Indomethacin Affects the Inflammatory Response via Interaction with the RhoA-Actin Cytoskeleton in THP-1 Cells

Year 2023, Volume: 13 Issue: 3, 213 - 217, 28.12.2023

Ebru Hacıosmanoğlu Aldoğan , Fulya Dal Yöntem , Şeyma Bulut , Hande Yapislar , Başak Günçer , Muhammmet Bektaş

https://doi.org/10.26650/experimed.1364227

Abstract

Objective: Inflammation is a complex reaction present in numerous disorders. Indomethacin, a compound possessing an indoline core, is a Nonsteroidal Anti-Inflammatory Drug (NSAID) that is commonly prescribed for inflammation and pain. The actin network, plays a major role in cellular activities and it’s regulated by by Rho GTPases has important implications for cellular dynamics and orientation. In this research, we explore the effects of indomethacin on the inflammatory response as mediated via RhoA and pyrin inflammatory complexes using an inflammatory disease model with relation actin cytoskeleton.
Materials and Methods: This study used Western blotting to examine the impact of indomethacin on the assembly processes related to the pyrin inflammasome complex and the RhoA signaling pathway in Lipopolysaccharide-stimulated THP-1 cells. Actin-indomethacin interaction was analyzed by Differential Scanning Fluorimetry (DSF).
Results: We found that while the expression levels of pyrin decreased, phosphorylated-RhoA increased but overall RhoA levels did not change. The equilibrium dissociation constant (KD) for the G-actin-indomethacin complex was calculated to be 9.591± 1.608 ng/mL (R2 = 0.8582) using ∆Tm measurements of indomethacin by DSF.
Conclusion: Moreover, the effects of indomethacin on inflammation pathways may provide insight into the molecular mechanisms of pyrin inflammasome formation in various autoimmune diseases.

Keywords

Actin, differential scanning fluorimetry, GTPases, indomethacin, RhoA, Rho pyrin

Ethical Statement

Çalışma hücre kültürü çalışması olduğundan dolayı etik kurul onayına ihtiyaç yoktur

Supporting Institution

This study was funded by Scientific Research Projects Coordination Unit of Bezmialem Vakif University (Project number: 2.2019/17).

References

• 1. Ozen S. What’s new in autoinflammation? Pediatr Nephrol 2019; 34(12): 2449-56. google scholar
• 2. Samuels J, Ozen S. Familial Mediterranean fever and the other autoinflammatory syndromes: evaluation of the patient with recurrent fever. Curr Opin Rheumatol 2006; 18(1): 108-17. google scholar
• 3. Park YH, Wood G, Kastner DL, Chae JJ. Pyrin inflammasome activation and RhoA signaling in the autoinflammatory diseases FMF and HIDS. Nat Immunol 2016; 17(8): 914-21. google scholar
• 4. Waite AL, Schaner P, Hu C, Richards N, Balci-Peynircioglu B, Hong A, et al. Pyrin and ASC co-localize to cellular sites that are rich in polymerizing actin. Exp Biol Med (Maywood) 2009; 234(1): 40-52. google scholar
• 5. Kustermans G, Piette J, Legrand-Poels S. Actin-targeting natural compounds as tools to study the role of actin cytoskeleton in signal transduction. Biochem Pharmacol 2008; 76(11): 1310-22. google scholar
• 6. Rajakyla EK, Vartiainen MK. Rho, nuclear actin, and actin-binding proteins in the regulation of transcription and gene expression. Small GTPases 2014; 5: e27539. google scholar
• 7. Pantovic A, Bosnjak M, Arsikin K, Kosic M, Mandic M, Ristic B, et al. In vitro antiglioma action of indomethacin is mediated via AMP-activated protein kinase/mTOR complex 1 signalling pathway. Int J Biochem Cell Biol 2017; 83: 84-96. google scholar
• 8. Lu G, Pan Y, Kayoumu A, Zhang L, Yin T, Tong Z, et al. Indomethacin inhabits the NLRP3 inflammasome pathway and protects severe acute pancreatitis in mice. Biochem Biophys Res Commun 2017; 493(1): 827-32. google scholar
• 9. Alburquerque-Gonzalez B, Bernabe-Garcia A, Bernabe-Garcia M, Ruiz-Sanz J, Lopez-Calderon FF, Gonnelli L, et al. The FDA-approved antiviral raltegravir inhibits fascin1-dependent invasion of colorectal tumor cells in vitro and in vivo. Cancers (Basel) 2021; 13(4). google scholar
• 10. Vivoli M, Novak HR, Littlechild JA, Harmer NJ. Determination of protein-ligand interactions using differential scanning fluorimetry. J Vis Exp 2014; 91 : 51809. google scholar
• 11. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227(5259): 680-5. google scholar
• 12. Sen S, Haciosmanoglu E. Comparing the neuroprotective effects of telmisartan, perindopril, and nebivolol against lipopolysaccharide-induced injury in neuron-like cells. Cureus. 2022; 14(7): e27429. google scholar
• 13. Dominguez R, Holmes KC. Actin structure and function. Annu Rev Biophys 2011; 40: 169-86. google scholar
• 14. Misawa T, Takahama M, Kozaki T, Lee H, Zou J, Saitoh T, et al. Microtubule-driven spatial arrangement of mitochondria promotes activation of the NLRP3 inflammasome. Nat Immunol 2013; 14(5): 454-60. google scholar
• 15. Smith EC, Teixeira AM, Chen RC, Wang L, Gao Y, Hahn KL, et al. Induction of megakaryocyte differentiation drives nuclear accumulation and transcriptional function of MKL1 via actin polymerization and RhoA activation. Blood 2013; 121(7): 1094-101. google scholar
• 16. Ferrera P, Zepeda A, Arias C. Nonsteroidal anti-inflammatory drugs attenuate amyloid-beta protein-induced actin cytoskeletal reorganization through Rho signaling modulation. Cell Mol Neurobiol 2017; 37(7): 1311-8. google scholar
• 17. Riento K, Ridley AJ. Rocks: multifunctional kinases in cell behaviour. Nat Rev Mol Cell Biol 2003; 4(6): 446-56. google scholar