Korean J Physiol Pharmacol.  2016 Sep;20(5):477-485. 10.4196/kjpp.2016.20.5.477.

Histone deacetylase inhibitor, CG200745, attenuates cardiac hypertrophy and fibrosis in DOCA-induced hypertensive rats

• Lee E
• Song MJ
• Lee HA
• Kang SH
• Kim M
• Yang EK
• Lee DY
• Ro S
• Cho JM
• Kim I

• KMID: 2350504
• DOI: http://doi.org/10.4196/kjpp.2016.20.5.477

Abstract

CG200745 is a novel inhibitor of histone deacetylases (HDACs), initially developed for treatment of various hematological and solid cancers. Because it is water-soluble, it can be administered orally. We hypothesized that the HDAC inhibitor, CG200745, attenuates cardiac hypertrophy and fibrosis in deoxycorticosterone acetate (DOCA)-induced hypertensive rats. For establishment of hypertension, 40 mg/kg of DOCA was subcutaneously injected four times weekly into Sprague-Dawley rats. All the rats used in this study including those in the sham group had been unilaterally nephrectomized and allowed free access to drinking water containing 1% NaCl. Systolic blood pressure was measured by the tail-cuff method. Blood chemistry including sodium, potassium, glucose, triglyceride, and cholesterol levels was analyzed. Sections of the heart were visualized after trichrome and hematoxylin and eosin stain. The expression of hypertrophic genes such as atrial natriuretic peptide A (Nppa) and atrial natriuretic peptide B (Nppb) in addition to fibrotic genes such as Collagen-1, Collagen-3, connective tissue growth factor (Ctgf), and Fibronectin were measured by quantitative real-time PCR (qRT-PCR). Injection of DOCA increased systolic blood pressure, heart weight, and cardiac fibrosis, which was attenuated by CG200745. Neither DOCA nor CG200745 affected body weight, vascular contraction and relaxation responses, and blood chemistry. Injection of DOCA increased expression of both hypertrophic and fibrotic genes, which was abrogated by CG200745. These results indicate that CG200745 attenuates cardiac hypertrophy and fibrosis in DOCA-induced hypertensive rats.

Keyword

Cardiac fibrosis; Cardiac hypertrophy; CG200745; Deoxycorticosterone acetate (DOCA); Histone deacetylase

MeSH Terms

Animals
Blood Pressure
Body Weight
Cardiomegaly*
Chemistry
Cholesterol
Connective Tissue Growth Factor
Desoxycorticosterone
Desoxycorticosterone Acetate
Drinking Water
Eosine Yellowish-(YS)
Fibronectins
Fibrosis*
Glucose
Heart
Hematoxylin
Histone Deacetylase Inhibitors*
Histone Deacetylases*
Histones*
Hypertension
Methods
Potassium
Rats*
Rats, Sprague-Dawley
Real-Time Polymerase Chain Reaction
Relaxation
Sodium
Triglycerides
Cholesterol
Connective Tissue Growth Factor
Desoxycorticosterone
Drinking Water
Eosine Yellowish-(YS)
Fibronectins
Glucose
Hematoxylin
Histone Deacetylase Inhibitors
Histone Deacetylases
Histones
Potassium
Sodium

Figure

• 

  Fig. 1 Effect of CG200745 on systolic blood pressure and body weight in deoxycorticosterone acetate (DOCA)-induced hypertensive rats.(A) Systolic blood pressures were measured using the tail-cuff method in the sham group (n=6), DOCA alone group (n=6), DOCA plus 1.25 mg/kg of CG200745 (CG) administration group (n=6), and DOCA plus 5.0 mg/kg of CG administration group (n=6) for a period of four weeks. Administration of CG attenuated DOCA-induced hypertension. (B) Body weights were monitored for four weeks. DOCA injection with or without CG administration did not affect body weight gain. Data show the mean ± standard error (SE) of six independent experiments (**p<0.01 vs. sham group, ##p<0.01 vs. DOCA alone group).

