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Current Drug Discovery Technologies

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ISSN (Print): 1570-1638
ISSN (Online): 1875-6220

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Research Article

The Immunomodulatory Role of G2013 (α-L-Guluronic Acid) on the Expression of TLR2 and TLR4 in HT29 cell line

Author(s): Hamid Farhang, Laleh Sharifi, Mohammad Mehdi Soltan Dallal, Mona Moshiri, Zahra Norouzbabaie, Saied Bokaie, Somaye Aletaha, Seyed Jalal Zargar and Abbas Mirshafiey*

Volume 16, Issue 1, 2019

Page: [91 - 95] Pages: 5

DOI: 10.2174/1570163815666180226093711

Price: $65

Abstract

Background: The non-steroidal anti-inflammatory drugs (NSAIDs) play crucial role in the controlling of inflammatory diseases. Due to the vast side effects of NSAIDs, its use is limited. G2013 or α-L-Guluronic Acid is a new NSAID with immunomodulatory features. Objectives: Considering the leading role of TLRs in inflammatory responses, in this study, we aimed to evaluate G2013 cytotoxicity and its effect on the expression of TLR2 and TLR4 molecules.

Methods: HEK293-TLR2 and HEK293-TLR4 cells were cultured and seeded on 96-well cell plate, and MTT assay was performed for detecting the viability of the cells after treatment with different concentrations of G2013. HT29 cells were grown and treated with low and high doses of G2013. After total RNA extraction and cDNA synthesis, quantitative real-time PCR were performed to assess the TLR2 and TLR4 mRNA synthesis.

Results: We found that concentrations of ≤125 µg/ml of G2013 had no apparent cytotoxicity effect on the HEK293-TLR2 and -TLR4 cells. Our results indicated that after G2013 treatment (5 µg/ml) in HT29 cells, TLR2 and TLR4 mRNA expression decreased significantly compared with the untreated control group (p=0.02 and p=0.001 respectively).

Conclusion: The results of this study revealed that G2013 can down regulate the TLR2 and TLR4 gene expression and exerts its inhibitory effect. Our findings are parallel to our previous finding which showed G2013 ability to down regulate the signaling pathway of TLRs. However, further studies are needed to identify the molecular mechanism of G2013.

Keywords: G2013, α-L-Guluronic Acid, NSAIDs, TLR2, TLR4, HT29.

