Top

Published in:

Open Access 01-12-2023 | Research

Mechanisms of glabridin inhibition of integrin α_IIbβ₃ inside-out signals and NF-κB activation in human platelets

Authors: Wei-Chieh Huang, Thanasekaran Jayakumar, Joen-Rong Sheu, Chih-Wei Hsia, Chih-Hsuan Hsia, Ting-Lin Yen, Chao-Chien Chang

Published in: Chinese Medicine | Issue 1/2023

Abstract

Background

Platelets play a crucial role in cardiovascular diseases (CVDs) and are activated by endogenous agonists like collagen. These agonists initiate signal transduction through specific platelet receptors, resulting in platelet aggregation. Glabridin, a prenylated isoflavonoid found in licorice root, is known for its significance in metabolic abnormalities. Glabridin has been observed to inhibit collagen-induced platelet aggregation, but the precise mechanisms, specifically concerning NF-κB activation and integrin α_IIbβ₃ signaling, are not yet fully understood.

Methods

In this study, platelet suspensions were prepared from healthy human blood donors, and the aggregation ability was observed using a lumi-aggregometer. The inhibitory mechanisms of glabridin in human platelets were evaluated through immunoblotting and confocal microscopy. The anti-thrombotic effects of glabridin were assessed by histological analysis of lung sections in acute pulmonary thromboembolism and by examining fluorescein-induced platelet plug formation in mesenteric microvessels in mice.

Results

Glabridin inhibited integrin α_IIbβ₃ inside-out signals such as Lyn, Fyn, Syk, and integrin β₃ activation and NF-κB-mediated signal events, with similar potency to classical inhibitors BAY11-7082 and Ro106-9920. Glabridin and BAY11-7082 inhibited IKK, IκBα, and p65 phosphorylation and reversed IκBα degradation, while Ro106-9920 only reduced p65 phosphorylation and reversed IκBα degradation. BAY11-7082 reduced Lyn, Fyn, Syk, integrin β₃, phospholipase Cγ2 and protein kinase C activation. Glabridin reduced platelet plug formation in mesenteric microvessels and occluded vessels in thromboembolic lungs of mice.

Conclusion

Our study revealed a new pathway for activating integrin α_IIbβ₃ inside-out signals and NF-κB, which contributes to the antiplatelet aggregation effect of glabridin. Glabridin could be a valuable prophylactic or clinical treatment option for CVDs.

Available only for authorised users

Nagareddy P, Smyth SS. Inflammation and thrombosis in cardiovascular disease. Curr Opin Hematol. 2013;20:457–63. https://doi.org/10.1097/MOH.0b013e328364219d.CrossRefPubMedPubMedCentral

Khodadi E. Platelet function in cardiovascular disease: activation of molecules and activation by molecules. Cardiovasc Toxicol. 2020;20:1–10. https://doi.org/10.1007/s12012-019-09555-4.CrossRefPubMed

Sheu JR, Yen MH, Hung WC, Lee YM, Su CH, Huang TF. Triflavin inhibits platelet-induced vasoconstriction in de-endothelialized aorta. Arterioscler Thromb Vasc Biol. 1997;17:3461–8. https://doi.org/10.1161/01.atv.17.12.3461.CrossRefPubMed

Payrastre B, Missy K, Trumel C, Bodin S, Plantavid M, Chap H. The integrin alpha IIb/beta 3 in human platelet signal transduction. Biochem Pharmacol. 2000;60:1069–74. https://doi.org/10.1016/s0006-2952(00)00417-2.CrossRefPubMed

Pikarsky E, Porat RM, Stein I, Abramovitch R, Amit S, Kasem S, Gutkovich-Pyest E, Urieli-Shoval S, Galun E, Ben-Neriah Y. NF-kappaB functions as a tumour promoter in inflammation-associated cancer. Nature. 2004;431:461–6. https://doi.org/10.1038/nature02924.CrossRefPubMed

Hsia CW, Huang WC, Yang CH, Hsia CH, Jayakumar T, Bhavan PS, Sheu JR. Chiou KR comparison of the potency of pterostilbene with NF-κB inhibitors in platelet activation: mutual activation by Akt-NF-κB signaling in human platelets. Appl Sci. 2021;11:6149. https://doi.org/10.3390/app11136149.CrossRef

