Figure 1 Fructose, fatty acids, and ectopic fat. VAT, visceral adipose tissue. Figure 1. Fructose, fatty acids, and ectopic fat. VAT, visceral adipose tissue. More

Figure 3 Mean SUV for glucose of various tissues during CE and TN conditions. SQAT, subcutaneous adipose tissue; VAT, visceral adipose tissue; white bars, TN; black bars, CE. Data are mean ± SEM. *P < 0.05, **P < 0.01, CE vs. TN. Figure 3. Mean SUV for glucose of various tissues during CE and TN conditions. SQAT, subcutaneous adipose tissue; VAT, visceral adipose tissue; white bars, TN; black bars, CE. Data are mean ± SEM. *P < 0.05, **P < 0.01, CE vs. TN. More

FIG. 3. The change in fat mass (A), the change in glucose uptake in adipose tissue per tissue weight (B), and the change in glucose uptake in whole-fat depot (C). Vertical bars denote SD. □, placebo; ▪, metformin; [cjs2108], rosiglitazone. FemAT, femoral subcutaneous adipose tissue; SCAAT, subcutaneous abdominal adipose tissue; VAT, visceral adipose tissue. §P < 0.05 for the change vs. placebo. Asterisks (*) shown within the columns are for the changes (P < 0.05) within the group. FIG. 3. The change in fat mass (A), the change in glucose uptake in adipose tissue per tissue weight (B), and the change in glucose uptake in whole-fat depot (C). Vertical bars denote SD. □, placebo; ▪, metformin; [cjs2108], rosiglitazone. FemAT, femoral subcutaneous adipose tissue; SCAAT, subcutaneous abdominal adipose tissue; VAT, visceral adipose tissue. §P < 0.05 for the change vs. placebo. Asterisks (*) shown within the columns are for the changes (P < 0.05) within the group. More

FIG. 1. Relationships of the SNP rs738409 C>G (Ile148Met) in PNPLA3 with liver fat (A) and insulin sensitivity (B) in all 330 subjects (TAT, total adipose tissue; VAT, visceral adipose tissue). Liver fat content in subjects without and with fatty liver and the relationship of the SNP rs738409 with liver fat content in subjects having fatty liver (C). Insulin sensitivity in subjects without and with fatty liver and the relationship of the SNP rs738409 with insulin sensitivity in subjects having fatty liver (D). FIG. 1. Relationships of the SNP rs738409 C>G (Ile148Met) in PNPLA3 with liver fat (A) and insulin sensitivity (B) in all 330 subjects (TAT, total adipose tissue; VAT, visceral adipose tissue). Liver fat content in subjects without and with fatty liver and the relationship of the SNP rs738409 with liver fat content in subjects having fatty liver (C). Insulin sensitivity in subjects without and with fatty liver and the relationship of the SNP rs738409 with insulin sensitivity in subjects having fatty liver (D). More

Figure 7 Representative Western blot analysis of ITCH protein in adipose tissue (AT), isolated adipocytes (adipocyte fraction [AF]), and SVFs (A). Representative Western blot analysis of ITCH protein in AT, isolated adipocytes (AF), and SVFs (B). Images are representative of adipose tissue sections collected from five subjects. CLS, crown-like structure; ma, macrophage. The arrows indicate ITCH- and CD3-positive cells. Correlation between Itch and CD-206 mRNA (C) expression (r = −0.36, P = 0.01) and CD206-to-CD68 ratio (D) (r = −0.42, P = 0.003) in human adipose tissue (n = 50). Expression of mRNA was determined by real-time PCR and normalized to cyclophilin A. R.U., relative units; VAT, visceral adipose tissue. Figure 7. Representative Western blot analysis of ITCH protein in adipose tissue (AT), isolated adipocytes (adipocyte fraction [AF]), and SVFs (A). Representative Western blot analysis of ITCH protein in AT, isolated adipocytes (AF), and SVFs (B). Images are representative of adipose tissue sections collected from five subjects. CLS, crown-like structure; ma, macrophage. The arrows indicate ITCH- and CD3-positive cells. Correlation between Itch and CD-206 mRNA (C) expression (r = −0.36, P = 0.01) and CD206-to-CD68 ratio (D) (r = −0.42, P = 0.003) in human adipose tissue (n = 50). Expression of mRNA was determined by real-time PCR and normalized to cyclophilin A. R.U., relative units; VAT, visceral adipose tissue. More

