CYP-450 Epoxygenase Derived Epoxyeicosatrienoic Acid Contribute To Reversal of Heart Failure in Obesity-Induced Diabetic Cardiomyopathy via PGC-1α Activation

Figures

• 

  Figure 1. EET-agonist treatment improves left ventricular dilation, fractional shortening, and glucose tolerance. (A) Sample M-mode short axis echocardiographic images showing LV dilation and FS in wild-type (C57) and db/db control, db/db non-target and vehicle (saline) injected, db/db treated with EET-agonist, and PGC-1α-deficient db/db mice treated with EET-agonist; (B) Graph depicting left ventricular fractional shortening in each cohort (n=6, **p<0.05 vs. C57, *p<0.05 vs. control and #p<0.05 vs. EET-agonist). Results are mean ± SE, n=6, *p<0.05 vs. db/db control, #p<0.05 vs. db/db mice treated with EET-agonist. Representative blots; (C) and densitometric analysis of (D) pAMPKα/AMPKα, and (E) pIR972, representative blots (F) and densitometric analysis of (G) HO-1, (H) PGC-1α, and (I) Fis1 in WT (C57), db/db control and db/db non-target and vehicle (saline) injected mice. Results are mean ± SE, n=6, *p<0.05 vs. C57 mice.

• 

  Figure 2. Expression of NOV in different tissues of control db/db mice. (A) NOV mRNA expression in pericardial fat and visceral fat; (B) NOV mRNA expression in heart, kidney and liver; (C) Effect of EET-agonist treatment on NOV mRNA expression levels in heart of individual mouse, Effect of EET-agonist treatment on NOV mRNA expression in (D) Pericardial fat and (E) Visceral fat of db/db control mice, db/db treated with EET-agonist mice, and PGC-1α-deficient db/db mice treated with EET-agonist. Results are mean ± SE, n=6, *p<0.05 vs. db/db control, #p<0.05 vs. db/db mice treated with EET-agonist.

• 

  Figure 3. Effect of EET-agonist treatment on levels of PGC-1α, HO-1, NOV Sirt1, pAMPKα, total AMPKα, pAKT, total AKT and pIR972 in cardiac tissue of db/db mice. (A) PGC-1α mRNA levels in control (WT) and lentivirus-mediated knockdown of PGC-1α in 3T3-L1-derived adipocytes. Representative western blots; (B) and G) and densitometric analysis of (C) PGC-1α, (D) HO-1, (E) HO-activity as measured by CO generation, (F) NOV, (H) Sirt1, (I) pAMPKα/AMPKα, (J) pAKT/AKT, and (K) pIR972 in cardiac tissues of db/db control mice, db/db mice treated with EET-agonist, and PGC-1α-deficient db/db mice treated with EET-agonist. Results are mean ± SE, n=6, *p<0.05 vs. db/db control, #p<0.05 vs. db/db mice treated with EET-agonist.

• 

  Figure 4. Effect of EET-agonist treatment on mitochondrial function and biogenesis in cardiac tissue of db/db mice. (A) Representative western blots, densitometric analysis of (B) Mfn2; (C) Fis1, and (D) DRP1, (E) ATP levels, (F) mitochondrial copy number; (G) mRNA expression of an antioxidant gene MnSOD2 in cardiac tissues of db/db control mice, db/db mice treated with EET-agonist, and PGC-1α-deficient db/db mice treated with EET-agonist. Results are mean ± SE, n=6, *p<0.05 vs. db/db control, #p<0.05 vs. db/db mice treated with EET-agonist.

• 

  Figure 5. Effect of EET-agonist treatment on inflammatory cytokine and MMP2 levels in cardiac tissue of db/db mice. Representative blots (A) and densitometric analysis of (B) MMP2, (C) IL-6, and (D) mRNA expression of TNF-α in cardiac tissue of db/db control mice, db/db mice treated with EET-agonist, and PGC-1α-deficient db/db mice treated with EET-agonist. Results are mean ± SE, n=6, *p<0.05 vs. db/db control, #p<0.05 vs. db/db mice treated with EET-agonist.

• 

  Figure 6. Effect of EET-agonist treatment on levels of PGC-1α, HO-1, COX-I and PRDM16, adiponectin, TNF-α, IL-6, and CCL2 in pericardial adipose tissues of db/db mice. mRNA levels of (A) PGC-1α, (B) HO-1, (C) COX-I, (D) PRDM16, (E) Adiponectin, (F) TNF-α, (G) IL-6, and (H) CCL2. Results are mean ± SE, n=6, *p<0.05 vs. db/db control, #p<0.05 vs. db/db mice treated with EET-agonist.

• 

  Figure 7. Schematic description of the EET mediated NOV-PGC-1α-HO-1 in mitochondrial biogenesis and function, and thermogenic genes PRDM16. EET-mediated change of pericardial fat phenotype and increase in to expression of mitochondrial signaling protein, thermogenic genes similar to that described to beige type like adipose tissue. The EET-agonist-mediated reduction in NOV subsequently increases PGC-1α and HO-1 leads to increased ATP production, oxygen consumption, increased mitochondrial fusion, DNA copy number, insulin sensitivity that all in concert leads to an improvement in cardiac function.