32 research outputs found
Additional file 3: Figure S1. of Oxidized LDL-induced JAB1 influences NF-ÎşB independent inflammatory signaling in human macrophages during foam cell formation
OxLDL influence JAB1 protein expression and foam cell formation in differentiated human MÎŚ. (PDF 258Â kb
Additional file 2: of Oxidized LDL-induced JAB1 influences NF-ÎşB independent inflammatory signaling in human macrophages during foam cell formation
Primers used in this study. (PDF 134Â kb
Additional file 5: of Oxidized LDL-induced JAB1 influences NF-κB independent inflammatory signaling in human macrophages during foam cell formation
OxLDL-induced NF-κB signaling in human MФ after 24 h. (PDF 340 kb
Additional file 4: of Oxidized LDL-induced JAB1 influences NF-ÎşB independent inflammatory signaling in human macrophages during foam cell formation
Knock-down of JAB1 in human MФ. (PDF 264 kb
Additional file 1: of Lower hypoxic ventilatory response in smokers compared to non-smokers during abstinence from cigarettes
Supportive information. (XLSX 22Â kb
Image2_PACAP regulates VPAC1 expression, inflammatory processes and lipid homeostasis in M1- and M2-macrophages.tiff
BackgroundPituitary adenylate cyclase-activating polypeptide (PACAP) acts as an anti-atherogenic neuropeptide and plays an important role in cytoprotective, as well as inflammatory processes, and cardiovascular regulation. Therefore, the aim of this study is to investigate the regulatory effects of PACAP and its receptor VPAC1 in relation to inflammatory processes and lipid homeostasis in different macrophage (MΦ) subtypes.MethodsTo investigate the role of PACAP deficiency in the pathogenesis of atherosclerosis under standard chow (SC) or cholesterol-enriched diet (CED) in vivo, PACAP−/− mice were crossbred with ApoE−/− to generate PACAP−/−/ApoE−/− mice. Lumen stenosis in the aortic arch and different MΦ-subtypes were analyzed in atherosclerotic plaques by quantitative immunohistochemistry. Undifferentiated bone marrow-derived cells (BMDC) from 30-weeks-old ApoE−/− and PACAP−/−/ApoE−/− mice were isolated, differentiated into BMDM1- and BMDM2-MΦ, and incubated with oxidized low-density lipoprotein (oxLDL). In addition, PMA-differentiated human THP-1 MΦ were further differentiated into M1-/M2-MΦ and subsequently treated with PACAP38, the VPAC1 agonist [(Ala11,22,28)VIP], the antagonist (PG 97–269), and/or oxLDL. Uptake/accumulation of oxLDL was analyzed by oxLDL-DyLight™488 and Bodipy™ 493/503. The mRNA expression was analyzed by qRT-PCR, protein levels by Western blot, and cytokine release by ELISA.ResultsIn vivo, after 30 weeks of SC, PACAP−/−/ApoE−/− mice showed increased lumen stenosis compared with ApoE−/− mice. In atherosclerotic plaques of PACAP−/−/ApoE−/− mice under CED, immunoreactive areas of VPAC1, CD86, and CD163 were increased compared with ApoE−/− mice. In vitro, VPAC1 protein levels were increased in PACAP−/−/ApoE−/− BMDM compared with ApoE−/− BMDM, resulting in increased TNF-α mRNA expression in BMDM1-MΦ and decreased TNF-α release in BMDM2-MΦ. Concerning lipid homeostasis, PACAP deficiency decreased the area of lipid droplets in BMDM1-/M2-MΦ with concomitant increasing adipose differentiation-related protein level. In THP-1 M1-/M2-MΦ, the VPAC1 antagonist increased the uptake of oxLDL, whereas the VPAC1 agonist decreased the oxLDL-induced intracellular triglyceride content.ConclusionOur data suggest that PACAP via VPAC1 signaling plays an important regulatory role in inflammatory processes in atherosclerotic plaques and in lipid homeostasis in different MΦ-subtypes, thereby affecting foam cell formation. Therefore, VPAC1 agonists or PACAP may represent a new class of anti-atherogenic therapeutics.</p