• 

  Fig. 2 Effects of CG200745 on aortic ring contraction and relaxation in deoxycorticosterone acetate (DOCA)-induced hypertensive rats.Phenylephrine (PE) was cumulatively added for vascular contraction in the aortic rings with (A) or without (C) endothelium. Developed tension is expressed as a percentage of the maximal contraction to 50 mmol/L KCl. Acetylcholine (ACh) or sodium nitroprusside (SNP) was cumulatively added for vasorelaxation in the aortic rings with (B) or without (D) endothelium, respectively. Developed relaxation is expressed as a percentage of the maximal contraction to PE. Neither DOCA nor CG affected aortic ring contraction and relaxation. Data show the mean±standard error (SE) of six independent experiments.

• 

  Fig. 3 Effect of CG200745 on heart and lung weight in deoxycorticosterone acetate (DOCA)-induced hypertensive rats.Cardiac hypertrophy was analyzed based on heart weight (HW)/tibia length (TL) ratios in the sham group (n=6), DOCA alone group (n=6), DOCA plus 1.25 mg/kg of CG administration group (n=6), and DOCA plus 5.0 mg/kg of CG administration group (n=6). (A, B) HW/TL and left heart weight (LHW)/TL ratios were increased in the DOCA alone group as compared with the sham group. Administration of CG results in restoration of HW/TL and LHW/TL ratios. (C, D) Lung weight (LW) and right heart weight (RHW) were similar among all groups. CG administration did not affect RHW/TL and LW/TL ratios. Data show the mean±standard error (SE) of six independent experiments (**p<0.01 vs. sham group, #p<0.05 vs. DOCA alone group).

• 

  Fig. 4 H&E stain of hearts of deoxycorticosterone acetate (DOCA)-induced hypertensive rats.Analyses of histology of the sham group (n=6), DOCA alone group (n=6), DOCA plus 1.25 mg/kg of CG administration group (n=6), and DOCA plus 5.0 mg/kg of CG administration group (n=6) hearts were performed using hematoxylin and eosin (H&E). The DOCA alone group increased levels of hypertrophy when compared with the sham group as shown by H&E stain, which were attenuated by CG administration. Scale bar is 50 µm.

• 

  Fig. 5 Effect of CG200745 on expression of cardiac hypertrophic genes in deoxycorticosterone acetate (DOCA)-induced hypertensive rats.Expression of atrial natriuretic peptide A (Nppa, A) and atrial natriuretic peptide B (Nppb, B), cardiac hypertrophy markers, were detected by quantitative real-time PCR (qRT-PCR). The DOCA alone group increased expression of Nppa and Nppb mRNA which were decreased with CG administration in the DOCA plus CG administration groups. Data show the mean±standard error (SE) of six independent experiments (*p<0.05 and **p<0.01 vs. sham group, #p<0.05 and ##p<0.01 vs. DOCA alone group).

• 

  Fig. 6 Trichrome stain of hearts of deoxycorticosterone acetate (DOCA)-induced hypertensive rats.Analyses of histology of the sham group (n=6), DOCA alone group (n=6), DOCA plus 1.25 mg/kg of CG administration group (n=6), and DOCA plus 5.0 mg/kg of CG administration group (n=6) hearts were performed using trichrome stain. The DOCA alone group increased cardiac fibrosis (blue stain) when compared with the sham group. CG administration results in attenuated cardiac fibrosis in the DOCA plus CG administration groups. Scale bar is 50 µm.

• 

  Fig. 7 Effect of CG200745 on expression of cardiac fibrotic genes in deoxycorticosterone acetate (DOCA)-induced hypertensive rats.Gene expression of Collagen-1 (A), Collagen-3 (B), connective tissue growth factor (Ctgf, C), Fibronectin (D), and cardiac fibrosis markers were detected by quantitative real-time PCR (qRT-PCR). The DOCA alone group increased expression of collagen-1, collagen-3, Ctgf, and fibronectin mRNA which were abolished by CG administration. Data show the mean±standard error (SE) of six independent experiments (*p<0.05 and **p<0.01 vs. sham group, #p<0.05 and ##p<0.01 vs. DOCA alone group).

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