeini, P.G. Traves and S. Hortelano. 2005. Nitric oxide and cell viability in inflammatory cells: A role for NO in macrophage function and fate. Toxicol 208(2):249-258.   DOI
20 Brubaker, S.W., K.S. Bonham, I. Zanoni and J.C. Kagan. 2015. Innate immune pattern recognition: A cell biological perspective. Annu. Rev. Immunol. 33:257-290.   DOI
21 Chardin, C., S.T. Schenk, H. Hirt, J. Colcombet and A. Krapp. 2017. Review: Mitogen-activated protein kinases in nutritionalsignaling inarabidopsis. Plant Sci. 260:101-108.   DOI
22 Chehimi, J., S.E. Starr, I. Frank, A. D'Andrea, X. Ma, R.R. MacGregor,J. Sennelier and G. Trinchieri. 1994. Impaired interleukin 12 production in human immunodeficiency virus-infected patients. J. Exp. Med. 179:1361-1366.   DOI
23 Chen, F., V. Castranova and X. Shi. 2001. New insights into the role of nuclear factor-${\kappa}$B in cell growth regulation. Am. J. Pathol. 159(2):387-397.   DOI
24 Newstead, S.L., A.J. Gates, M.G. Hartley, C.A. Rowland, E.D. Williamson and R.A. Lukaszewski. 2014. Control of intracellular Francisella tularensis by different cell types and the role of nitric oxide. J. Immunol. Res. 694-717.
25 Nie, C., P. Zhu, S. Ma, M. Wang and Y. Hu. 2018. Purification, characterization and immunomodulatory activity of polysaccharides from stem lettuce. Carbohydr. Polym. 188:236-242.   DOI
26 Oh, H.M., E.E. Carey and C.B. Rajashekar. 2009. Environmental stresses induce health-promoting phytochemicals in lettuce. Plant Physiol. Biochem. 47:578-583.   DOI
27 Ohta, Y., J.B. Lee, K. Hayashi, A. Fujita, D.K. Park and T. Hayashi. 2007. In vivo anti-influenza virus activity of an Immunomodulatory Acidic Polysaccharide Isolated from Cordyceps militarisGrown on Germinated Soybeans. J. Agric. Food Chem. 55(25):10194-10199.   DOI
28 O'Neill, L.A.J., D. Golenbock and A.G. Bowie. 2013. The history of Toll-like receptors redefining innate immunity. Nat. Rev. Immunol. 13:453-460.   DOI
29 Palmer, R.M.J., D.S. Ashton and S. Moncada, 1988. Vascular endothelial cells synthesize nitric oxide from L-arginine. Nature 333(6174):664-666.   DOI
30 Park, H.J. 2019. Isolation of the constituent inhibiting nitric oxide formation from Lycopus lucidus in LPS-induced macrophage cells. Korean J. Plant Res. 32(4):264-269.
31 Pearson, G., F. Robinson, T. Beers Gibson, B.E. Xu, M. Karandikar, K. Berman and M.H. Cobb. 2001. Mitogenactivated protein (MAP) kinase pathways: Regulation and physiological functions. Endocr. Rev. 22(2):153-183.   DOI
32 Pluddemann, A., S. Mukhopadhyay and S. Gordon. 2011. Innate immunity to intracellular pathogens: macrophage receptors and responses to microbial entry. Immunol. Rev. 240:11-24.   DOI
33 Chen, W.X., W.Y. Zhang, W.B. Shen and K.C. Wang. 2010. Effects of the acid polysaccharide fraction isolated from a cultivated Cordyceps sinensison macrophages in vitro. Cell Immunol. 262(1):69-74.   DOI
34 Cho, J.W., K.S. Lee and C.W. Kim. 2007. Curcumin attenuates the expression of IL-1${\beta}$, IL-6, and TNF-${\alpha}$ as well as cyclin E in TNF-${\alpha}$-treated HaCaT cells; NF-${\kappa}$B and MAPKs as potential upstream targets. Int. J. Mol. Med. 19:469-474.
