Endothelial cell senescence is associated with disrupted cell-cell junctions and increased monolayer permeability

Journal of Angiogenesis Research

Jan Andries Post¹  Elsa Regan-Klapisz¹  Miriam Langelaar-Makkinje¹  Liesbeth H P Hekking¹  Vincent J D Krouwer¹ 

[1] Department of Biomolecular Imaging, Faculty of Science, Institute of Biomembranes, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands

关键词: cPLA₂α;    Endothelial permeability;    Endothelial junctions;    Atherosclerosis;    Senescence;   

Others  :  801975
DOI  :  10.1186/2045-824X-4-12

 received in 2012-03-30, accepted in 2012-08-19,  发布年份 2012

【 摘 要 】

Background

Cellular senescence is associated with cellular dysfunction and has been shown to occur in vivo in age-related cardiovascular diseases such as atherosclerosis. Atherogenesis is accompanied by intimal accumulation of LDL and increased extravasation of monocytes towards accumulated and oxidized LDL, suggesting an affected barrier function of vascular endothelial cells. Our objective was to study the effect of cellular senescence on the barrier function of non-senescent endothelial cells.

Methods

Human umbilical vein endothelial cells were cultured until senescence. Senescent cells were compared with non-senescent cells and with co-cultures of non-senescent and senescent cells. Adherens junctions and tight junctions were studied. To assess the barrier function of various monolayers, assays to measure permeability for Lucifer Yellow (LY) and horseradish peroxidase (PO) were performed.

Results

The barrier function of monolayers comprising of senescent cells was compromised and coincided with a change in the distribution of junction proteins and a down-regulation of occludin and claudin-5 expression. Furthermore, a decreased expression of occludin and claudin-5 was observed in co-cultures of non-senescent and senescent cells, not only between senescent cells but also along the entire periphery of non-senescent cells lining a senescent cell.

Conclusions

Our findings show that the presence of senescent endothelial cells in a non-senescent monolayer disrupts tight junction morphology of surrounding young cells and increases the permeability of the monolayer for LY and PO.

