Thiazolidinedione Derivative Suppresses LPS-induced COX-2 Expression and NO Production in RAW 264.7 Macrophages

Document Type : Research article

Authors

1 Department of Biology, Faculty of Science, University of Guilan, University Campus 2, Rasht, Iran.

2 Department of Biology, Faculty of Science, University of Guilan, Rasht, Iran.

Abstract

The present study was designed to investigate the inhibitory effect of 2,4 bis-[(4-ethoxyphenyl)azo] 5-(3-hydroxybenzylidene) thiazolidine-2,4-dione (TZD-OCH2CH3) on the cyclo-oxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in RAW 264.7 cells. The effects of TZD-OCH2CH3 on COX-2 and iNOS mRNA expression in LPS-activated RAW 264.7 cells were detected by real time PCR. Also, to understand structure and substrate specificity, we have utilized molecular docking simulations (AutoDock Vina) and the active residues in the binding pocket were determined from COX-2 and iNOS. The treatment of RAW 264.7 cells with TZD-OCH2CH3 significantly inhibited LPS-induced COX-2 mRNA expression, corresponding to 46.1% and 61.06% at 30 and 60 μg/mL, respectively. The present study revealed that the TZD-OCH2CH3 had a little effect on iNOS mRNA expression. Meanwhile, the TZD-OCH2CH3 also could inhibit the production of NO compared to single LPS-stimulated cell. According to the results obtained, TZD-OCH2CH3 dramatically suppressed lipopolysaccharide (LPS) induced nitric oxide (NO) production after 24 h, in a concentration-dependent manner with an IC50 of 65 μg/mL. Our data suggest that TZD-OCH2CH3, as a functionally novel agent, inhibits the inflammatory pathway via suppression of COX-2 mRNA expression and also by the inhibition of the iNOS activity. Therefore, this compound could be suggested as a novel therapeutic strategy for inflammation-associated disorders.