Figure 2 TREM2 deficiency aggravates obesity-induced insulin resistance. A: Insulin tolerance test (ITT) of WT and Trem2^–/– mice 13 weeks post-HFD (n = 8 per genotype). B: Area under the curve (AUC) of panel A. C: Oral glucose tolerance test (oGTT) of WT and Trem2^–/– mice 13 weeks post-HFD (n = 8 per genotype). D: Adipose weights 13 weeks post-HFD of animals in panel C. E: Mouse weights 13 weeks post-HFD of animals in panels C and D. F–H: Energy expenditure and activity of control and Trem2^–/– mice 13 weeks post-HFD (n = 4 per genotype). Data are mean ± SEM and are pooled for panels A–E from two independent experiments. Statistical analysis was performed with two-way ANOVA followed by Bonferroni posttest (A and C), Student t test (B and D), or one-way ANOVA followed by Tukey posttest (G). *P < 0.05, **P < 0.01. AU, arbitrary unit; RP, retroperitoneal; SAT, subcutaneous adipose tissue; VAT, visceral adipose tissue. Figure 2. TREM2 deficiency aggravates obesity-induced insulin resistance. A: Insulin tolerance test (ITT) of WT and Trem2–/– mice 13 weeks post-HFD (n = 8 per genotype). B: Area under the curve (AUC) of panel A. C: Oral glucose tolerance test (oGTT) of WT and Trem2–/– mice 13 weeks post-HFD (n = 8 per genotype). D: Adipose weights 13 weeks post-HFD of animals in panel C. E: Mouse weights 13 weeks post-HFD of animals in panels C and D. F–H: Energy expenditure and activity of control and Trem2–/– mice 13 weeks post-HFD (n = 4 per genotype). Data are mean ± SEM and are pooled for panels A–E from two independent experiments. Statistical analysis was performed with two-way ANOVA followed by Bonferroni posttest (A and C), Student t test (B and D), or one-way ANOVA followed by Tukey posttest (G). *P < 0.05, **P < 0.01. AU, arbitrary unit; RP, retroperitoneal; SAT, subcutaneous adipose tissue; VAT, visceral adipose tissue. More

FIG. 6. Smad3-KO mice are resistant to HFD-induced obesity and hepatic steatosis. A and B: Graph shows mean body weight of WT and KO mice on HFD or LFD over 18 weeks (A) and at the end of the 18-week diet regimen (B). C: Representative images of subcutaneous, epididymal, and visceral adipose tissues, and the liver of WT and KO mice from the two diet regimens. Small division on the ruler scale = 1 mm. D: Mean weights of the liver and the indicated adipose tissues of WT and KO mice after 18 weeks on HFD or LFD. Values are expressed as percentage of body weight. E: Representative hematoxylin–eosin-stained histologic section of the WT and KO EAT after 18 weeks on HFD or LFD treatment. Scale bars, 100 μm. F: Mean cross-sectional area of adipocytes (n= 400) from the WT and KO mice on the indicated diet regimen. G: Representative histologic section (oil red O and methylene blue staining) of the liver tissues. Scale bars, 100 μm. White arrows indicate oil droplets. H–J: Quantification of lipid accumulation in peripheral organs. Total triglyceride extracted from fresh-frozen liver (H), skeletal muscle (I), and pancreas (J) of WT and KO mice fed on HFD or LFD diet was expressed as amount in milimole per 1 g of protein. Data represent mean ± SEM, n = 10/group. ##P < 0.01 vs. WT LFD control; *P < 0.05, **P < 0.01, ***P < 0.001. EAT, epididymal adipose tissue; SAT, subcutaneous adipose tissue; VAT, visceral adipose tissue. (A high-quality digital representation of this figure is available in the online issue.) FIG. 6. Smad3-KO mice are resistant to HFD-induced obesity and hepatic steatosis. A and B: Graph shows mean body weight of WT and KO mice on HFD or LFD over 18 weeks (A) and at the end of the 18-week diet regimen (B). C: Representative images of subcutaneous, epididymal, and visceral adipose tissues, and the liver of WT and KO mice from the two diet regimens. Small division on the ruler scale = 1 mm. D: Mean weights of the liver and the indicated adipose tissues of WT and KO mice after 18 weeks on HFD or LFD. Values are expressed as percentage of body weight. E: Representative hematoxylin–eosin-stained histologic section of the WT and KO EAT after 18 weeks on HFD or LFD treatment. Scale bars, 100 μm. F: Mean cross-sectional area of adipocytes (n= 400) from the WT and KO mice on the indicated diet regimen. G: Representative histologic section (oil red O and methylene blue staining) of the liver tissues. Scale bars, 100 μm. White arrows indicate oil droplets. H–J: Quantification of lipid accumulation in peripheral organs. Total triglyceride extracted from fresh-frozen liver (H), skeletal muscle (I), and pancreas (J) of WT and KO mice fed on HFD or LFD diet was expressed as amount in milimole per 1 g of protein. Data represent mean ± SEM, n = 10/group. ##P < 0.01 vs. WT LFD control; *P < 0.05, **P < 0.01, ***P < 0.001. EAT, epididymal adipose tissue; SAT, subcutaneous adipose tissue; VAT, visceral adipose tissue. (A high-quality digital representation of this figure is available in the online issue.) More

