Porcine sirtuin 1 gene clone, expression pattern, and regulation by resveratrol1

Abstract

Sirtuin1 (Sirt1) is a NAD-dependent deacetylase that plays important roles in a variety of biological processes. In the current study, we examined tissue-specific and different expression pattern of porcine Sirt1 and the effect of resveratrol (RES) on expression of Sirt1 in porcine adipocytes. The full-length complementary DNA sequence of porcine Sirt1 was 4,024 bp (GenBank accession no: EU030283), with a 2,226-bp open reading frame encoding a 742-AA protein (a predicted molecular mass of 80.9 kDa; GenBank accession no. ABS29571). Comparison of the deduced AA sequence with the corresponding sequences of human, dog, cattle, and mouse Sirt1 showed 82 to 92% similarity. Furthermore, the porcine Sirt1 was highly expressed in porcine brain, to a lesser degree in spleen and white adipose tissue, and had low but detectable expression in liver. In subcutaneous adipose tissue and omental adipose tissue, expression of the porcine Sirt1 mRNA was greater in adult pigs than in young pigs (P < 0.01). In vitro, exposure of cultured adipocytes to 40 and 80 micro M RES for 24 h increased mRNA levels of porcine Sirt1 by 47.86% (P < 0.01) and 91.04% (P < 0.01), respectively. Accordingly, lipid accumulation and NEFA release were decreased (P < 0.05), respectively. After cultures were treated with RES for 48 h, the mRNA level of porcine Sirt1 was increased by 103.84% (P < 0.01) and 148.79% (P < 0.01), respectively. Lipid accumulation was decreased and NEFA release was increased (P < 0.05), respectively. These results provide information needed for manipulating Sirt1 expression in regulating fat deposition in pigs.

Effect of breed on the expression of Sirtuins (Sirt1-7) and antioxidant capacity in porcine brain

“…The purity and concentration of total RNA were determined by NanoDrop 2000 (Thermo Scientific Inc., Wilmington, DE, USA). For each sample, 2 μg of total RNA were used to synthesize cDNA according to our procedures published previously (Shan et al, 2009). …”

“…In this study, 18S rRNA (18S) was used as the reference gene. Oligonucleotide primers (Shanghai Sangon Co., Ltd, Shanghai, China) specific for porcine Sirt1-7, and 18S were based on previous studies (Jin et al, 2009;Shan et al, 2009) (Supplementary Table S1). The PCR system and cycling conditions were the same as those described previously (Shan et al, 2009).…”

“…In pigs, high levels of porcine Sirt1-7 mRNA were also found in brain (Shan et al, 2009;Ghinis-Hozumi et al, 2013). At the same time, the antioxidants such as superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) activity in pork could be affected by genetic type (Hernandez et al, 2004).…”

“…Therefore, based on our previous studies (Shan et al, 2009 and, the objective of the current study was to compare the differences of Sirtuins gene expression and the concentrations of the T-AOC, total superoxide dismutase (T-SOD), GSH-Px, CAT, and malondialdehyde (MDA) in brain between Jinhua pigs (a fatty breed of China) and Landrace pigs (a leaner breed) at the same age (180 days of age), as well as the same BW of 64 kg, respectively. In addition, the effects of age on Sirtuins expression in porcine brain were also studied.…”

Potential role of sirtuins in livestock production

“…Finally, an applied study was made available by Weber et al (2010), who evaluated the effect of a high fiber diet on the intestinal tract of growing, female pigs and observed a decrease in liver mass and energy stores and an increase in plasma triglycerides, in addition to higher Sirt1, PGC-1a (peroxisome proliferator-activated receptor-g coactivator) and CoxII (cyclooxygenase-II) expression, suggesting a more efficient energy utilization in this tissue. In summary, Bai et al (2008) and Jin et al (2009) studied porcine sirtuins, focusing on their role as models of human physiology for the discovery of therapeutic treatments; Shan et al (2009 and focused on the potential use of Sirt1 regulation to control fat deposition in pigs, which is higher in adult pigs and lean breeds, and Weber et al (2010) performed the first application, regulating sirtuin expression by dietary manipulations.…”