Effect Of Different Iron Loads On The Hepcidin MRNA Expression In Piglet Liver

In this experiment,piglet models of iron-deficient and iron-overloaded were established by injecting different doses of iron-detran at the age of 3 days, and Hepcidin gene of Sus scrofa was isolated and cloned. Effect of different iron loads on the Hepcidin mRNA abundance in piglet liver was studied with fluorescence quantitative chain reaction (FQ-PCR).The piglet models of iron-deficient and iron-overloaded were established by injecting different doses of iron-detran at the age of 3 days. Iron contents in serum and tissues of liver and spleen were analyzed by Z-2000 atom absorption spectrometer, and the serum biochemical indexes and the antioxidant system were also determined. The results showed that the iron levels in organs and serum were increased (P < 0.05) in the iron-overloaded group, while decreased (P < 0.05) in the liver of the iron-deficient group, 4.7 and 188 times lower in the spleen and serum than that in the control group (P< 0.01). The contents of TP, TB, HDL-C, MDA, GSH-Px and POD in serum were obviously increased (P < 0.05) in the iron-overloaded group, and decreased (P < 0.05) in the iron-deficient group. The levels of AKP and TG were decreased (P < 0.05), compared with the control group.Effect of different iron loads on the Hepcidin mRNA abundance in piglet liver was studied with FQ-PCR. The result demonstrated that Hepcidin mRNA abundance was 131 times lower in the liver of piglet with iron-deficient,while 7 times higher in the iron-overloaded group than that in the control group (P < 0.01).The mRNA from Sus scrofa has been extracted successfully. A 278bp Hepcidin gene was amplified by PCR, and then the PMD19-T vector and 278bp DNA were ligated. PCR detection and digestion with restriction endonucleases of Xbaâ… and Kpnâ… were used to check the inserted DNA.The nucleotide sequence of Hepcidin gene was analyzed, and the phylogenetic tree of Hepcidin amino acid sequence was established. The homology of the gene nucleotide sequences between the amplified gene products and the target sequence of Hepcidin was 100%. Forecast for protein molecular weight and isoelectric points were 8.76kDa and 9.06, respectively. Homology of Hepcidin encoding region is 81.71% between Sus scrofa and Bos Taurus. The phylogenetic tree conformed to the laws of species evolution.