Figure 1 Transcriptomic signature of adipose tissue CD14⁺ cells in OB/D patients. A: Genome-wide mRNA expression analysis using microarrays was performed in visceral adipose tissue CD14⁺ cells isolated from OB (n = 6) and OB/D (n = 6) subjects included in group 1. Age and BMI did not differ between the groups. Heat map representation of the genes significantly upregulated (red) or downregulated (green) in adipose tissue CD14⁺ cells isolated from OB and OB/D subjects. Genes are ordered by gene ontology annotations. B and C: Quantitative PCR analyses of pro-IL-1β and NLRP3 in CD14⁺ cells isolated from adipose tissue of NO (n = 4), OB (n = 16), and OB/D (n = 12) subjects. *P < 0.05 versus NO; δP < 0.05 versus OB. D: IL-1β production by CD14⁺ cells isolated from adipose tissue of obese subjects (n = 5) and treated with incremental doses of NLRP3-inhibitor glyburide. *P < 0.05 versus vehicle using Wilcoxon matched pairs test. E: Quantitative PCR analyses of pro-IL-1β and NLRP3 expression in cell fractions prepared from adipose tissue biopsies of obese subjects (n = 10). F: IL-1β secretion by adipose tissue CD14⁺ cells of NO (n = 7), OB (n = 20), and OB/D (n = 15) subjects. Means are shown as horizontal line. δP < 0.05 versus OB. G: Correlations between IL-1β production by adipose tissue CD14⁺ cells isolated from obese subjects (n = 22) and HbA_1c (percentage). The r correlation coefficient and P values obtained by Pearson’s test are indicated. H: Production of IL-1β by adipose tissue explants obtained from obese subjects at the time of RYGP surgery (month 0) and at months 3, 6 (n = 20), and 12 (n = 9). *P < 0.05 versus month 0. I: Comparison of IL-1β production by subcutaneous and visceral adipose tissue explants obtained from the same obese subjects (n = 10). *P < 0.05 using Wilcoxon matched pairs test. All data are shown as mean ± SEM. Ad, adipocytes; ECM, extracellular matrix; Neg, CD14⁻CD3⁻ cells; SAT, subcutaneous adipose tissue; SVF, stroma vascular fraction; Vehi, vehicle; VAT, visceral adipose tissue. Figure 1. Transcriptomic signature of adipose tissue CD14+ cells in OB/D patients. A: Genome-wide mRNA expression analysis using microarrays was performed in visceral adipose tissue CD14+ cells isolated from OB (n = 6) and OB/D (n = 6) subjects included in group 1. Age and BMI did not differ between the groups. Heat map representation of the genes significantly upregulated (red) or downregulated (green) in adipose tissue CD14+ cells isolated from OB and OB/D subjects. Genes are ordered by gene ontology annotations. B and C: Quantitative PCR analyses of pro-IL-1β and NLRP3 in CD14+ cells isolated from adipose tissue of NO (n = 4), OB (n = 16), and OB/D (n = 12) subjects. *P < 0.05 versus NO; δP < 0.05 versus OB. D: IL-1β production by CD14+ cells isolated from adipose tissue of obese subjects (n = 5) and treated with incremental doses of NLRP3-inhibitor glyburide. *P < 0.05 versus vehicle using Wilcoxon matched pairs test. E: Quantitative PCR analyses of pro-IL-1β and NLRP3 expression in cell fractions prepared from adipose tissue biopsies of obese subjects (n = 10). F: IL-1β secretion by adipose tissue CD14+ cells of NO (n = 7), OB (n = 20), and OB/D (n = 15) subjects. Means are shown as horizontal line. δP < 0.05 versus OB. G: Correlations between IL-1β production by adipose tissue CD14+ cells isolated from obese subjects (n = 22) and HbA1c (percentage). The r correlation coefficient and P values obtained by Pearson’s test are indicated. H: Production of IL-1β by adipose tissue explants obtained from obese subjects at the time of RYGP surgery (month 0) and at months 3, 6 (n = 20), and 12 (n = 9). *P < 0.05 versus month 0. I: Comparison of IL-1β production by subcutaneous and visceral adipose tissue explants obtained from the same obese subjects (n = 10). *P < 0.05 using Wilcoxon matched pairs test. All data are shown as mean ± SEM. Ad, adipocytes; ECM, extracellular matrix; Neg, CD14−CD3− cells; SAT, subcutaneous adipose tissue; SVF, stroma vascular fraction; Vehi, vehicle; VAT, visceral adipose tissue. More

