Upon inflammation, monocyte-derived macrophages (MΦ) infiltrate blood vessels to regulate several processes involved in vascular pathophysiology. However, little is known about the mediators involved. Macrophage polarization is crucial for a fast and efficient initial response (GM-MΦ) and a good resolution (M-MΦ) of the inflammatory process. The functional activity of polarized MΦ is exerted mainly through their secretome, which can target other cell types, including endothelial cells. Endoglin (CD105) is a cell surface receptor expressed by endothelial cells and MΦ that is markedly upregulated in inflammation and critically involved in angiogenesis. In addition, a soluble form of endoglin with anti-angiogenic activity has been described in inflammation-associated pathologies. The aim of this work was to identify components of the MΦ secretome involved in the shedding of soluble endoglin. We find that the GM-MΦ secretome contains metalloprotease 12 (MMP-12), a GM-MΦ specific marker that may account for the anti-angiogenic activity of the GM-MΦ secretome. Cell surface endoglin is present in both GM-MΦ and M-MΦ, but soluble endoglin is only detected in GM-MΦ culture supernatants. Moreover, MMP-12 is responsible for the shedding of soluble endoglin in vitro and in vivo by targeting membrane-bound endoglin in both MΦ and endothelial cells. These data demonstrate a direct correlation between GM-MΦ polarization, MMP-12, and soluble endoglin expression and function. By targeting endothelial cells, MMP-12 may represent a novel mediator involved in vascular homeostasis.
- Klíčová slova
- MMP-12, endoglin, endothelial cells, inflammation, macrophages, monocytes,
- MeSH
- biologické modely MeSH
- endoglin genetika metabolismus MeSH
- endoteliální buňky metabolismus MeSH
- exprese genu MeSH
- faktor stimulující granulocyto-makrofágové kolonie metabolismus MeSH
- faktor stimulující kolonie makrofágů metabolismus MeSH
- kultivované buňky MeSH
- lidé MeSH
- makrofágy imunologie metabolismus MeSH
- matrixová metaloproteinasa 12 metabolismus MeSH
- mediátory zánětu metabolismus MeSH
- modely nemocí na zvířatech MeSH
- myši MeSH
- náchylnost k nemoci MeSH
- zánět etiologie metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- endoglin MeSH
- faktor stimulující granulocyto-makrofágové kolonie MeSH
- faktor stimulující kolonie makrofágů MeSH
- matrixová metaloproteinasa 12 MeSH
- mediátory zánětu MeSH
Our objective was to investigate the effect of cholesterol [hypercholesterolemia and 7-ketocholesterol (7K)] on endoglin (Eng) expression and regulation with respect to endothelial or vascular dysfunction in vivo and in vitro. In vivo experiments were performed in 2-mo-old atherosclerosis-prone apolipoprotein E-deficient/LDL receptor-deficient (ApoE-/-/LDLR-/-) female mice and their wild-type C57BL/6J littermates. In in vitro experiments, human aortic endothelial cells (HAECs) were treated with 7K. ApoE-/-/LDLR-/- mice developed hypercholesterolemia accompanied by increased circulating levels of P-selectin and Eng and a disruption of NO metabolism. Functional analysis of the aorta demonstrated impaired vascular reactivity, and Western blot analysis revealed down-regulation of membrane Eng/Smad2/3/eNOS signaling in ApoE-/-/LDLR-/- mice. 7K increased Eng expression via Krüppel-like factor 6 (KLF6), liver X nuclear receptor, and NF-κB in HAECs. 7K-induced Eng expression was prevented by the treatment with 2-hydroxypropyl-β-cyclodextrin; 8-{[5-chloro-2-(4-methylpiperazin-1-yl) pyridine-4-carbonyl] amino}-1-(4-fluorophenyl)-4, 5-dihydrobenzo[g]indazole-3-carboxamide; or by KLF6 silencing. 7K induced increased adhesion and transmigration of monocytic human leukemia promonocytic cell line cells and was prevented by Eng silencing. We concluded that hypercholesterolemia altered Eng expression and signaling, followed by endothelial or vascular dysfunction before formation of atherosclerotic lesions in ApoE-/-/LDLR-/- mice. By contrast, 7K increased Eng expression and induced inflammation in HAECs, which was followed by an increased adhesion and transmigration of monocytes via endothelium, which was prevented by Eng inhibition. Thus, we propose a relevant role for Eng in endothelial or vascular dysfunction or inflammation when exposed to cholesterol.-Vicen, M., Vitverova, B., Havelek, R., Blazickova, K., Machacek, M., Rathouska, J., Najmanová, I., Dolezelova, E., Prasnicka, A., Sternak, M., Bernabeu, C., Nachtigal, P. Regulation and role of endoglin in cholesterol-induced endothelial and vascular dysfunction in vivo and in vitro.