Wang X, Zhang H, Chen L, Shan L, Fan G, Gao X. Liquorice, a unique “guide drug” of traditional chinese medicine: a review of its role in drug interactions. J Ethnopharmacol. 2013;150:781–90. https://doi.org/10.1016/j.jep.2013.09.055.CrossRefPubMed

Li C, Li T, Zhu M, Lai J, Wu Z. Pharmacological properties of glabridin (a flavonoid extracted from licorice): a comprehensive review. J Funct Foods. 2021;85:104638. https://doi.org/10.1016/j.jff.2021.104638.CrossRef

Carmeli E, Harpaz Y, Kogan NN, Fogelman Y. The effect of an endogenous antioxidant glabridin on oxidized LDL. J Basic Clin Physiol Pharmacol. 2008;9:49–63. https://doi.org/10.1515/jbcpp.2008.19.1.49.CrossRef

10.

Kang MR, Park KH, Oh SJ, Yun J, Lee CW, Lee MY, Han S, Kang JS. Cardiovascular protective effect of glabridin: implications in LDL oxidation and inflammation. Int Immunopharmacol. 2015;29:914–8. https://doi.org/10.1016/j.intimp.2015.10.020.CrossRefPubMed

11.

Lien L, Lin K, Huang L, Tseng M, Chiu H, Chen R, Lu W. Licochalcone a prevents platelet activation and thrombus formation through the inhibition of PLCγ2-PKC, akt, and MAPK pathways. Int J Mol Sci. 2017;18:1500. https://doi.org/10.3390/ijms18071500.CrossRefPubMedPubMedCentral

12.

Chung C, Chen J, Huang W, Sheu J, Hsia C, Jayakumar T, Hsia C, Chiou K, Hou S. Glabridin, a bioactive flavonoid from licorice, effectively inhibits platelet activation in humans and mice. Int J Mol Sci. 2022;23:11372. https://doi.org/10.3390/ijms231911372.CrossRefPubMedPubMedCentral

13.

Chen WF, Lee JJ, Chang CC, Lin KH, Wang SH, Sheu JR. Platelet protease-activated receptor (PAR)4, but not PAR1, associated with neutral sphingomyelinase responsible for thrombin-stimulated ceramide-NF-κB signaling in human platelets. Haematologica. 2013;98:793–801. https://doi.org/10.3324/haematol.2012.072553.CrossRefPubMedPubMedCentral

14.

Sheu JR, Lee CR, Lin CH, Hsiao G, Ko WC, Chen YC, Yen MH. Mechanisms involved in the antiplatelet activity of Staphylococcus aureus lipoteichoic acid in human platelets. Thromb Haemost. 2000;83:777–84. https://doi.org/10.1055/s-0037-1613907.CrossRefPubMed

15.

Crosby D, Poole AW. Physical and functional interaction between protein kinase C delta and fyn tyrosine kinase in human platelets. J Biol Chem. 2003;278:24533–41. https://doi.org/10.1074/jbc.M301847200.CrossRefPubMed

16.

Sheu JR, Hung WC, Wu CH, Lee YM, Yen MH. Antithrombotic effect of rutaecarpine, an alkaloid isolated from Evodia rutaecarpa, on platelet plug formation in in vivo experiments. Br J Haematol. 2000;110:110–5. https://doi.org/10.1046/j.1365-2141.2000.01953.x.CrossRefPubMed

17.

Hsiao G, Lin KH, Chang Y, Chen TL, Tzu NH, Chou DS, Sheu JR. Protective mechanisms of inosine in platelet activation and cerebral ischemic damage. Arterioscler Thromb Vasc Biol. 2005;25:1998–2004.CrossRefPubMed

18.

Varga-Szabo D, Braun A, Nieswandt B. Calcium signaling in platelets. J Thromb Haemost. 2009;7:1057–66. https://doi.org/10.1111/j.1538-7836.2009.03455.x.CrossRefPubMed

19.

Lin Y, Bai L, Chen W, Xu S. The NF-kappaB activation pathways, emerging molecular targets for cancer prevention and therapy. Expert Opin Ther Targets. 2010;14:45–55. https://doi.org/10.1517/14728220903431069.CrossRefPubMedPubMedCentral

20.

Hosseinzadeh H, Nassiri-Asl M. Pharmacological effects of Glycyrrhiza spp. and its bioactive constituents: update and review. Phytother Res. 2015;29:1868–86. https://doi.org/10.1002/ptr.5487.CrossRefPubMed

21.