Graphical Abstract

[1]
Zhao S, Zhang Y, Zhang Q, Wang F, Zhang D. Toll-like receptors and prostate cancer. Front Immunol 2014; 5: 352.
[2]
Takeda K, Kaisho T, Akira S. Toll-like receptors. Annu Rev Immunol 2003; 21(1): 335-76.
[3]
Janeway CA. Cold Spring Harbor symposia on quantitative biology. Cold Spring Harbor Laboratory Press 1989; pp. 1-13.
[4]
Huang L-Y, DuMontelle JL, Zolodz M, Deora A, Mozier NM, Golding B. Use of toll-like receptor assays to detect and identify microbial contaminants in biological products. J Clin Microbiol 2009; 47(11): 3427-34.
[5]
Lee MS, Kim Y-J. Signaling pathways downstream of pattern-recognition receptors and their cross talk. Annu Rev Biochem 2007; 76: 447-80.
[6]
Joosten LA, Abdollahi-Roodsaz S, Dinarello CA, O’neill L, Netea MG. Toll-like receptors and chronic inflammation in rheumatic diseases: new developments. Nat Rev Rheumatol 2016; 12(6): 344-57.
[7]
De Jager PL, Franchimont D, Waliszewska A, et al. The role of the Toll receptor pathway in susceptibility to inflammatory bowel diseases. Genes Immun 2007; 8(5): 387-97.
[8]
Liu Y, Yin H, Zhao M, Lu Q. TLR2 and TLR4 in autoimmune diseases: a comprehensive review. Clin Rev Allergy Immunol 2014; 47(2): 136-47.
[9]
Sidiropoulos P, Hatemi G, Song I-H, et al. Evidence-based recommendations for the management of ankylosing spondylitis: systematic literature search of the 3E Initiative in Rheumatology involving a broad panel of experts and practising rheumatologists. Rheumatology 2008; 47(3): 355-61.
[10]
Poddubnyy D. Axial spondyloarthritis: is there a treatment of choice? Ther Adv Musculoskelet Dis 2013; 5(1): 45-54.
[11]
Crofford LJ. Use of NSAIDs in treating patients with arthritis. Arthritis Res Ther 2013; 15(3): S2.
[12]
Vitiello M, Abuchar A, Santana N, Dehesa L, Kerdel FA. An update on the treatment of the cutaneous manifestations of systemic sclerosis: The dermatologist’s point of view. J Clin Aesthet Dermatol 2012; 5(7): 33-43.
[13]
Østensen M, Villiger P. Nonsteroidal anti-inflammatory drugs in systemic lupus erythematosus. Lupus 2001; 10(3): 135-9.
[14]
Watanabe T, Higuchi K, Kobata A, et al. Non-steroidal anti-inflammatory drug-induced small intestinal damage is Toll-like receptor 4 dependent. Gut 2008; 57(2): 181-7.
[15]
Page TH, Turner JJ, Brown AC, et al. Nonsteroidal anti-inflammatory drugs increase TNF production in rheumatoid synovial membrane cultures and whole blood. J Immunol 2010; 185(6): 3694-701.
[16]
Fattahi MJ, Abdollahi M, Agha Mohammadi A, et al. Preclinical assessment of β-d-mannuronic acid (M2000) as a non-steroidal anti-inflammatory drug. Immunopharmacol Immunotoxicol 2015; 37(6): 535-40.
[17]
Mirshafiey A, Hosseini S, Afraei S, Rastkari N. T Zavareh F, Azizi G. Anti-aging property of G2013 molecule as a novel immunosuppressive agent on enzymatic and non-enzymatic oxidative stress determinants in rat model. Curr Drug Discov Technol 2016; 13(1): 25-33.
[18]
Nazeri S, Khadem Azarian S, Fattahi MJ, et al. Preclinical and pharmacotoxicology evaluation of α-l-guluronic acid (G2013) as a non-steroidal anti-inflammatory drug with immunomodulatory property. Immunopharmacol Immunotoxicol 2017; 39(2): 59-65.
[19]
Fattahi MJ, Abdollahi M, Agha Mohammadi A, et al. Preclinical assessment of beta-D-mannuronic acid (M2000) as a non-steroidal anti-inflammatory drug. Immunopharmacol Immunotoxicol 2015; 37(6): 535-40.
[20]
Afraei S, Azizi G, Zargar SJ, Sedaghat R, Mirshafiey A. New therapeutic approach by G2013 in experimental model of multiple sclerosis. Acta Neurol Belg 2015; 115(3): 259-66.
[21]
Mirshafiey A, Hosseini S, Afraei S, Rastkari N, Zavareh FT, Azizi G. Anti-aging property of G2013 molecule as a novel immunosuppressive agent on enzymatic and non-enzymatic oxidative stress determinants in Rat model. Curr Drug Discov Technol 2016; 13(1): 25-33.
[22]
Nazeri S, Khadem Azarian S, Fattahi MJ, et al. Preclinical and pharmacotoxicology evaluation of alpha-l-guluronic acid (G2013) as a non-steroidal anti-inflammatory drug with immunomodulatory property. Immunopharmacol Immunotoxicol 2017; 39(2): 59-65.
[23]
Arjomand Fard N, Tabrizian N, Mirzaei R, et al. Efficacy and safety of G2013 as a novel immunosuppressive agent on differentiation, maturation and function of human dendritic cells. Iran J Public Health 2017; 46(2): 216-21.
[24]
Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)). Method Methods 2001; 25(4): 402-8.
[25]
Sidiropoulos PI, Hatemi G, Song IH, et al. Evidence-based recommendations for the management of ankylosing spondylitis: Systematic literature search of the 3E Initiative in Rheumatology involving a broad panel of experts and practising rheumatologists. Rheumatology 2008; 47(3): 355-61.
[26]
Afraei S, Azizi G, Zargar SJ, Sedaghat R, Mirshafiey A. New therapeutic approach by G2013 in experimental model of multiple sclerosis. Acta Neurol Belg 2015; 115(3): 259-66.
[27]
Hajivalili M, Pourgholi F, Majidi J, et al. G2013 modulates TLR4 signaling pathway in IRAK-1 and TARF-6 dependent and miR-146a independent manner. Mol Cell Biol 2016; 62(4): 1-5.
[28]
Mortazavi JSS, Yousefi M, Sadat NS, Motamed N, Tofighi ZF, Mirshafiey A. Inhibitory effect of G2013 molecule as a novel immunomodulatory agent, on miR-155 gene expression in HEK-Blue hTLR4 cell line. Eur J Inflamm 2016; 14(2): 86-92.
[29]
Sharifi L, Mohsenzadegan M, Aghamohammadi A, et al. Immunomodulatory effect of G2013 (a-L-Guluronic acid) on theTLR2 and TLR4 in human mononuclear cells. Curr Drug Discov Technol 2018. [Epub ahead of print].
[30]
Sharifi L, Mohsenzadegan M, Rezaei N, et al. Immunomodulation of TLR2 and TLR4 by G2013 (alpha-L-guluronic acid) in CVID patients. Int J Pediatr 2017; 5(7): 5327-37.

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