Extracellular Beta-nicotinamide Adenine Dinucleotide (beta-NAD) Promotes the Endothelial Cell Barrier Integrity Via PKA- and EPAC1/Rac1-dependent Actin Cytoskeleton Rearrangement

Overview

Journal J Cell Physiol

Specialties Cell Biology
Physiology

Date 2010 Jan 8

PMID 20054824

Citations 26

Authors

Nagavedi S Umapathy

Evgeny A Zemskov

Joyce Gonzales

Boris A Gorshkov

Supriya Sridhar

Trinad Chakraborty

Rudolf Lucas

Alexander D Verin

Affiliations

Soon will be listed here.

Abstract

Extracellular beta-NAD is known to elevate intracellular levels of calcium ions, inositol 1,4,5-trisphate and cAMP. Recently, beta-NAD was identified as an agonist for P2Y1 and P2Y11 purinergic receptors. Since beta-NAD can be released extracellularly from endothelial cells (EC), we have proposed its involvement in the regulation of EC permeability. Here we show, for the first time, that endothelial integrity can be enhanced in EC endogenously expressing beta-NAD-activated purinergic receptors upon beta-NAD stimulation. Our data demonstrate that extracellular beta-NAD increases the transendothelial electrical resistance (TER) of human pulmonary artery EC (HPAEC) monolayers in a concentration-dependent manner indicating endothelial barrier enhancement. Importantly, beta-NAD significantly attenuated thrombin-induced EC permeability as well as the barrier-compromising effects of Gram-negative and Gram-positive bacterial toxins representing the barrier-protective function of beta-NAD. Immunofluorescence microscopy reveals more pronounced staining of cell-cell junctional protein VE-cadherin at the cellular periphery signifying increased tightness of the cell-cell contacts after beta-NAD stimulation. Interestingly, inhibitory analysis (pharmacological antagonists and receptor sequence specific siRNAs) indicates the participation of both P2Y1 and P2Y11 receptors in beta-NAD-induced TER increase. beta-NAD-treatment attenuates the lipopolysaccharide (LPS)-induced phosphorylation of myosin light chain (MLC) indicating its involvement in barrier protection. Our studies also show the involvement of cAMP-dependent protein kinase A and EPAC1 pathways as well as small GTPase Rac1 in beta-NAD-induced EC barrier enhancement. With these results, we conclude that beta-NAD regulates the pulmonary EC barrier integrity via small GTPase Rac1- and MLCP- dependent signaling pathways.

Citing Articles

Extracellular purines in lung endothelial permeability and pulmonary diseases.

Gerasimovskaya E, Patil R, Davies A, Maloney M, Simon L, Mohamed B Front Physiol. 2024; 15():1450673.

PMID: 39234309 PMC: 11372795. DOI: 10.3389/fphys.2024.1450673.

Sirtuin 6 Regulates the Activation of the ATP/Purinergic Axis in Endothelial Cells.

Astigiano C, Piacente F, Laugieri M, Benzi A, Di Buduo C, Miguel C Int J Mol Sci. 2023; 24(7).

PMID: 37047732 PMC: 10095398. DOI: 10.3390/ijms24076759.

Regulation of Papillary Muscle Contractility by NAD and Ammonia Interplay: Contribution of Ion Channels and Exchangers.

Averin A, Konakov M, Pimenov O, Galimova M, Berezhnov A, Nenov M Membranes (Basel). 2022; 12(12).

PMID: 36557146 PMC: 9785361. DOI: 10.3390/membranes12121239.

Biological Properties of Vitamins of the B-Complex, Part 1: Vitamins B, B, B, and B.

Hrubsa M, Siatka T, Nejmanova I, Voprsalova M, Krcmova L, Matousova K Nutrients. 2022; 14(3).

PMID: 35276844 PMC: 8839250. DOI: 10.3390/nu14030484.

P2Y Purinergic Receptors, Endothelial Dysfunction, and Cardiovascular Diseases.

Strassheim D, Verin A, Batori R, Nijmeh H, Burns N, Kovacs-Kasa A Int J Mol Sci. 2020; 21(18).

PMID: 32962005 PMC: 7555413. DOI: 10.3390/ijms21186855.

References

Koh H, Tasaka S, Hasegawa N, Yamada W, Shimizu M, Nakamura M . Protective role of vascular endothelial growth factor in endotoxin-induced acute lung injury in mice. Respir Res. 2007; 8:60. PMC: 2042500. DOI: 10.1186/1465-9921-8-60. View

Moldobaeva A, Welsh-Servinsky L, Shimoda L, Stephens R, Verin A, Tuder R . Role of protein kinase G in barrier-protective effects of cGMP in human pulmonary artery endothelial cells. Am J Physiol Lung Cell Mol Physiol. 2005; 290(5):L919-30. DOI: 10.1152/ajplung.00434.2005. View

Mutafova-Yambolieva V, Hwang S, Hao X, Chen H, Zhu M, Wood J . Beta-nicotinamide adenine dinucleotide is an inhibitory neurotransmitter in visceral smooth muscle. Proc Natl Acad Sci U S A. 2007; 104(41):16359-64. PMC: 2042211. DOI: 10.1073/pnas.0705510104. View

