Korean J Physiol Pharmacol.  2018 Nov;22(6):607-616. 10.4196/kjpp.2018.22.6.607.

The effect of melatonin on cardio fibrosis in juvenile rats with pressure overload and deregulation of HDACs

• Wu Y
• Si F
• Luo L
• Jing F
• Jiang K
• Zhou J
• Yi Q

• KMID: 2430083
• DOI: http://doi.org/10.4196/kjpp.2018.22.6.607

Abstract

The effect of melatonin on juveniles with cardio fibrosis is poorly understood. We investigated whether HDACs participate in the anti-fibrotic processes regulated by melatonin during hypertrophic remodeling. Abdominal aortic constriction (AAC) was employed in juvenile rats resulting in pressure overload-induced ventricular hypertrophy and melatonin was subsequently decreased via continuous light exposure for 5 weeks after surgery. AAC rats displayed an increased cross-sectional area of myocardial fibers and significantly elevated collagen deposition compared to sham-operated rats, as measured by HE and Masson Trichrome staining. Continuous light exposure following surgery exacerbated the increase in the cross-sectional area of myocardial fibers. The expression of HDAC1, HDAC2, HDAC3, HDAC4 and HDAC6 genes were all significantly enhanced in AAC rats with light exposure relative to the other rats. Moreover, the protein level of TNF-α was also upregulated in the AAC light exposure groups when compared with the sham. However, Smad4 protein expression was unchanged in the juveniles' hearts. In contrast, beginning 5 weeks after the operation, the AAC rats were treated with melatonin (10 mg/kg, intraperitoneal injection every evening) or vehicle 4 weeks, and sham rats were given vehicle. The changes in the histological measures of cardio fibrosis and the gene expressions of HDAC1, HDAC2, HDAC3, HDAC4 and HDAC6 were attenuated by melatonin administration. The results reveal that melatonin plays a role in the development of cardio fibrosis and the expression of HDAC1, HDAC2, HDAC3, HDAC4 and HDAC6 in cardiomyocytes.

Keyword

Fibrosis; Heart; Histone deacetylase; Hypertrophy; Juveniles; Melatonin

MeSH Terms

Animals
Collagen
Constriction
Fibrosis*
Gene Expression
Heart
Histone Deacetylases
Hypertrophy
Injections, Intraperitoneal
Melatonin*
Myocytes, Cardiac
Rats*
Smad4 Protein
Collagen
Histone Deacetylases
Melatonin
Smad4 Protein

Figure

• 

  Fig. 1 Effects of AAC on the cardio fibrosis under different light conditions. Representative myocardial HE (A) staining and Masson collagen staining (B). Effects of chronically increased pressure overload on the cross-sectional area of myocardial fibers (C) and collagen deposition (D) under different light conditions. **p<0.01, ***p<0.001 vs. sham group; ###p<0.001 vs. AAC group. Images were acquired at 200×magnification. “n” represents the number of cardiac fibers in Fig. 1C, and the number of image fields in Fig. 1D.

• 

  Fig. 2 Effects of AAC on the cardio fibrosis in melatonin-treated rats. Representative myocardial HE (A) staining and Masson collagen staining (B). Effects of chronically increased pressure overload on the cross-sectional area of myocardial fibers (C) and collagen deposition (D) in melatonin-or vehicle-treated rats. *p<0.05, ***p<0.001 vs. sham group; ###p<0.001 vs. AAC group. Images were acquired at 200×magnification. “n” represents the number of the cardiac fibers in Fig. 2C, and the number of image fields in Fig. 2D.

• 

  Fig. 3 (A-E) Relative HDAC1, 2, 3, 4 and 6 mRNA expression levels in ventricular cardiomyocytes of juvenile rat under different light exposure (real-time PCR). *p<0.05, **p<0.01, ***p<0.001 vs. sham group; ##p<0.01, ###p<0.001, vs. AAC group; all n=8.

• 

  Fig. 4 (A-E) Relative HDAC1, 2, 3, 4 and 6 mRNA expression levels in placebo-treated or melatonin-treated juvenile rat ventricular cardiomyocytes (real-time PCR). **p<0.01, ***p<0.001 vs. Sham group; ##p<0.01, ###p<0.001, vs. AAC group; all n=8.

• 

  Fig. 5 Relative TNF-α and Smad4 protein expression levels in AAC-treated juvenile rat ventricular cardiomyocytes (Western blot). Western blot images (A) and relative quantification of TNF-α (B) and Smad-4 (C) protein expression in juvenile rat ventricular cardiomyocytes. *p<0.05 vs. sham group, all n=4.

• 

  Fig. 6 Relative TNF-α and Smad4 protein expression levels in AAC-treated juvenile rat ventricular cardiomyocytes (Western blot). Western blot images (A) and relative quantification of TNF-α (B) and Smad-4 (C) protein expression in juvenile rat ventricular cardiomyocytes. *p<0.05 vs. sham group, all n=6.

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