Image1_PACAP regulates VPAC1 expression, inflammatory processes and lipid homeostasis in M1- and M2-macrophages.tiff
BackgroundPituitary adenylate cyclase-activating polypeptide (PACAP) acts as an anti-atherogenic neuropeptide and plays an important role in cytoprotective, as well as inflammatory processes, and cardiovascular regulation. Therefore, the aim of this study is to investigate the regulatory effects of PACAP and its receptor VPAC1 in relation to inflammatory processes and lipid homeostasis in different macrophage (MΦ) subtypes.MethodsTo investigate the role of PACAP deficiency in the pathogenesis of atherosclerosis under standard chow (SC) or cholesterol-enriched diet (CED) in vivo, PACAP−/− mice were crossbred with ApoE−/− to generate PACAP−/−/ApoE−/− mice. Lumen stenosis in the aortic arch and different MΦ-subtypes were analyzed in atherosclerotic plaques by quantitative immunohistochemistry. Undifferentiated bone marrow-derived cells (BMDC) from 30-weeks-old ApoE−/− and PACAP−/−/ApoE−/− mice were isolated, differentiated into BMDM1- and BMDM2-MΦ, and incubated with oxidized low-density lipoprotein (oxLDL). In addition, PMA-differentiated human THP-1 MΦ were further differentiated into M1-/M2-MΦ and subsequently treated with PACAP38, the VPAC1 agonist [(Ala11,22,28)VIP], the antagonist (PG 97–269), and/or oxLDL. Uptake/accumulation of oxLDL was analyzed by oxLDL-DyLight™488 and Bodipy™ 493/503. The mRNA expression was analyzed by qRT-PCR, protein levels by Western blot, and cytokine release by ELISA.ResultsIn vivo, after 30 weeks of SC, PACAP−/−/ApoE−/− mice showed increased lumen stenosis compared with ApoE−/− mice. In atherosclerotic plaques of PACAP−/−/ApoE−/− mice under CED, immunoreactive areas of VPAC1, CD86, and CD163 were increased compared with ApoE−/− mice. In vitro, VPAC1 protein levels were increased in PACAP−/−/ApoE−/− BMDM compared with ApoE−/− BMDM, resulting in increased TNF-α mRNA expression in BMDM1-MΦ and decreased TNF-α release in BMDM2-MΦ. Concerning lipid homeostasis, PACAP deficiency decreased the area of lipid droplets in BMDM1-/M2-MΦ with concomitant increasing adipose differentiation-related protein level. In THP-1 M1-/M2-MΦ, the VPAC1 antagonist increased the uptake of oxLDL, whereas the VPAC1 agonist decreased the oxLDL-induced intracellular triglyceride content.ConclusionOur data suggest that PACAP via VPAC1 signaling plays an important regulatory role in inflammatory processes in atherosclerotic plaques and in lipid homeostasis in different MΦ-subtypes, thereby affecting foam cell formation. Therefore, VPAC1 agonists or PACAP may represent a new class of anti-atherogenic therapeutics.</p
Value of electric impedance measurements in detecting inflammatory processes defined by different markers of inflammation.
<p>Value of electric impedance measurements in detecting inflammatory processes defined by different markers of inflammation.</p
Cross section depicting an advanced lesion type Va.
<p>Movats staining, magnification 20×. LC: Characteristic lipid core. FC: Thick fibrous cap with yellow staining of collagen fibers and red-colored smooth muscle cells. M: Media. Arrow: Microhemorrhages, typically found at the lateral margin of the lipid core (red: stained fibrin).</p
Experimental setup of the impedance measurement system.
<p>Experimental setup of the impedance measurement system.</p