Keywords

Main Subjects


References
Botting RM. Inhibitors of cyclooxygenases:
mechanisms, selectivity and uses. J. Physiol.
Pharmacol. (2006) 57: 113.
Park JY, Pillinger MH and Abramson SB. Prostaglandin
E 2 synthesis and secretion: the role of PGE 2
synthases. Clin. Immunol. (2006) 119: 229-40.
Kean WF, Rainsford KD and Kean IRL. Management
of chronic musculoskeletal pain in the elderly: opinions
on oral medication use. Inflammopharmacology
(2008) 16: 53-75.
Luo Y, Ma L, Zheng H, Chen L, Li R, He C, Yang L and
Wei Y. Discovery of (Z)-5-(4-methoxybenzylidene)
thiazolidine-2, 4-dione, a readily available and orally
active glitazone for the treatment of concanavalin
A-induced acute liver injury of BALB/c mice. J. Med.
Chem. (2009) 53: 273-81.
Schmidt M, Christiansen CF, Mehnert F, Rothman KJ
and Sørensen HT. Non-steroidal anti-inflammatory
drug use and risk of atrial fibrillation or flutter:
population based case-control study. BMJ (2011) 343:
d3450.
Brown JR and DuBois RN. COX-2: a molecular target
for colorectal cancer prevention. J. Clin. Oncol. (2005)
23: 2840-55.
Coussens LM and Werb Z. Inflammation and cancer.
Nature (2002) 420: 860-70.
Dannenberg AJ, Altorki NK, Boyle JO, Dang
C, Howe LR, Weksler BB and Subbaramaiah K.
Cyclo-oxygenase 2: a pharmacological target for the
prevention of cancer. Lancet Oncol. (2001) 2: 544-51.
Harris RE, Alshafie GA, Abou-Issa H and Seibert K.
Chemoprevention of breast cancer in rats by celecoxib,
a cyclooxygenase 2 inhibitor. Cancer Res. (2000) 60:
2101-3.
Waskewich C, Blumenthal RD, Li H, Stein R,
Goldenberg DM and Burton J. Celecoxib exhibits
the greatest potency amongst cyclooxygenase (COX)
inhibitors for growth inhibition of COX-2-negative
hematopoietic and epithelial cell lines. Cancer Res.
(2002) 62: 2029-33.
Bhatt DL, Scheiman J, Abraham NS, Antman EM,
Chan FK, Furberg CD, Johnson DA, Mahaffey KW
and Quigley EM; American College of Cardiology
Foundation Task Force on Clinical Expert Consensus
Documents. ACCF/ACG/AHA 2008 expert consensus
document on reducing the gastrointestinal risks of
antiplatelet therapy and NSAID use: a report of the
American college of cardiology foundation task force
on clinical expert consensus documents. J. Am. Coll.
Cardiol. (2008) 52: 1502-17.
Lanas A, García-Rodríguez LA, Arroyo MT, Gomollón
F, Feu F, González-Pérez A, Zapata E, Bástida G,
Rodrigo L, Santolaria S, Güell M, de Argila CM,
Quintero E, Borda F, Piqué JM; Asociación Española de
Gastroenterología. Risk of upper gastrointestinal ulcer
bleeding associated with selective cyclo-oxygenase-2
inhibitors, traditional non-aspirin non-steroidal anti-
(1)
(2)
(3)
(4)
(5)
(6)
(7)
(8)
(9)
(10)
(11)
(12)
inflammatory drugs, aspirin and combinations. Gut
(2006) 55: 1731-8.
Beraldo H and Gambinob D. The wide pharmacological
versatility of semicarbazones, thiosemicarbazones and
their metal complexes. Mini-Rev. Med. Chem. (2004)
4: 31-9.
Dua R, Shrivastava S, Sonwane SK and Srivastava SK.
Pharmacological significance of synthetic heterocycles
scaffold: a review. Adv. Biol. Res. (2011) 5: 120-44.
Bhatnagar A, Sharma PK and Kumar N. A review
on imidazoles: Their chemistry and pharmacological
potentials. Int. J. PharmTech Res. (2011) 3: 268-82.
Jain VS, Vora DK and Ramaa CS. Thiazolidine-2,
4-diones: progress towards multifarious applications.
Bioorg. Med. Chem. (2013) 21: 1599-620.
Luo Y, Ma L, Zheng H, Chen L, Li R, He C, Yang S,
Ye X, Chen Z, Li Z, Gao Y, Han J, He G, Yang L and
Wei Y. Discovery of (Z)-5-(4-methoxybenzylidene)
thiazolidine-2, 4-dione, a readily available and orally
active glitazone for the treatment of concanavalin
A-induced acute liver injury of BALB/c mice. J. Med.
Chem. (2009) 53: 273-81.
Minkin VI, Garnovskii AD, Elguero J, Katritzky AR
and Denisko OV. Tautomerism of heterocycles: fivemembered rings with two or more heteroatoms. Adv.
Heterocycl. Chem. (2000) 76: 157-323.
Verma A and Saraf SK. 4-Thiazolidinone–A
biologically active scaffold. Eur. J. Med. Chem. (2008)
43: 897-905.
Tripathi AC, Gupta SJ, Fatima GN, Sonar PK, Verma
A and Saraf SK. 4-Thiazolidinones: the advances
continue. Eur. J. Med. Chem. (2014) 72: 52-77.
Sindhu J, Singh H, Khurana JM, Sharma C and Aneja
KR. Multicomponent domino process for the synthesis
of some novel 5-(arylidene)-3-((1-aryl-1H-1, 2,
3-triazol-4-yl) methyl)-thiazolidine-2, 4-diones using
PEG-400 as an efficient reaction medium and their
antimicrobial evaluation. Chin. Chem. Lett. (2015)
26: 50-4.
Malik S, Upadhyaya PK and Miglani S.
Thiazolidinediones: a plethro of biological load. Int. J.
PharmTech Res. (2011) 3: 62-75.
Day C. Thiazolidinediones: a new class of antidiabetic
drugs. Diabetes Res. (1999) 16: 179-92.
Fujiwara T, Yoshioka S, Yoshioka T, Ushiyama I and
Horikoshi H. Characterization of new oral antidiabetic
