Effect of carbenoxolone on adipose tissue gene expression quantified by semi quantitative reverse transcription PCR.

<p>(A). Stearoyl CoA desaturase 1 (SCD1). (B). Malic enzyme 1 (ME1). (C). Macrophage expressed gene (MPEG). (D). Lysosomal acid lipase (LIPA). (E). Beta3-Adrenergic receptor (β3-AR). Parameters were measured after 4weeks of treatment with carbenoxolone or vehicle (50 mg/kg body weight/day). Empty bars indicate lean phenotype where as filled bars indicate obese phenotype. Relative gene expression was measured using calnexin gene as internal control. Relative expression of gene of interest in lean control was taken as 1. Values are means ± SEM for 4 animals for group. . #<i>p</i><0.05, ##<i>p</i><0.01 and ### <i>p</i><0.001 comparing vehicle-treated lean and obese rats. *<i>p</i><0.05, **<i>p</i><0.01 and *** <i>p</i><0.001 comparing carbenoxolone-treated animals with vehicle-treated animals of the same phenotype.</p

Effect of carbenoxolone on adipose tissue morphology and glycogen content.

<p>(A). Adipocyte size (number of cells/16sq.mm). (B). Photographs of adipose tissue sections stained with haematoxylin and eosin (to observe adipocyte cell size). (C). Photographs of adipose tissue sections with Haematoxylin and Eosin (to observe adipose tissue inflammation characterized by presence of ‘crown like structures’-indicated by arrows). (D). Photographs of adipose tissue sections stained with Masson's trichrome stain (to observe fibrosis-indicated by arrows). (E). Tissue glycogen content. Parameters were measured after 4weeks of treatment with carbenoxolone or vehicle (50 mg/kg body weight/day). Empty bars indicate lean phenotype, where as filled bars indicate obese phenotype. OC, Obese control; OT, Obese- treated. Images were taken at Magnification of 10X. Values are means ± SEM for 6 animals for group. . #<i>p</i><0.05, ##<i>p</i><0.01 and ### <i>p</i><0.001 comparing vehicle-treated lean and obese rats. *<i>p</i><0.05, **<i>p</i><0.01 and *** <i>p</i><0.001 comparing carbenoxolone-treated animals with vehicle-treated animals of the same phenotype.</p

Effect of carbenoxolone on body composition of WNIN/Ob lean and obese rats.

<p>(A). fat percentage. (B). Lean body mass. (C). Fat free mass. Body composition was measured by TOBEC after 4weeks of treatment with carbenoxolone or vehicle (50 mg/kg body weight/day). Empty bars indicate lean phenotype where as filled bars indicate obese phenotype. Values are means ± SEM for 6 animals for group. #<i>p</i><0.05, ##<i>p</i><0.01 and ### <i>p</i><0.001 comparing vehicle-treated lean and obese rats. *<i>p</i><0.05, **<i>p</i><0.01 and *** <i>p</i><0.001 comparing carbenoxolone-treated animals with vehicle-treated animals of the same phenotype.</p

Effect of carbenoxolone on food intake, body weight and organ weights in WNIN/Ob lean and obese rats.

<p>Values represent means ± SEM of 6 rats per group. LC, Lean control; OC, Obese control; LT, Lean- treated; OT, Obese- treated. Parameters were measured after four weeks of treatment with carbenoxolone (50 mg/kg/body weight/day) or vehicle (same volume of phosphate buffer saline).</p>#<p><i>p</i><0.05,</p>##<p><i>p</i><0.01 and ### <i>p</i><0.001 comparing vehicle-treated lean and obese rats.</p>*<p><i>p</i><0.05,</p>**<p><i>p</i><0.01 and *** <i>p</i><0.001 comparing carbenoxolone- treated animals with vehicle-treated animals of the same phenotype. </p

Effect of carbenoxolone on insulin signaling in gastrocnemius muscle of WNIN/Ob lean and obese rats.

