Molecular characterisation of tumour necrosis factor alpha and its potential connection with lipoprotein lipase and peroxisome proliferator-activated receptors in blunt snout bream (Megalobrama amblycephala)

J Appl Genet. 2017 Aug;58(3):381-391. doi: 10.1007/s13353-017-0390-9. Epub 2017 Feb 13.

Abstract

Tumour necrosis factor alpha (TNF-α) is one kind of cytokines which is related to inflammation and lipid metabolism. TNF-α cDNA was cloned from the liver of blunt snout bream (Megalobrama amblycephala) through real-time polymerase chain reaction (PCR) and rapid amplification of cDNA ends (RACE) methods. The full-length cDNA of TNF-α covered 1467 bp, with an open reading frame (ORF) of 723 bp, which encodes 240 amino acids. It possessed the TNF family signature IIIPDDGIYFVYSQ. After the lipopolysaccharide (LPS) challenge test, a graded tissue-specific expression pattern of TNF-α was observed and there was high expression abundance in the kidney, brain and liver. After 8 weeks feeding trial, liver samples, two groups fed with 6% and 11% lipid levels, were collected. The results showed that, for fish fed with high-fat diet, the triglyceride of serum and lipid content of liver were elevated. Furthermore, TNF-α and peroxisome proliferator-activated receptors (PPARα, β) mRNA expression of fish fed 11% lipid diet were significantly up-regulated (p < 0.05). Lipoprotein lipase (LPL) and PPARγ mRNA expression of fish fed 11% lipid lever diet were significantly decreased compared to those of fish fed 6% (p < 0.05). The differences between the various expression of related genes in the high and low fat groups demonstrated that TNF-α played a key role in lipid metabolism, which may have an influence on fat metabolism through reducing fat synthesis and strengthening the β-oxidation of fatty acid. These discrepancies warrant further research.

Keywords: Gene expression; Lipoprotein lipase (LPL); Megalobrama amblycephala; Peroxisome proliferator-activated receptors (PPARs); Tumour necrosis factor alpha (TNF-α).

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cloning, Molecular
  • Cyprinidae / metabolism*
  • DNA, Complementary / genetics
  • Diet, High-Fat
  • Lipid Metabolism*
  • Lipoprotein Lipase / metabolism*
  • Liver / metabolism
  • Open Reading Frames
  • Peroxisome Proliferator-Activated Receptors / metabolism*
  • Real-Time Polymerase Chain Reaction
  • Triglycerides / blood
  • Tumor Necrosis Factor-alpha / genetics
  • Tumor Necrosis Factor-alpha / metabolism*

Substances

  • DNA, Complementary
  • Peroxisome Proliferator-Activated Receptors
  • Triglycerides
  • Tumor Necrosis Factor-alpha
  • Lipoprotein Lipase