35 Cho, Y.C., T.T. Bach, B.R. Kim, H.L. Vuong and S. Cho. 2017. Spilanthes acmella inhibits inflammatory responses via inhibition of NF-${\kappa}$B and MAPK signaling pathways in RAW 264.7 macrophages. Mol. Med. Rep. 16:339-346.   DOI
36 Crozier, A, M.E.J. Lean, M.S. McDonald and C. Black. 1997. Quantitative analysis of the flavonoid content of commercial tomatoes, onions, lettuce and celery. J. Agric. Food Chem. 45:590-595.   DOI
37 Degl'Innoocenti, E., A. Pardossi, M. Tattini and L. Guidi. 2008. Phenolic compounds and antioxidant power in minimally processed salad. J. Food Biochem. 32:642-653.   DOI
38 Delgado, A .V., A .T. McManus a nd J .P. Chambers. 20 0 3. Production of tumor necrosis factor-alpha, interleukin 1-beta, interleukin 2, and interleukin 6 by rat leukocyte subpopulations after exposure to substance P. Neuropeptides 37(6):355-361.   DOI
39 Deretic, V., T. Saitoh and S. Akira. 2013. Autophagy in infection, inflammation and immunity. Nat. Rev. Immunol. 13:722-737.   DOI
40 Drummond, R.A. and G.D. Brown. 2011. The role of Dectin-1 in the host defence against fungal infections. Curr. Opin. Microbiol. 14:392-399.   DOI
41 Ren, H, J. Hao, T. Liu, D. Zhang, H. Lv, E. Song and C. Zhu. 2016. Hesperetin suppresses inflammatory responses in lipopolysaccharide-induced RAW 264.7 cells via the inhibition of NF-${\kappa}$B and activation of Nrf2/HO-1 pathways. Inflammation 39:964-973.
42 Romani, A., P. Pinelli, C. Galardi, G. Sani, A. Cimato and D. Heimler. 2002. Polyphenols in greenhouse and open-airgrown lettuce. Food Chem. 79:337-342.   DOI
43 Sfondrini, L, Balsari, A and Menard, S. 2003. Innate immunity in breast carcinoma. Endocr. Relat. Cancer 10:301-308.   DOI
44 Sherwin, C.and R. Fern. 2005. Acute lipopolysaccharidemediated injury in neonatal white matter glia: Role of TNFalpha, IL-1beta, and calcium. J. Immunol. 175(1):155-161.   DOI
45 Shi, C., and E.G. Pamer. 2011. Monocyte recruitment during infection and inflammation. Nat. Rev. Immunol. 11(11):762-774.   DOI
46 Singh, R.K.and I.J. Fidler. 1993. Monocyte chemotactic and activating factor and lipopolysaccharide for activation of antitumor properties in human blood monocytes. Lymphokine Cytokine Res. 2:285-291.
47 Picot, D., P.J. Loll and R.M. Garavito. 1994. The X-ray crystal structure of the membrane protein prostaglandin H2 synthase-1. Nature 367:243-249.   DOI
48 Smith, W.L., R.M. Garavito and D.L. DeWitt. 1996. Prostaglandin Endoperoxide H Synthases (Cyclooxygenases)-1 and -2. J. Biol. Chem. 271:33157-33160.   DOI
49 Suh, S.J., U.H. Jin, K.W. Kim, H.W. Chang, Y.C. Lee and C.H. Kim. 2007. Triterpenoid saponin, oleanolic acid 3-Obeta-dglucopyranosyl(1$\rightarrow$3)-alpha-l-rhamnopyranosyl(1$\rightarrow$2)-alpha-l- arabinopyranoside (OA) from Aralia elata inhibits LPS-induced nitric oxide production by down-regulated NF-kappaB in raw 264.7 cells. Arch. Biochem. Biophys. 467(2):227-33.   DOI
50 DuPont, M.S., Z. Mondin, G. Williamson and K.R. Price. 2000. Effect of variety, processing, and storage on the flavonoid glycoside content and composition of lettuce and endive. J. Agric. Food Chem. 48:3957-3964.   DOI
51 El-Seedi, H.R., A.M.A. El-Said, S.A.M. Khalifa, U. Goransson, L. Bohlin, A.K. Borg-Karlson and R. Verpoorte. 2012. Biosynthesis, natural sources, dietary intake, pharmacokinetic properties, and biological activities of hydroxycinnamic acids. J. Agric. Food Chem. 60:10877-10895.   DOI