【 授权许可】

   
2012 Krouwer et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Eman MR, Regan-Klapisz E, Pinkse MW, Koop IM, Haverkamp J, Heck AJ, Verkleij AJ, Post JA: Protein expression dynamics during replicative senescence of endothelial cells studied by 2-D difference in-gel electrophoresis. Electrophoresis 2006, 27:1669-1682.
• [2]Hampel B, Fortschegger K, Ressler S, Chang MW, Unterluggauer H, Breitwieser A, Sommergruber W, Fitzky B, Lepperdinger G, Jansen-Durr P, Voglauer R, Grillari J: Increased expression of extracellular proteins as a hallmark of human endothelial cell in vitro senescence. Exp Gerontol 2006, 41:474-481.
• [3]Campisi J, d’Adda di Fagagna F: Cellular senescence: when bad things happen to good cells. Nat Rev Mol Cell Biol 2007, 8:729-740.
• [4]Collado M, Blasco MA, Serrano M: Cellular senescence in cancer and aging. Cell 2007, 130:223-233.
• [5]Unterluggauer H, Hutter E, Voglauer R, Grillari J, Voth M, Bereiter-Hahn J, Jansen-Durr P, Jendrach M: Identification of cultivation-independent markers of human endothelial cell senescence in vitro. Biogerontology 2007, 8:383-397.
• [6]Coppe JP, Desprez PY, Krtolica A, Campisi J: The senescence-associated secretory phenotype: the dark side of tumor suppression. Annu. Rev. Pathol. 2010, 5:99-118.
• [7]Mun GI, Boo YC: Identification of CD44 as a senescence-induced cell adhesion gene responsible for the enhanced monocyte recruitment to senescent endothelial cells. Am J Physiol Heart Circ Physiol 2010, 298:H2102-H2111.
• [8]Erusalimsky JD: Vascular endothelial senescence: from mechanisms to pathophysiology. J Appl Physiol 2009, 106:326-332.
• [9]Boisen L, Drasbek KR, Pedersen AS, Kristensen P: Evaluation of endothelial cell culture as a model system of vascular ageing. Exp Gerontol 2010, 45:779-787.
• [10]Vasile E, Tomita Y, Brown LF, Kocher O, Dvorak HF: Differential expression of thymosin beta-10 by early passage and senescent vascular endothelium is modulated by VPF/VEGF: evidence for senescent endothelial cells in vivo at sites of atherosclerosis. FASEB J 2001, 15:458-466.
• [11]Minamino T, Miyauchi H, Yoshida T, Ishida Y, Yoshida H, Komuro I: Endothelial cell senescence in human atherosclerosis: role of telomere in endothelial dysfunction. Circulation 2002, 105:1541-1544.
• [12]Bazzoni G, Dejana E: Endothelial cell-to-cell junctions: molecular organization and role in vascular homeostasis. Physiol Rev 2004, 84:869-901.
• [13]van Hinsbergh VW, van Nieuw Amerongen GP: Endothelial hyperpermeability in vascular leakage. Vascul Pharmacol 2002, 39(4–5):171-172.
• [14]Kim TW, Kim HJ, Lee C, Kim HY, Baek SH, Kim JH, Kwon KS, Kim JR: Identification of replicative senescence-associated genes in human umbilical vein endothelial cells by an annealing control primer system. Exp Gerontol 2008, 43:286-295.
• [15]Collins C, Tzima E: Hemodynamic forces in endothelial dysfu8nction and vascular ageing. Exp Gerontol 2011, 46(2–3):185-188.
• [16]Maier JA, Statuto M, Ragnotti G: Senescence stimulates U937-endothelial cell interactions. Exp. Cell Res. 1993, 208:270-274.
• [17]Op den Buijs J, Musters M, Verrips T, Post JA, Braam B, van Riel N: Mathematical modeling of vascular endothelial layer maintenance: the role of endothelial cell division, progenitor cell homing, and telomere shortening. Am J Physiol Heart Circ Physiol 2004, 287:H2651-H2658.
• [18]Libby P: Inflammation in atherosclerosis. Nature 2002, 420:868-874.
• [19]Lusis AJ: Atherosclerosis. Nature 2000, 407:233-241.
• [20]Fenton M, Barker S, Kurz DJ, Erusalimsky JD: Cellular senescence after single and repeated balloon catheter denudations of rabbit carotid arteries. Arterioscler Thromb Vasc Biol 2001, 21:220-226.
• [21]Ogami M, Ikura Y, Ohsawa M, Matsuo T, Kayo S, Yoshimi N, Hai E, Shirai N, Ehara S, Komatsu R, Naruko T, Ueda M: Telomere shortening in human coronary artery diseases. Arterioscler Thromb Vasc Biol 2004, 24:546-550.
• [22]Huang W, Rha GB, Chen L, Seelbach MJ, Zhang B, Andras IE, Bruemmer D, Hennig B, Toborek M: Inhibition of telomerase activity alters tight junction protein expression and induces transendothelial migration of HIV-1-infected cells. Am J Physiol Heart Circ Physiol 2010, 298:H1136-H1145.
• [23]Venkatesh D, Fredette N, Rostama B, Tang Y, Vary CP, Liaw L, Urs S: RhoA-mediated signaling in Notch-induced senescence-like growth arrest and endothelial barrier dysfunction. Arterioscler Thromb Vasc Biol 2011, 31:876-882.
• [24]Lee MY, Wang Y, Vanhoutte PM: Senescence of cultured porcine coronary arterial endothelial cells is associated with accelerated oxidative stress and activation of NFkB. J Vasc Res 2010, 47:287-298.
• [25]Regan-Klapisz E, Krouwer V, Langelaar-Makkinje M, Nallan L, Gelb M, Gerritsen H, Verkleij AJ, Post JA: Golgi-associated cPLA2alpha regulates endothelial cell-cell junction integrity by controlling the trafficking of transmembrane junction proteins. Mol. Biol. Cell 2009, 20:4225-4234.
• [26]Jiménez N, Vocking K, van Donselaar EG, Humbel BM, Post JA, Verkleij AJ: Tannic acid-mediated osmium impregnation after freeze-substitution: a strategy to enhance membrane contrast for electron tomography. J Struct Biol 2009, 166(1):103-106.
• [27]Jiménez N, Post JA: A novel approach for intracellular 3D immuno-labeling for electron-tomography. Traffic 2012.
• [28]Klapisz E, Sorokina I, Lemeer S, Pijnenburg M, Verkleij AJ, van Bergen en Henegouwen PM: A ubiquitin-interacting motif (UIM) is essential for Eps15 and Eps15R ubiquitination. J Biol Chem 2002, 277:30746-30753.
• [29]Taddei A, Giampietro C, Conti A, Orsenigo F, Breviario F, Pirazzoli V, Potente M, Daly C, Dimmeler S, Dejana E: Endothelial adherens junctions control tight junctions by VE-cadherin-mediated upregulation of claudin-5. Nat. Cell Biol. 2008, 10:923-934.
• [30]Campisi J: Senescent cells, tumor suppression, and organismal aging: good citizens, bad neighbors. Cell 2005, 120:513-522.
• [31]Cancel LM, Tarbell JM: The role of apoptosis in LDL transport through cultured endothelial cell monolayers. Atherosclerosis 2010, 208:335-341.
• [32]Chiu JJ, Usami S, Chien S: Vascular endothelial responses to altered shear stress: pathologic implications for atherosclerosis. Ann. Med. 2009, 41:19-28.
• [33]Chen Z, Peng IC, Cui X, Li YS, Chien S, Shyy JY: Shear stress, SIRT1, and vascular homeostasis. Proc Natl Acad Sci U S A 2010, 107:10268-10273.
• [34]Roy Chowdhury SK, Sangle GV, Xie X, Stelmack GL, Halayko AJ, Shen GX: Effects of extensively oxidized low-density lipoprotein on mitochondrial function and reactive oxygen species in porcine aortic endothelial cells. Am J Physiol Endocrinol Metab 2010, 298:E89-E98.