Garcia J, Schaphorst K . Regulation of endothelial cell gap formation and paracellular permeability. J Investig Med. 1995; 43(2):117-26. View

Dudek S, Camp S, Chiang E, Singleton P, Usatyuk P, Zhao Y . Pulmonary endothelial cell barrier enhancement by FTY720 does not require the S1P1 receptor. Cell Signal. 2007; 19(8):1754-64. PMC: 2682440. DOI: 10.1016/j.cellsig.2007.03.011. View

Knezevic N, Roy A, Timblin B, Konstantoulaki M, Sharma T, Malik A . GDI-1 phosphorylation switch at serine 96 induces RhoA activation and increased endothelial permeability. Mol Cell Biol. 2007; 27(18):6323-33. PMC: 2099605. DOI: 10.1128/MCB.00523-07. View

Verin A, Birukova A, Wang P, Liu F, Becker P, Birukov K . Microtubule disassembly increases endothelial cell barrier dysfunction: role of MLC phosphorylation. Am J Physiol Lung Cell Mol Physiol. 2001; 281(3):L565-74. DOI: 10.1152/ajplung.2001.281.3.L565. View

Kolosova I, Mirzapoiazova T, Adyshev D, Usatyuk P, Romer L, Jacobson J . Signaling pathways involved in adenosine triphosphate-induced endothelial cell barrier enhancement. Circ Res. 2005; 97(2):115-24. DOI: 10.1161/01.RES.0000175561.55761.69. View

Krebs C, Adriouch S, Braasch F, Koestner W, Leiter E, Seman M . CD38 controls ADP-ribosyltransferase-2-catalyzed ADP-ribosylation of T cell surface proteins. J Immunol. 2005; 174(6):3298-305. DOI: 10.4049/jimmunol.174.6.3298. View

10.

Fukuhara S, Sakurai A, Sano H, Yamagishi A, Somekawa S, Takakura N . Cyclic AMP potentiates vascular endothelial cadherin-mediated cell-cell contact to enhance endothelial barrier function through an Epac-Rap1 signaling pathway. Mol Cell Biol. 2004; 25(1):136-46. PMC: 538793. DOI: 10.1128/MCB.25.1.136-146.2005. View

11.

Rebecchi M, Pentyala S . Structure, function, and control of phosphoinositide-specific phospholipase C. Physiol Rev. 2000; 80(4):1291-335. DOI: 10.1152/physrev.2000.80.4.1291. View

12.

Jaffe A, Hall A . Rho GTPases in transformation and metastasis. Adv Cancer Res. 2002; 84:57-80. DOI: 10.1016/s0065-230x(02)84003-9. View

13.

Watanabe T, Wang S, Noritake J, Sato K, Fukata M, Takefuji M . Interaction with IQGAP1 links APC to Rac1, Cdc42, and actin filaments during cell polarization and migration. Dev Cell. 2004; 7(6):871-83. DOI: 10.1016/j.devcel.2004.10.017. View

14.

Bai J, Deng W, Zhang J, Xu S, Zhang D . [Protective effect of myosin light-chain kinase inhibitor on acute lung injury]. Zhongguo Wei Zhong Bing Ji Jiu Yi Xue. 2009; 21(4):215-8. View

15.

Lum H, Malik A . Mechanisms of increased endothelial permeability. Can J Physiol Pharmacol. 1996; 74(7):787-800. DOI: 10.1139/y96-081. View

16.

Birukova A, Zagranichnaya T, Fu P, Alekseeva E, Chen W, Jacobson J . Prostaglandins PGE(2) and PGI(2) promote endothelial barrier enhancement via PKA- and Epac1/Rap1-dependent Rac activation. Exp Cell Res. 2007; 313(11):2504-20. PMC: 1974901. DOI: 10.1016/j.yexcr.2007.03.036. View

17.

Scheuplein F, Schwarz N, Adriouch S, Krebs C, Bannas P, Rissiek B . NAD+ and ATP released from injured cells induce P2X7-dependent shedding of CD62L and externalization of phosphatidylserine by murine T cells. J Immunol. 2009; 182(5):2898-908. DOI: 10.4049/jimmunol.0801711. View

18.

Birukova A, Smurova K, Birukov K, Usatyuk P, Liu F, Kaibuchi K . Microtubule disassembly induces cytoskeletal remodeling and lung vascular barrier dysfunction: role of Rho-dependent mechanisms. J Cell Physiol. 2004; 201(1):55-70. DOI: 10.1002/jcp.20055. View

19.

Wojciak-Stothard B, Tsang L, Paleolog E, Hall S, Haworth S . Rac1 and RhoA as regulators of endothelial phenotype and barrier function in hypoxia-induced neonatal pulmonary hypertension. Am J Physiol Lung Cell Mol Physiol. 2006; 290(6):L1173-82. DOI: 10.1152/ajplung.00309.2005. View

20.

Ware L, Matthay M . The acute respiratory distress syndrome. N Engl J Med. 2000; 342(18):1334-49. DOI: 10.1056/NEJM200005043421806. View