Intracellular Ca²⁺ and intracellular Ca²⁺ regulatory protein properties in hearts from low fat (LF) or high fat (HF)-fed C57 mice with or without TUDCA treatment (300 mg/kg for 15 days).

<p>A: Representative traces depicting intracellular Ca²⁺ transients in LF and HF-fed mice; B: Representative traces depicting Ca²⁺ transients in LF and HF-fed mice with TUDCA treatment; C: Baseline intracellular Ca²⁺ levels; D: Rise in intracellular Ca²⁺ in response to electrical stimulus shown as changes in Fura-2 fluorescence intensity (ΔFFI); E: Single exponential intracellular Ca²⁺ decay rate; F: Representative gel blots depicting levels of SERCA2a, Na⁺-Ca²⁺ exchanger (NCX), total/phosphorylated phospholamban (pPLB) and α-tubulin (loading control) using specific antibodies; G: SERCA2a expression; H: NCX expression; and I: pPLB (Ser¹⁶)-to-PLB ratio. Mean±SEM, n = 44–47 cells from 3 mice (panels A-E); or denoted in the graphs (panels F-I); *p<0.05 (two-way ANOVA).</p

Levels of total and phosphorylated Akt, AMPK, ACC and GSK-3β in myocardium from low fat (LF) or high fat (HF)-fed C57 mice with or without TUDCA treatment (300 mg/kg for 15 days).

<p>A: Representative gel blots of Akt, pAkt, AMPK, pAMPK, ACC, pACC, GSK-3β, pGSK-3β and α-tubulin (loading control) using specific antibodies. B: pAkt-to-Akt ratio; C: pAMPK-to-AMPK ratio; D: pACC-to-ACC ratio; and E: pGSK-3β-to-GSK-3β ratio; Meanυ±SEM; sample sizes are denoted in the bar graphs; *p<0.05 (two-way ANOVA).</p

Levels of insulin signaling cascades in myocardium from low fat (LF) or high fat (HF)-fed C57 mice with or without TUDCA treatment (300 mg/kg for 15 days).

<p>A: pIRS-1-to-IRS-1 ratio; B: pJNK-to-JNK ratio; C: pcJun-to-cJun ratio; and D: pERK-to-ERK ratio. Insets: Representative gel blots of total and phosphorylated IRS-1, JNK, cJun and ERK using specific antibodies. α-tubulin was used as the loading control. Mean±SEM; sample sizes are denoted in the bar graphs; *p<0.05 (two-way ANOVA).</p

Cardiomyocyte contractile function at 25°C in low fat (LF) or high fat (HF)-fed C57 mice with or without TUDCA treatment (300 mg/kg for 15 days).

<p>A: Representative traces depicting cell shortening in LF and HF-fed mice; B: Representative traces depicting cell shortening in LF and HF-fed mice with TUDCA treatment; C: Resting cell length; D: Peak shortening (PS, normalized to resting cell length); E: Maximal velocity of shortening (+dL/dt); F: Maximal velocity of relengthening (−dL/dt); G: Time-to-PS (TPS); and I: Time-to-90% relengthening (TR₉₀). Mean±SEM, n = 60–64 cells from 3 mice per group, *p<0.05 (two-way ANOVA).</p

Expression of ER stress proteins in myocardium from low fat (LF) or high fat (HF)-fed C57 mice with or without TUDCA treatment (300 mg/kg for 15 days).

<p>A: Representative gel blots depicting levels of GRP78/BiP, pPERK, PERK, CHOP, eIF2α, peIF2α and α-tubulin (as loading control) using specific antibodies; B: GRP78/Bip expression; C: pPERK-to-PERK ratio; D: pIRE-1 level; E: CHOP expression; and F: peIF2α-to-eIF2α ratio. Mean±SEM; sample sizes are denoted in the bar graphs, *p<0.05 (two-way ANOVA).</p

Correction: Tauroursodeoxycholic Acid Mitigates High Fat Diet-Induced Cardiomyocyte Contractile and Intracellular Ca²⁺ Anomalies

Correction: Tauroursodeoxycholic Acid Mitigates High Fat Diet-Induced Cardiomyocyte Contractile and Intracellular Ca²⁺ Anomalie

Regulation of ECM-related gene expressions following PO.