- Klíčová slova
- endothelial cells, hypercholesterolemia, mice, oxysterols,
- MeSH
- aorta cytologie metabolismus patologie MeSH
- apolipoproteiny E genetika MeSH
- aterosklerotický plát etiologie genetika metabolismus MeSH
- beta-cyklodextriny farmakologie MeSH
- cévní endotel účinky léků metabolismus patologie MeSH
- cholesterol metabolismus MeSH
- endoglin genetika metabolismus MeSH
- hypercholesterolemie komplikace genetika metabolismus MeSH
- indazoly farmakologie MeSH
- KLF6 metabolismus MeSH
- kultivované buňky MeSH
- kyseliny isonikotinové farmakologie MeSH
- lidé MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- NF-kappa B metabolismus MeSH
- oxid dusnatý metabolismus MeSH
- P-selektin metabolismus MeSH
- proteiny Smad metabolismus MeSH
- receptory LDL genetika MeSH
- synthasa oxidu dusnatého, typ III metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- apolipoproteiny E MeSH
- beta-cyklodextriny MeSH
- cholesterol MeSH
- endoglin MeSH
- indazoly MeSH
- Klf6 protein, mouse MeSH Prohlížeč
- KLF6 MeSH
- kyseliny isonikotinové MeSH
- NF-kappa B MeSH
- oxid dusnatý MeSH
- P-selektin MeSH
- PHA 408 MeSH Prohlížeč
- proteiny Smad MeSH
- receptory LDL MeSH
- synthasa oxidu dusnatého, typ III MeSH
BACKGROUND AND AIMS: Increased plasma levels of soluble endoglin (sEng) were detected in patients with endothelial dysfunction-related disorders and hypercholesterolemia. In this study, we hypothesized that high levels of sEng accompanied by mild hypercholesterolemia could aggravate endothelial and vessel wall dysfunction and affect endoglin/eNOS signaling in mouse aorta. METHODS: Three-month-old female transgenic mice on CBAxC57BL/6J background, with high levels of sEng (Sol-Eng+high HFD), and their littermates with low levels of sEng (Sol-Eng+low HFD), were fed a high fat diet for six months. Plasma samples were used for biochemical, ELISA and Luminex analyses of total cholesterol, sEng and inflammatory markers. Functional parameters of aorta were assessed with wire myograph 620M. Western blot analyses of membrane endoglin/eNOS signaling and endothelial dysfunction/inflammation markers in aorta were performed. RESULTS: Functional analysis of aorta showed impaired KCl induced vasoconstriction, endothelial-dependent relaxation after the administration of acetylcholine as well as endothelial-independent relaxation induced by sodium nitroprusside in the Sol-Eng+high HFD group compared to the Sol-Eng+low HFD group. Ach-induced vasodilation after administration of l-NAME was significantly higher in the Sol-Eng+high HFD group compared to the Sol-Eng+low HFD group. The expression of endoglin, p-eNOS/eNOS, pSmad2/3/Smad2/3 signaling pathway was significantly lower in the Sol-Eng+high HFD group compared to the Sol-Eng+low HFD group. CONCLUSIONS: The results indicate that long-term hypercholesterolemia combined with high levels of sEng leads to the aggravation of endothelial and vessel wall dysfunction in aorta, with possible alterations of the membrane endoglin/eNOS signaling, suggesting that high levels of soluble endoglin might be considered as a risk factor of cardiovascular diseases.
- Klíčová slova
- Endothelial dysfunction, Mice, Soluble endoglin, Vascular reactivity, eNOS,
- MeSH
- aorta metabolismus patologie patofyziologie MeSH
- ateroskleróza genetika metabolismus patologie patofyziologie MeSH
- cévní endotel metabolismus patologie patofyziologie MeSH
- dieta s vysokým obsahem tuků MeSH
- endoglin genetika metabolismus MeSH
- fosforylace MeSH
- hypercholesterolemie komplikace etiologie MeSH
- lidé MeSH
- mediátory zánětu metabolismus MeSH
- modely nemocí na zvířatech MeSH
- myši inbrední C57BL MeSH
- myši inbrední CBA MeSH
- myši transgenní MeSH
- nemoci aorty genetika metabolismus patologie patofyziologie MeSH
- protein Smad2 metabolismus MeSH
- protein Smad3 metabolismus MeSH
- signální transdukce MeSH
- synthasa oxidu dusnatého, typ III metabolismus MeSH
- upregulace MeSH
- vazodilatace MeSH
- vazokonstrikce MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- endoglin MeSH
- ENG protein, human MeSH Prohlížeč
- mediátory zánětu MeSH
- Nos3 protein, mouse MeSH Prohlížeč
- protein Smad2 MeSH
- protein Smad3 MeSH
- Smad2 protein, mouse MeSH Prohlížeč
- Smad3 protein, mouse MeSH Prohlížeč
- synthasa oxidu dusnatého, typ III MeSH