52 Falch, B.H., T. Espevik, L. Ryan and B.T. Stokke. 2000. The cytokine stimulating activity of (1$\rightarrow$3)-${\beta}$-D-glucans is dependent on the triple helix conformation. Carbohydr. Res. 329:587-596.   DOI
53 Fisher, W.G., P.C. Yang, R.K. Medikonduri and M. Saleet Jafri. 2006. NFAT and NF${\kappa}$B activation in T lymphocytes: A model of differential activation of gene expression. Ann. Biomed. Eng. 34:1712-1728.   DOI
54 Garlanda, C, C.A. Dinarello and A. Mantovani. 2013. The Interleukin-1 Family: Back to the Future. Immunity 39:1003-1008.   DOI
55 Giampieri, F., J.M. Alvarez-Suarez and M. Battino. 2014. Strawberry and human health: effects beyond antioxidant activity. J. Agric. Food Chem. 62:3867-3876.   DOI
56 Gordon, B.H., M.S. Kang, P. Cho and K.P. Sucher. 2000. Dietary habits and health beliefs of Korean-Americans in the San Francisco Bay Area. J. Am. Diet Assoc. 100:1198-1201.   DOI
57 Gu, L., S.C. Tseng and B.J. Rollins. 1999. Monocyte chemoattractant protein-1. Chem. Immunol. 72:7-29.   DOI
58 Strieter, R.M., S.L. Kunkel and R.C. Bone. 1993. Role of tumor necrosis factor-alpha in disease states and inflammation. Crit. Care Med. 21(10):S447-463.   DOI
59 Terazawa, R., N. Akimoto, T. Kato, T. Itoh, Y. Fujita, N. Hamada, T. Deguchi,M. Iinuma,M. Noda, Y. Nozawa and M. Ito. 2013. A kavalactone derivative inhibits lipopolysaccharide- stimulated iNOS induction and NO production through activation of Nrf2 signaling in BV2 microglial cells. Pharmacol. Res. 71:34-43.   DOI
60 Tilman, D. and M. Clark. 2014. Global diets link environmental sustainability and human health. Nature 515:518-522.   DOI
61 Trinchieri, G. 2003 Interleukin-12 and the regulation of innate resistance and adaptive immunity. Nat. Rev. Immunol. 3:133-146.   DOI
62 Vane, J.R., J.A. Mitchell, I. Appleton, A. Tomlinson, D. Bishop-Bailey, J. Croxtall and D.A. Willoughby. 1994. Inducible isoforms of cyclooxygenase and nitric-oxide synthase in inflammation. Proc. Natl. Acad. Sci. USA 91(6):2046-2050.   DOI
63 Vitenberga, Z. and M. Pilmane. 2017. Inflammatory, antiinflammatory and regulatory cytokines in relatively healthy lung tissue as an essential part of the local immune system. Biomed. Pap. Med. Fac. Univ. Palacky. Olomouc. Czech. Repub. 161(2):164-173.   DOI
64 Wen, Z.S., X.W. Xiang, H.X. Jin, X.Y. Guo, L.J. Liu, L.J. and Y.N. Huang. 2016. Composition and anti-inflammatory eff ect of polysaccharides from Sargassum horneri in Raw264.7 macrophages. Int. J. Biol. Macromol. 88:403-413.   DOI
65 Wieczynska, J. and I. Cavoski. 2018. Antimicrobial, antioxidant and sensory features of eugenol, carvacrol and trans-anethole in active packaging for organic ready-to-eat iceberg lettuce. Food Chem. 259:251-260.   DOI
66 Guo, F., H. He, Z.C. Fu, S. Huang, T. Chen, C.J. Papasian, L.R. Morse, Y. Xu, R.A. Battaglino, X.F. Yang, Z. Jiang, H.B. Xin and M. Fu. 2015. Adipocyte-derived PAMM suppresses macrophage inflammation by inhibiting MAPK signalling. Biochem. J. 472:309-318.   DOI
67 Hawkey, C.J. 1999. COX-2 inhibitors. Lancet 353:307-314.   DOI
68 Hayden, M.S. and S. Ghosh. 2008. Shared principles in NF-${\kappa}$B signaling. Cell 132:344-362.   DOI
69 Hedges, L.J. and C.E. Lister. 2005. Nutritional attributes of salad vegetables. Crop & Food Research Confidential Report NO. 1473:1-29.
