Journal: bioRxiv
Article Title: Gut Microbiota Production of Phenylacetate Programs Vascular Niche Senescence and Drives Atherosclerosis
doi: 10.64898/2026.02.27.708541
Figure Lengend Snippet: a,b, Plasma samples from aged ASCVD patients (>80 years old) enrolled in the ASCVD cohort ( n = 110; male and female) and healthy controls ( n = 77; male and female) were subjected to targeted metabolomics for PAA quantification. c, Adjusted regression models for the association of PAA with atherosclerosis in the ASCVD cohort ( n □=□187). Effect estimates were controlled for age, sex, smoking, alcohol, family history of CVD, LDL-C, triglycerides, HDL-C, Hb1Ac, CRP, troponin T, and NT-proBNP. Error bars show 95% confidence intervals. OR, odds ratio. d, Ldlr −/− and WT mice were fed chow (for 8 weeks) or Western diet (for 12 weeks). WC: WT + chow diet; WW: WT + Western diet; LC: Ldlr −/− + chow diet; LW: Ldlr −/− + Western diet. e, Plasma PAA levels in these mice quantified by LC-MS/MS targeted metabolomics ( n □=□5). f, Representative images (left) and quantification (right) of H&E staining of aortic root lesions ( n □=□5). Arrowheads indicate plaque areas. g, Correlation of plasma PAA levels with aortic root lesion area and luminal occlusion (%). h, Representative immunoblots and quantification of intensities for the CDK inhibitor p16 INK4A , the SASP component IL-6, DNA damage marker γ-H2A.X, and the endothelial function marker phosphorylated eNOS S1177 in aortas from Ldlr −/− and WT mice fed a chow or Western diet for 12 weeks ( n = 5). i, Expression of CDKN1A , IL1B , and IL6 genes (upper) and co-expression of the senescence-associated genes in atherosclerotic aortic wall (AOR) of patients with CAD ( n □=□600) and healthy individuals ( n □=□250) in the STARNET database. j,k,l, Representative immunoblots and quantification of intensities for senescence hallmarks ( j ), insulin signaling pathway ( k ), and thermogenic markers ( l ) in tPVAT from Ldlr −/− and WT mice fed a chow or Western diet for 12 weeks ( n = 5). m, Plasma and aortic PVAT samples were collected from the validation study, including aged CAD patients undergoing CABG surgery ( n □=□5) and non-CAD controls ( n □=□5), for LC-MS/MS PAA quantification and senescence studies. n, Plasma PAA concentrations in aged CAD patients ( n □=□5) and non-CAD controls ( n □=□5). o,p,q, Representative immunoblots and quantification of intensities for senescence hallmarks ( o ), NOTCH1 and thermogenic markers ( p ), and insulin signaling pathway ( q ) in aortic PVAT from the individuals in the validation study ( n = 5). r, PAA was administered (PAA) or not (Ctrl) to chow-fed Ldlr −/− mice for 8 weeks. s, Quantification of H&E-stained aortic root lesion area (left) and aortic occlusion (right). Total cholesterol concentrations in plasma ( n = 5). t, Representative immunoblots and quantification of intensities for senescence hallmarks p16 INK4A , IL-6, and γ-H2A.X in aortas (left) and tPVAT (right). Data were determined in 8-10 micrographs and represent triplicated biologically independent experiments. Scale bar, 200 μm ( f ). Error bars represent SD ( h,j-l,o-t ). P values were calculated using two-tailed Mann–Whitney U -test ( b ), one-way ANOVA followed by Tukey’s post hoc test ( e,f,h,j-l ), Welch’s t-test ( i ), and a two-tailed unpaired Student’s t -test ( n-t ). Correlation coefficient and P values were calculated by Spearman’s rank-order correlation test ( g ). Data are shown as median with min–max; each violin represents interquartile range (IQR); center lines indicate the median; upper and lower lines are bounded by 25th and 75th percentiles ( b , n ). Images created with https://BioRender.com ( a,d,m,r ).
Article Snippet: Cell culture supernatants were assessed for IL-6 using a QuantikineTM ELISA Human IL-6 Immunoassay kit (R&D Systems, D6050), according to the manufacturer’s instructions.
Techniques: Clinical Proteomics, Western Blot, Liquid Chromatography with Mass Spectroscopy, Staining, Marker, Expressing, Biomarker Discovery, Two Tailed Test, MANN-WHITNEY
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