A soluble form of endoglin (sEng) released into the circulation was suggested to be a direct inducer of endothelial dysfunction, inflammation and contributed to the development of hypertension by interfering with TGF-β signaling in cardiovascular pathologies. In the present study, we assessed the hypothesis that high sEng level-induced hypertension via a possible sEng interference with TGF-β signaling pathways may result in inflammatory, structural or fibrotic changes in hearts of Sol-Eng+ mice (mice with high levels of soluble endoglin) fed either chow or high-fat diet. Female Sol-Eng+ mice and their age matched littermates with low plasma levels of sEng were fed either chow or high-fat diet (HFD). Heart samples were subsequently analyzed by histology, qRT-PCR and Western blot analysis. In this study, no differences in myocardial morphology/hypertrophy and possible fibrotic changes between Sol-Eng+ mice and control mice were detected on both chow and HFD. The presence of sEng did not significantly affect the expression of selected members of TGF-β signaling (membrane endoglin, TGFβRII, ALK-5, ALK-1, Id-1, PAI-1 and activated Smad proteins-pSmad 1,5 and pSmad 2,3), inflammation, heart remodeling (PDGFb, Col1A1) and endothelial dysfunction (VCAM-1, ICAM-1) in the hearts of Sol-Eng+ mice compared to control mice on both chow and high-fat diet. High levels of soluble endoglin did not affect microscopic structure (profibrotic and degenerative cardiomyocyte changes), and specific parts of TGF-β signaling, endothelial function and inflammation in the heart of Sol-Eng+ mice fed both chow diet or HFD. However, we cannot rule out a possibility that a long-term chronic exposure (9 months and more) to soluble endoglin alone or combined with other cardiovascular risk factors may contribute to alterations of heart function and structure in Sol-Eng+ mice, which is the topic in our lab in ongoing experiments.
- Klíčová slova
- Heart, Inflammation, Mice, Soluble endoglin, TGF-β signaling,
- MeSH
- endoglin biosyntéza genetika MeSH
- hypertenze krev komplikace MeSH
- hypertrofická kardiomyopatie krev etiologie patologie MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- modely nemocí na zvířatech MeSH
- myokard metabolismus patologie MeSH
- myši inbrední C57BL MeSH
- myši transgenní MeSH
- myši MeSH
- oxidační stres MeSH
- regulace genové exprese * MeSH
- RNA genetika MeSH
- signální transdukce MeSH
- srdce MeSH
- stanovení celkové genové exprese MeSH
- transformující růstový faktor beta genetika metabolismus MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- endoglin MeSH
- Eng protein, mouse MeSH Prohlížeč
- RNA MeSH
- transformující růstový faktor beta MeSH
AIMS: A soluble form of endoglin (sEng) was proposed to participate in the induction of endothelial dysfunction in small blood vessels. Here, we tested the hypothesis that high levels of sEng combined with a high-fat diet induce endothelial dysfunction in an atherosclerosis-prone aorta. METHODS AND RESULTS: Six-month-old female and male transgenic mice overexpressing human sEng (Sol-Eng+) with low (Sol-Eng+low) or high (Sol-Eng+high) levels of plasma sEng were fed a high-fat rodent diet containing 1.25% cholesterol and 40% fat for 3 months. The plasma cholesterol and mouse sEng levels did not differ in the Sol-Eng+high and Sol-Eng+low mice. The expression of proinflammatory (P-selectin, ICAM-1, pNFκB and COX-2) and oxidative-stress-related markers (HO-1, NOX-1 and NOX-2) in the aortas of Sol-Eng+high female mice was significantly higher than in Sol-Eng+low female mice. Endothelium-dependent vasodilatation induced by acetylcholine was preserved better in the Sol-Eng+ high female mice than in the Sol-Eng+low female mice. CONCLUSION: These results suggest that high concentrations of sEng in plasma in combination with a high-fat diet induce the simultaneous activation of proinflammatory, pro-oxidative and vasoprotective mechanisms in mice aorta and the balance of these biological processes determines whether the final endothelial phenotype is adaptive or maladaptive.
- MeSH
- aorta účinky léků metabolismus patofyziologie MeSH
- ateroskleróza krev genetika metabolismus patofyziologie MeSH
- biologické markery metabolismus MeSH
- dieta s vysokým obsahem tuků * MeSH
- endoglin krev genetika metabolismus MeSH
- fenotyp MeSH
- fyziologická adaptace MeSH
- genetická predispozice k nemoci MeSH
- lidé MeSH
- mediátory zánětu metabolismus MeSH
- modely nemocí na zvířatech MeSH
- myši inbrední C57BL MeSH
- myši inbrední CBA MeSH
- myši transgenní MeSH
- nemoci aorty krev genetika metabolismus patofyziologie MeSH
- oxid dusnatý metabolismus MeSH
- oxidační stres * MeSH
- upregulace MeSH
- vazodilatace * účinky léků MeSH
- vazodilatancia farmakologie MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zánět krev genetika metabolismus patofyziologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- biologické markery MeSH
- endoglin MeSH
- ENG protein, human MeSH Prohlížeč
- mediátory zánětu MeSH
- oxid dusnatý MeSH
- vazodilatancia MeSH
