| Goose fatty liver is one of the world’s three major cuisine. Overfeeding is the most important method to obtain the goose fatty liver. After overfeeding, the fatty liver contain a lot of healthy elements, for example, unsaturated fatty acid, lecithin, microelement et al. It has a high economic value. In the past, there were amount of study on fatty acid in livers, but a little report about cholesterol metabolism in livers, let alone theeffect on overfeeding goose about cholesterol metabolism. HMGCS1 gene regulate the HMG-CoA synthase, which is the essential anzyme for cholesterol biosynthesis. The enzyme HMG-CoA synthase catalyses the condensation of acetoacetyl-CoA and acetyl-CoAto form HMG-CoA plus free CoA. The gene is closed with cholesterol biosynthesis.This experiment was conducted to study the HMGCS1 gene, and obtain the cDNA sequence. Observing the cholesterol biosynthesis relevance to the gene in the overfeeding goose liver. Amplified by the method of RACE to get the purpose gene cDNA sequence. Observe morphological and tissue slice of the overfeeding goose, and analyze the slaughter trait about body weight, liver weight and liver body weight et al. We choosed a cholesterol kit to detect the cholesterol level in goose liver. On the impact of molecular biology technology, we used fluorescence quantitative PCR technique to detect HMGCS1 mRNA expression. In vitro, we changed the semi-situ perfusion method, successful obtaining goose primary hepatocyte and observing the cell dynamically change. To explore the influence of fatty acid, glucose and insulin on fatty deposits in goose primary hepatocyte. The results are as follows:1. We got nucleotide sequencing of a full-length 2947 bp cDNA for the HMG-CoA synthase, and the transcript encoded a protein of 522 amino acids which started an open reading frame, which was submitted to GenBank(Accession No. KF425515). The gene is highly conserved in the evolutionary process, the sequence of goose is most close to duck, and they are belong to the same branch.2. Quantitative results showed that the gene expression in liver was extremely higher than other organizations (P<0.01). The results indicate that the gene expression extremely decreased in overfeeding goose (P<0.01). There is a positive correlation between the cholesterol levels and the activity of the enzyme of the cholesterol biosynthetic pathway. The results can be used to goose breeding and producting goose fat liver.3. After overdeeding the body weight, liver weight and liver body weight was increasing rapidly (P<0.05). Goose liver present fatty liver obviously. Cell swollen and fatty degeneration was happened in hepatocyte, but no inflammatory reaction occurred. On the other hand, even though cholesterol content in serum increased sharaply (P<0.01) in overfeeding goose, but in liver the data was decreased rapidly (P<0.01).4. Using the changed semi-situ perfusion method, successfully obtaining survivable goose primary hepatocyte. It was growing well on 24 wells plate. The best handling time was 48 h to 72 h. With the multiplication of hepatocyte, the lipid droplet became more in intracellular. It is indicate that the goose hepatocyte has a strong metabolism ability, maybe it is the reason that goose own efficient overfeeding. Meanwhile, unsaturated fatty acid and insulin can promote the synthesis of fat in liver cells under certain concentration. At the same time, polyunsaturated fatty acid can also promote the expression of HMGCS1 gene in certain concentration. |
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