agent CS-045: studies in KK and ob/ob mice and
Zucker fatty rats. Diabetes (1988) 37: 1549-58.
Zebardast T, Zarghi A, Daraie B, Hedayati M and
Dadrass OG. Design and synthesis of 3-alkyl-2-
aryl-1, 3-thiazinan-4-one derivatives as selective
cyclooxygenase (COX-2) inhibitors. Bioorg. Med.
Chem. Lett. (2009) 19: 3162-5.
Zarghi A, Javid FS, Ghodsi R, Dadrass OG, Daraei
B and Hedayati M. Design, synthesis and biological
evaluation of new 5, 5-diarylhydantoin derivatives
as selective cyclooxygenase-2 inhibitors. Sci. Pharm.
(2011) 79: 449.
Schepetkin IA, Kirpotina LN, Khlebnikov AI, Hanks
(13)
(14)
(15)
(16)
(17)
(18)
(19)
(20)
(21)
(22)
(23)
(24)
(25)
(26)
(27)
Anti- inflammatory effects of Thiazolidinedione derivative on Raw 264.7 cells
1379
TS, Kochetkova I, Pascual DW, Jutila MA and Quinn
MT. Identification and characterization of a novel
class of c-Jun N-terminal kinase inhibitors. Mol.
Pharmacol. (2012) 81: 832-45.
Vazzana I, Terranova E, Mattioli F and Sparatore
F. Aromatic Schiff bases and 2, 3-disubstituted-1,
3-thiazolidin-4-one derivatives as antiinflammatory
agents. Arkivoc (2004) 5: 364-74.
Mohammadi A and Safarnejad M. Synthesis, structural
characterization and tautomeric properties of some
novel bis-azo dyes derived from 5-arylidene-2,
4-thiazolidinone. Spectrochim. Acta A Mol. Biomol.
Spectrosc. (2014) 126: 105-11.
Scudiero DA, Shoemaker RH, Paull KD, Monks
A, Tierney S, Nofziger TH, Currens MJ, Seniff D
and Boyd MR. Evaluation of a soluble tetrazolium/
formazan assay for cell growth and drug sensitivity in
culture using human and other tumor cell lines. Cancer
Res. (1988) 48: 4827-33.
Granger DL, Taintor RR and Boockvar KS.
Determination of nitrate and nitrite in biological
samples using bacterial nitrate reductase coupled with
the Griess reaction. Methods (1995) 7: 78-83.
Word JM, Lovell SC, Richardson JS and Richardson
DC. Asparagine and glutamine: using hydrogen atom
contacts in the choice of side-chain amide orientation.
J. Mol. Biol. (1999) 285: 1735-47.
Trott O and Olson AJ. Improving the speed and
accuracy of docking with a new scoring function,
efficient optimization, and multithreading. J. Comput.
Chemistry. (2010) 31: 455-61.
Sanner MF. Python: a programming language for
software integration and development. J. Mol. Graph.
Model. (1999) 17: 57-61.
Coleman WF and Arumainayagam CR. HyperChem 5
(by Hypercube, Inc.). J. Chem. Educ. (1998) 75: 416.
Wallace AC, Laskowski RA and Thornton JM.
LIGPLOT: a program to generate schematic diagrams
of protein-ligand interactions. Protein Eng. (1995) 8:
127-34.
Thiyam‐Holländer U, Aladedunye F, Logan A, Yang
H and Diehl BWK. Identification and quantification
of canolol and related sinapate precursors in Indian
mustard oils and Canadian mustard products. Eur. J.
Lipid. Sci. Tech. (2014) 116: 1664-74.
(28)
(29)
(30)
(31)
(32)
(33)
(34)
(35)
(36)
(37)
Namjoshi S and Benson HAE. Cyclic peptides as
potential therapeutic agents for skin disorders. Pept.
Sci. (2010) 94: 673-80.
Isomura Y, Yamaji Y, Yamada A, Watanabe Y, Suzuki
H, Kobayashi Y, Yoshida S, Watabe H, Hirata Y,
Yoshida H and Koike K. Irsogladine improves smallintestinal injuries in regular users of nonsteroidal
anti-inflammatory drugs. Gastrointest. Endosc. (2014)
80: 118-25.
Dinarello CA. Anti-inflammatory agents: present and
future. Cell (2010) 140: 935-50.
Thun MJ, Henley SJ and Patrono C. Nonsteroidal antiinflammatory drugs as anticancer agents: mechanistic,
pharmacologic, and clinical issues. J. Natl. Cancer
Inst. (2002) 94: 252-66.
Murakami A and Ohigashi H. Targeting NOX, INOS
and COX2 in inflammatory cells: Chemoprevention
using food phytochemicals. Int. J. Cancer (2007) 121:
2357-63.
Chiou W, Chen C and Lin J. Mechanisms of
suppression of inducible nitric oxide synthase (iNOS)
expression in RAW 264.7 cells by andrographolide.
Br. J. Pharmacol. (2000) 129: 1553-60.
Nussler AK and Billiar TR. Inflammation,
immunoregulation, and inducible nitric oxide synthase.
J. Leukoc. Biol. (1993) 54: 171-8.
Ma L, Pei H, Lei L, He L, Chen J, Liang X, Peng A,
Ye H, Xiang M and Chen L. Structural exploration,
synthesis and pharmacological evaluation of novel
5-benzylidenethiazolidine-2, 4-dione derivatives as
iNOS inhibitors against inflammatory diseases. Eur. J.
Med. Chem. (2015) 92: 178-90.
Ma L, Xie C, Ma Y, Liu J, Xiang M, Ye X, Zheng
H, Chen Zh, Xu Q, Chen T, Chen J, Yang J, Qiu
N, Wang G, Liang X, Peng A, Yang Sh, Wei Y and
Chen L. Synthesis and biological evaluation of novel
5-benzylidenethiazolidine-2, 4-dione derivatives for
the treatment of inflammatory diseases. J. Med. Chem.
(2011) 54: 2060-8.
Amin AR, Vyas P, Attur M, Leszczynska-Piziak J,
Patel IR, Weissmann G and Abramson SB. The mode
of action of aspirin-like drugs: effect on inducible
nitric oxide synthase. Proc. Natl. Acad. Sci. USA
(1995) 92: 7926-30