Phillips DR, Nannizzi-Alaimo L, Prasad KS. Beta3 tyrosine phosphorylation in alphaIIbbeta3 (platelet membrane GP IIb-IIIa) outside-in integrin signaling. Thromb Haemost. 2001;86:246–58. https://doi.org/10.1055/s-0037-1616222.CrossRefPubMed

22.

Sheu JR, Hung WC, Wu CH, Ma MC, Kan YC, Lin CH, Lin MS, Luk HN, Yen MH. Reduction in lipopolysaccharide-induced thrombocytopenia by triflavin in a rat model of septicemia. Circulation. 1999;99:3056–62. https://doi.org/10.1161/01.cir.99.23.3056.CrossRefPubMed

23.

Huang TF, Sheu JR, Teng CM. A potent antiplatelet peptide, triflavin, from Trimeresurus flavoviridis snake venom. Biochem J. 1991;277:351–7. https://doi.org/10.1042/bj2770351.CrossRefPubMedPubMedCentral

24.

Shattil SJ, Newman PJ. Integrins: dynamic scaffolds for adhesion and signaling in platelets. Blood. 2004;104:1606–15. https://doi.org/10.1182/blood-2004-04-1257.CrossRefPubMed

25.

Estevez B, Du X. New concepts and mechanisms of platelet activation signaling. Physiol (Bethesda). 2017;32:162–77. https://doi.org/10.1152/physiol.00020.2016.CrossRef

26.

Law DA, DeGuzman FR, Heiser P, Ministri-Madrid K, Killeen N, Phillips DR. Integrin cytoplasmic tyrosine motif is required for outside-in αIIbβ3 signaling and platelet function. Nature. 1999;401:808–11. https://doi.org/10.1038/44599.CrossRefPubMed

27.

Ghosh S, Hayden MS. New regulators of NF-kappaB in inflammation. Nat Rev Immunol. 2008;8:837–48. https://doi.org/10.1038/nri2423.CrossRefPubMed

28.

Monaco C, Andreakos E, Kiriakidis S, Mauri C, Bicknell C, Foxwell B, Cheshire N, Paleolog E, Feldmann M. Canonical pathway of nuclear factor kappa B activation selectively regulates proinflammatory and prothrombotic responses in human atherosclerosis. Proc Natl Acad Sci USA. 2004;101:5634–9. https://doi.org/10.1073/pnas.0401060101.CrossRefPubMedPubMedCentral

29.

Malaver E, Romaniuk MA, D’Atri LP, Pozner RG, Negrotto S, Benzadon R, Schattner M. NF-kappaB inhibitors impair platelet activation responses. J Thromb Haemost. 2009;7:1333–43. https://doi.org/10.1111/j.1538-7836.2009.03492.x.CrossRefPubMed

Title: Mechanisms of glabridin inhibition of integrin αIIbβ3 inside-out signals and NF-κB activation in human platelets
Authors: Wei-Chieh Huang
Thanasekaran Jayakumar
Joen-Rong Sheu
Chih-Wei Hsia
Chih-Hsuan Hsia
Ting-Lin Yen
Chao-Chien Chang
Publication date: 01-12-2023
Publisher: BioMed Central
Published in: Chinese Medicine / Issue 1/2023
Electronic ISSN: 1749-8546
DOI: https://doi.org/10.1186/s13020-023-00779-9

EASD 2024

Springer Medicine

Mechanisms of glabridin inhibition of integrin α_IIbβ₃ inside-out signals and NF-κB activation in human platelets

Abstract

Background

Methods

Results

Conclusion

EASD 2024

Springer Medicine

Abstract

Background

Methods

Results

Conclusion

Please log in to get access to this content

Other articles of this Issue 1/2023

Rosmarinic acid treatment protects against lethal H1N1 virus-mediated inflammation and lung injury by promoting activation of the h-PGDS-PGD2-HO-1 signal axis

Pseudo-allergic reactions induced by Chinese medicine injections: a review

Research progress on antitumor mechanisms and molecular targets of Inula sesquiterpene lactones

Coix seed oil alleviates synovial angiogenesis through suppressing HIF-1α/VEGF-A signaling pathways via SIRT1 in collagen-induced arthritis rats

Correction: Saikosaponin D exerts antidepressant effect by regulating Homer1-mGluR5 and mTOR signaling in a rat model of chronic unpredictable mild stress

Kai-Xin-San protects against mitochondrial dysfunction in Alzheimer’s disease through SIRT3/NLRP3 pathway