70 Hommes, D.W., M.P. Peppelenbosch and S.J. van Deventer. 2003. Mitogen activated protein (MAP) kinase signal transduction pathways and novel anti-inflammatory targets. Gut 52(1):144-151.   DOI
71 Junt, T., E.A. Moseman, M. Iannacone, S. Massberg, P.A. Lang, M. Boes and U.H. von Andrian. 2007. Subcapsular sinus macrophages in lymph nodes clear lymph-borne viruses and present them to antiviral B cells. Nature 450(7166):110-114.   DOI
72 Kawamoto, T., M. Ii, T. Kitazaki, Y. Iizawa and H. Kimura. 2008. TAK-242 selectively suppresses Toll-like receptor 4-signaling mediated by the intracellular domain. Eur. J. Pharmacol. 584(1):40-48.   DOI
73 Kawai, T. and S. Akira. 2010. The role of pattern-recognition receptors in innate immunity:update on Toll-like receptors. Nat. Immunol. 11:373-384.   DOI
74 Kesteloot, H. and J. Zhang. 2000. Salt consumption during the nutrition transition in South Korea. Am. J. Clin. Nutr. 72:199-201.   DOI
75 Kikuchi, K., Y. Yanagawa, T. Aranami, C. Iwabuchi, K. Iwabuchi and K. Onoe. 2003. Tumour necrosis factor-${\alpha}$ but not lipopolysaccharide enhances preference of murine dendritic cells for Th2 differentiation. Immunology 108(1):42-49.   DOI
76 Kopitar-Jerala, N. 2015. Innate immune response in brain, NF-kappa B signaling and cystatins. Front. Mol. Neurosci. 8:216-220.   DOI
77 Labonte, A.C., A.C. Tosello-Trampont and Y.S. Hahn. 2014. The role of macrophage polarization in infectious and inflammatory diseases. Mol. Cells 37:275-285.   DOI
78 Lamkanfi, M. and V.M. Dixit. 2014. Mechanisms and functions of inflammasomes. Cell 157:1013-1022.   DOI
79 Lankford, C.S.R and D.M. Frucht. 2003. A unique role for IL-23 in promoting cellular immunity. J. Leukoc. Biol. 73:49-56.   DOI
80 Lee, J, K. Demissie, S.E. Lu and G.G. Rhoads. 2007. Cancer incidence among Korean-American immigrants in the United States and native Koreans in South Korea. Cancer Control 14:78-85.   DOI
81 Lee, S.B., W.S. Lee, J.S. Shin, D.S. Jang and K.T. Lee. 2017. Xanthotoxin suppresses LPS-induced expression of iNOS, COX-2, TNF-${\alpha}$, and IL-6 via AP-1, NF-${\kappa}$B, and JAK-STAT inactivation in RAW 264.7 macrophages. Int. Immunopharmacol. 49:21-29.   DOI
82 Lei, W., J.D. Browning, P.A. Eichen, C.H. Lu, V.V. Mossine, G.E. Rottinghaus, W.R. Folk, G.Y. Sun, D.B. Lubahn and K.L. 2015. Fritsche, immuno-stimulatory activity of a polysaccharide-enriched fraction of Sutherlandia frutescens occurs by the toll-like receptor-4 signalling pathway. J. Ethnopharmacol. 172:247-253.   DOI