<p>(A). Representative western blot for AKT. (B). Representative western blot for pAKT (Ser 473). (C). Representative western blot for PTP1B. (C). Representative western blot for β-actin. Parameters were measured after 4weeks of treatment with carbenoxolone or vehicle (50 mg/kg body weight/day). Relative protein levels were measured using β-actin as internal control. LC, Lean Control; LT, Lean Treated; OC, Obese Control; OT, Obese control. Relative expression of protein of interest in lean control was taken as 1. Values are means ± SEM for 4 animals for group. . #<i>p</i><0.05 comparing vehicle-treated lean and obese rats. *<i>p</i><0.05 comparing carbenoxolone-treated animals with vehicle-treated animals of the same phenotype.</p

Effect of carbenoxolone on plasma biochemical parameters in WNIN/Ob lean and obese rats.

<p>(A). Fed-state corticosterone. (B). Fasting corticosterone. (C). Fed-state triglycerides. (D). Fasting triglycerides. (E). Fed-state total cholesterol. (F). Fasting total cholesterol. (G). Fed-state HDL cholesterol. (H). Fasting HDL cholesterol. Plasma parameters were measured after 4weeks of treatment with carbenoxolone or vehicle (50 mg/kg bodyweight/day). Empty bars indicate lean phenotype where as filled bars indicate obese phenotype. Values are means ± SEM for 6 animals for group. #<i>p</i><0.05, ##<i>p</i><0.01 and ### <i>p</i><0.001 comparing vehicle-treated lean and obese rats. *<i>p</i><0.05, **<i>p</i><0.01 and *** <i>p</i><0.001 comparing carbenoxolone-treated animals with vehicle-treated animals of the same phenotype.</p

Effect of carbenoxolone on time course of plasma insulin (A) and glucose (B) during oral glucose tolerance test.

<p>Parameters were measured after 4weeks of treatment with carbenoxolone or vehicle (50 mg/kg body weight/day). Values are means ± SEM for 6 animals for group. #<i>p</i><0.05, ##<i>p</i><0.01 and ### <i>p</i><0.001 comparing vehicle-treated lean and obese rats at the respective time point. *<i>p</i><0.05, **<i>p</i><0.01 and *** <i>p</i><0.001 comparing carbenoxolone-treated animals with vehicle-treated animals of the same phenotype at the respective time point.</p

Effect of carbenoxolone on hepatic triglyceride and glycogen content.

<p>(A). Hepatic triglyceride content. (B). Hepatic glycogen content. (C). Photographs of liver tissue sections stained with haematoxylin and eosin (to study fat accumulation). (D). Photographs of liver tissue sections stained with oil Red O (to study fat accumulation). (E). Photographs of liver tissue sections stained with periodic acid-Schiff's stain (to study glycogen content). (F). Photographs of liver tissues sections stained with Masson's trichrome stain (to observe fibrosis). Parameters were measured after 4weeks of treatment with carbenoxolone or vehicle (50 mg/kg body weight/day). Empty bars indicate lean phenotype where as filled bars indicate obese phenotype. LC, Lean control; LT, Lean- treated; OC, Obese control; OT, Obese -treated. Images were taken at Magnification of 10X. Values are means ± SEM for 6 animals for group. #<i>p</i><0.05, ##<i>p</i><0.01 and ### <i>p</i><0.001 comparing vehicle-treated lean and obese rats. *<i>p</i><0.05, **<i>p</i><0.01 and *** <i>p</i><0.001 comparing carbenoxolone-treated animals with vehicle-treated animals of the same phenotype.</p

Effect of carbenoxolone on 11β-HSD1 activity in WNIN/Ob lean and obese rats.

<p>(A). Liver. (B). Subcutaneous adipose tissue. (C). Omental adipose tissue. (D). Skeletal muscle. 11β-HSD1 activity was measured after 1 h of subcutaneous injection of carbenoxolone or vehicle (50 mg/kg body weight/day). Empty bars indicate lean phenotype where as filled bars indicate obese phenotype. Values are means ± SEM for 6 animals for group. #<i>p</i><0.05, ##<i>p</i><0.01 and ### <i>p</i><0.001 comparing vehicle-treated lean and obese rats. *<i>p</i><0.05, **<i>p</i><0.01 and *** <i>p</i><0.001 comparing carbenoxolone-treated animals with vehicle-treated animals of the same phenotype.</p