Additional file 1 of Evaluation of the effect of tofogliflozin on the tissue characteristics of the carotid wall—a sub-analysis of the UTOPIA trial

Additional file 1: Tables S1. Between-group comparison of changes in clinical parameters during the treatment period. Table S2. Changes in concomitantly used anti-diabetic agents. Table S3. Changes in concomitantly used cardiovascular medications. Table S4. The changes of GSM-CCA on the basis of tertiles of changes in mean-IMT during observation perio

Effect of LPS on endothelial adhesion molecules in aortic intima.

<p>Aortic intima of wild-type (WT) or adiponectin knockout (Adipo-KO) mice were removed at 4 hours after LPS administration and were subjected to RT-PCR (A) and western blotting (B). Values are mean ± SEM; n = 6 for each group. * <i>P</i><0.05, compared with the values of WT mice with saline treatment; ‡ <i>P</i><0.05, compared with the values of WT mice with LPS administration group.</p

Localization of adiponectin in aorta.

<p>A, Dual-immunofluorescence was performed in wild-type (WT) and adiponectin knockout (Adipo-KO) mice as described in Materials and Methods. B, High magnification images of aortic intima using confocal laser microscope were obtained from WT mice. Green, adiponectin; blue, DAPI; red, CD31.</p

Adiponectin Protein Exists in Aortic Endothelial Cells

<div><p>Aims</p><p>Inflammation is closely associated with the development of atherosclerosis and metabolic syndrome. Adiponectin, an adipose-derived secretory protein, possesses an anti-atherosclerotic property. The present study was undertaken to elucidate the presence and significance of adiponectin in vasculature.</p><p>Methods and Results</p><p>Immunofluorescence staining was performed in aorta of wild-type (WT) mice and demonstrated that adiponectin was co-stained with CD31. Thoracic aorta was cut through and then aortic intima was carefully shaved from aorta. Western blotting showed the existence of adiponectin protein in aortic intima, while there was no adiponectin mRNA expression. Adiponectin knockout (Adipo-KO) and WT mice were administered with a low-dose and short-term lipopolysaccharide (LPS) (1 mg/kg of LPS for 4 hours). The endothelium vascular adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) were highly increased in Adipo-KO mice compared to WT mice after LPS administration.</p><p>Conclusions</p><p>Adiponectin protein exists in aortic endothelium under steady state and may protect vasculature from the initiation of atherosclerosis.</p></div

Existence of adiponectin protein in aortic intima.

<p>Mice were perfused with cold saline to eliminate the contamination of circulating adiponectin. After removing perivascular fat, thoracic aorta was cut through and then intima was carefully shaved from the aorta. A, Western blotting with antibodies against adiponectin, perilipin A, and CD31 in aortic intima. B, Multimeric complexes of adiponectin protein in aortic intima. C, The mRNA level of adiponectin in fat tissue and aortic intima. WT, wild-type mice; KO, adiponectin knockout mice; N.D., not detected.</p

Genes related to circadian rhythm.

<p>PER1 : period homolog 1, ERBB3 : v-erb-b2 erythroblastic leukemia viral oncogene homolog 3, CLOCK : clock homolog, PROK2 : prokineticin 2, CRY2 : cryptochrome 2, CYP7B1 : cytochrome P450, family 7, subfamily B, polypeptide 1, ANNAT : arylalkylamine N-acetyltransferase, CRY1 : cryptochrome 1, PRF1 : perforin 1, HEBP1 : heme binding protein 1, PHPP1 : PH domain and leucine rich repeat protein phosphatase 1, KCNMA1 : potassium large conductance calcium-activated channel, subfamily M, alpha member 1, TIMELESS : timeless homolog, PER2 : Period homolog 2, ATOH7 : atonal homolog 7, ARNTL : aryl hydrocarbon receptor nuclear translocator-like, MAT2A : methionine adenosyltransferase II, alpha, NR1D1 : nuclear receptor subfamily 1, group D, member 1, JUN : jun oncogene, HTR7 : 5-hydroxytryptamine receptor 7, PROKR2 : prokineticin receptor 2.</p

Inflammatory response in Ephrin-B1-knockdown 3T3-L1 adipocytes.

<p>The small inhibitory RNA (siRNA) for Ephrin-B1 was introduced in 3T3-L1 adipocytes as described in the Materials and Methods section. A and B, Ephrin-B1 mRNA (A) and protein (B) levels at 24 hrs (left) and 48 hrs (right) after transfection of control-siRNA or Ephrin-B1-siRNA. C, Changes in adipocytokine mRNA levels by Ephrin-B1-siRNA. 3T3-L1 adipocytes were incubated with or without 1 ng/mL of tumor necrosis factor-α (TNF-α) for 24 hrs after siRNA transfection. <i>EfnB1</i> and EFNB1, Ephrin-B1; TNF-α, tumor necrosis factor-α; <i>Mcp-1</i>, monocyte chemoattractant protein-1; <i>Il-6</i>, interleukin-6. Values are mean±SD; n=3 for each group. *<i>P</i><0.05; **<i>P</i><0.01; ***<i>P</i><0.001. #<i>P</i><0.05; # # #<i>P</i><0.001, compared to TNF-α (-).</p

Core clock genes mRNA levels in obese adipose tissues.

<p>A, Tissue distribution of Per1 mRNA. The Per1 mRNA level of the eye was set at 1, and the mRNA levels of Per1 in other tissues are presented relative to the Per1 mRNA level of the eye. B to J, Changes in adipose core clock genes and adipose-related genes in C57BL/6N (B6) and <i>ob/ob</i> (<i>ob</i>) mice at 8, 25 and 35 weeks of age. n = 3–8 per group. Values are mean ± SEM. **P<0.01; ***P<0.001. ^$P<0.05; ^$$P<0.01; ^$$$P<0.001, compared to 8-week-old B6 mice. ^##P<0.01; ^###P<0.001, compared to 8-week-old <i>ob</i> mice.</p

MOESM2 of Pioglitazone strengthen therapeutic effect of adipose-derived regenerative cells against ischemic cardiomyopathy through enhanced expression of adiponectin and modulation of macrophage phenotype

Additional file 2. ADRCs graft making video

A schematic diagram illustrating the possible role of Ephrin-B1 in the development of adipose inflammation in obesity.

<p>In obese adipose tissues, TNF-α and macrophages repress Ephrin-B1 expression in adipocytes. Suppression of Ephrin-B1 in adipocytes augments MCP-1 expression and accelerates monocytes recruitment into adipose tissues. Ephrin-B1 may play an important role in adipose vicious cycle in obesity; reduction of adipose Ephrin-B1 expression in obesity could accelerate the vicious cycle involved in adipose tissue inflammation.</p