Figure 4 IL-22 increases IL-1β release by adipose tissue macrophages in OB/D patients. A: Protein levels of IL-17RA and IL-22RA1 in CD14⁺ cells isolated from peripheral blood mononuclear cells (blood) or visceral adipose tissue of obese subjects and representative flow cytometry histogram of IL-17RA and IL-22RA1 expression on adipose tissue CD14⁺ cells from one OB/D subject. B: IL-1β production by adipose tissue CD14⁺ cells cocultured with autologous adipose tissue CD3⁺ cells upon anti-CD3/anti-CD28 activation (n = 13). *P < 0.05 versus CD14⁺ cells alone using Wilcoxon matched pairs test. C: IL-1β production by adipose tissue CD14⁺ cells cocultured with autologous adipose tissue CD4⁺ cells upon anti-CD3/anti-CD28 activation in NO (n = 3) and OB/D (n = 5) subjects. *P < 0.05 versus CD14⁺ cells alone using Wilcoxon matched pairs test; δP < 0.05 versus NO using Mann-Whitney test. D: IL-1β secretion from adipose tissue CD14⁺ cells stimulated with recombinant human IL-17 and IL-22 alone or combined or IFN-γ for 24 h (n = 6). *P < 0.05 versus control using Wilcoxon matched pairs test. E: IL-1β production by adipose tissue CD14⁺ cells cocultured with autologous CD4⁺ cells upon anti-CD3/anti-CD28 stimulation and in presence of anti-IFN-γ (n = 5), anti-IL-17 (n = 9), or anti-IL-22 (n = 7) neutralizing antibodies or isotype controls. Data are shown as mean ± SEM. *P < 0.05 versus isotype control using Wilcoxon matched pairs test. F: Pro-IL-1β gene expression in MDM treated for 24 h with rIL-22 or T cell–conditioned media in presence of isotype control or anti-IL-22 neutralizing antibody as indicated (n = 5). *P < 0.05 versus isotype using Mann-Whitney test. All data are shown as mean ± SEM. CM, conditioned media; VAT, visceral adipose tissue. Figure 4. IL-22 increases IL-1β release by adipose tissue macrophages in OB/D patients. A: Protein levels of IL-17RA and IL-22RA1 in CD14+ cells isolated from peripheral blood mononuclear cells (blood) or visceral adipose tissue of obese subjects and representative flow cytometry histogram of IL-17RA and IL-22RA1 expression on adipose tissue CD14+ cells from one OB/D subject. B: IL-1β production by adipose tissue CD14+ cells cocultured with autologous adipose tissue CD3+ cells upon anti-CD3/anti-CD28 activation (n = 13). *P < 0.05 versus CD14+ cells alone using Wilcoxon matched pairs test. C: IL-1β production by adipose tissue CD14+ cells cocultured with autologous adipose tissue CD4+ cells upon anti-CD3/anti-CD28 activation in NO (n = 3) and OB/D (n = 5) subjects. *P < 0.05 versus CD14+ cells alone using Wilcoxon matched pairs test; δP < 0.05 versus NO using Mann-Whitney test. D: IL-1β secretion from adipose tissue CD14+ cells stimulated with recombinant human IL-17 and IL-22 alone or combined or IFN-γ for 24 h (n = 6). *P < 0.05 versus control using Wilcoxon matched pairs test. E: IL-1β production by adipose tissue CD14+ cells cocultured with autologous CD4+ cells upon anti-CD3/anti-CD28 stimulation and in presence of anti-IFN-γ (n = 5), anti-IL-17 (n = 9), or anti-IL-22 (n = 7) neutralizing antibodies or isotype controls. Data are shown as mean ± SEM. *P < 0.05 versus isotype control using Wilcoxon matched pairs test. F: Pro-IL-1β gene expression in MDM treated for 24 h with rIL-22 or T cell–conditioned media in presence of isotype control or anti-IL-22 neutralizing antibody as indicated (n = 5). *P < 0.05 versus isotype using Mann-Whitney test. All data are shown as mean ± SEM. CM, conditioned media; VAT, visceral adipose tissue. More