<p>Analysis of myocardial collagen, MMP, and TIMP expression from Ad KO or wt C57BL/6 mice 2 or 4 weeks following MTAB surgery. (A&B) Quantification of PO fold over sham changes in myocardial mRNA expressions represented as average C(t) value fold sham, where sham is set to 1. (C) Graphical representation of fold over sham mRNA change of collagens (up: blue, down: brown), MMPs (up: green, down: purple), and TIMPs (up: teal, down: orange). (C&D) Quantification of AdKO versus wt changes in myocardial mRNA expressions after PO represented as average C(t) value fold sham, where sham is set to 1. All values are average of n = 4 to 6 mice per group. * = p < 0.05.</p

Adiponectin increases collagen synthesis and secretion, and MMP2 activation from cardiac fibroblasts.

<p>Adiponectin stimulation of isolated neonatal cardiac fibroblasts was followed by analysis of: (A) Intracellular pro-collagen synthesis was assessed by ³H-proline incorporation following adiponectin treatment (5 μg/mL) for 6, 24 or 48 h. Data represent mean values ± SEM from n = 3 experiments using 3 wells per group for quantification. Total secreted collagen was measured in fibroblast conditioned media following adiponectin treatment for 6, 24 or 48 h by picrosirius red staining. MMP2 activation was analyzed by gelatin zymography in conditioned media collected from 6, 24 and 48 h adiponectin treated fibroblasts. Data represented as mean arbitrary units ± SEM from n = 7 experiments. (B) Immunofluorescent images of intracellular collagen I (red) and collagen III (green) synthesized in cardiac fibroblasts at 60x magnification. Cells were treated with adiponectin for 6, 24 and 48 h. Representative images from n = 3 experiments are shown. (C) Immunofluorescent images of extracellular collagen I (green) and collagen III (red) secreted from cardiac fibroblasts at 60x magnification. Cells were treated with adiponectin for 3 days. Cell nuclei were also stained with DAPI (blue). Representative images from n = 3 experiments are shown. Below, 3-dimensional stacks of Ad-treated NCFs immunostained for Collagen I and Collagen III were rotated to show relative vertical orientation of nuclei (DAPI) and collagen (green). Arrow head indicates coverslip. (D) Fibroblast proliferation was assessed by ³H-thymidine incorporation following adiponectin treatment for 6, 24, or 48 h. Data represent mean values ± SEM from n = 3 experiments using 3 wells per group for quantification. (E) Fibroblast migration was assessed using the wound-scratch assay following adiponectin treatment for 6 or 24 h. Cell nuclei were stained with DAPI and imaged using confocal microscopy under a 20x objective. Data represent mean values ± SEM from n = 3 experiments using 7–10 images per group for quantification. Representative images are shown in (F). (G) Western blot analysis of cardiac fibroblast cell lysates treated with adiponectin (5 μg/mL) and/or pre-treated with AngII (1 μM). (H) Immunofluorescent staining for αSMA in cardiac fibroblasts treated with adiponectin and/or pre-treated with AngII (1 μM), and imaged using confocal microscopy under a 20x objective.</p

Adiponectin deficiency is associated with increased basal cardiac fibrosis.

<p>Representative scanning electron micrographs of fixed left ventricular samples from Ad KO or wt C57BL/6 mice 2 or 4 weeks following sham or MTAB surgery. Magnifications shown are x1K, 2K or 5K as indicated. Images shown are representative of 5–10 images of n = 4 to 6 mice per group.</p

Quantitative analysis of scanning electron microscopy and histology studies.

<p>Quantitative analysis of collagen fibre diameter (A&B) and vimentin staining (C&D) was assessed as described in methods. (A&B) Five images were taken per mounted tissue and three tissue samples were produced per animal heart (= 15 images/animal heart). (C&D) Five images were taken per stained slide and three slides were produced per animal heart (= 15 images/animal heart). Mean values and standard deviations were calculated form the values obtained for three different animals per experimental condition (n = 3) and values are mean ± SEM.</p