Figure 3 PM reduces obesity-related adiposity and partially prevents the formation of α-dicarbonyls in adipose tissue. A: Representative pictures of histochemical H-E staining of VAT, used for the quantification of cell size (B) and measurement of the diameter of individual adipocytes (C). D: Weight of visceral adipose mass. MGO (E), GO (F), and (G) 3-DG levels in VAT. H: Activity of the GLO1 enzyme in VAT. Data represent the mean ± SEM. N = 11–15 mice per group. Figure 3. PM reduces obesity-related adiposity and partially prevents the formation of α-dicarbonyls in adipose tissue. A: Representative pictures of histochemical H-E staining of VAT, used for the quantification of cell size (B) and measurement of the diameter of individual adipocytes (C). D: Weight of visceral adipose mass. MGO (E), GO (F), and (G) 3-DG levels in VAT. H: Activity of the GLO1 enzyme in VAT. Data represent the mean ± SEM. N = 11–15 mice per group. More

FIG. 4. Correlations between regional adipose tissue glucose uptake rates (rGU) per tissue weight and whole-body glucose disposal rates. Scatter plots for subcutaneous abdominal adipose tissue (SCAAT) (A), visceral adipose tissue (VAT) (B), and femoral subcutaneous adipose tissue (FemAT) (C). PET1, first PET study at baseline; PET2, second PET study after 26 weeks of treatment; RSG, rosiglitazone group. FIG. 4. Correlations between regional adipose tissue glucose uptake rates (rGU) per tissue weight and whole-body glucose disposal rates. Scatter plots for subcutaneous abdominal adipose tissue (SCAAT) (A), visceral adipose tissue (VAT) (B), and femoral subcutaneous adipose tissue (FemAT) (C). PET1, first PET study at baseline; PET2, second PET study after 26 weeks of treatment; RSG, rosiglitazone group. More

FIG. 7. Correlation of adipose tissue angiogenesis with patient phenotype. Angiogenic response to adipose tissue samples was assessed as the percentage of covered SAT (•) or VAT (○) samples. Correlation between the angiogenic response and waist-to-hip ratio (A), average visceral adipocyte surface (B), serum insulin levels (C), and HOMA index (D) was assessed by regression analysis in the 22 nondiabetic patients. AU, arbitrary units. FIG. 7. Correlation of adipose tissue angiogenesis with patient phenotype. Angiogenic response to adipose tissue samples was assessed as the percentage of covered SAT (•) or VAT (○) samples. Correlation between the angiogenic response and waist-to-hip ratio (A), average visceral adipocyte surface (B), serum insulin levels (C), and HOMA index (D) was assessed by regression analysis in the 22 nondiabetic patients. AU, arbitrary units. More