| The disorder of Lipid metabolism could lead to the massive triglycerides (TG) accumulation in the liver,then result in hepatic steatosis.At present,the molecular mechanism of hepatic steatosis caused by lipid metabolism disorder in goose hepatocytes has not been reported.In this study,DGAT1,DGAT2,FOXO1, MTP,PLIN and CPT-1 partial sequences of goose were cloned for the first time, which are related with goose lipid metabolism balance.Then the hepatic steatosis model in vitro was constructed by culturing the goose primary hepatocytes with different concentration oleic acid to induce goose hepatocytic steatosis.TG concentration intracellular and extracellular of steatosis hepatocytes,and the change of apoB concentration were measured.Meanwhile,the mRNA expression of DGAT1,DGAT2,PPARα,PPARγ,FOXO1,MTP,PLIN and CPT-1 was dedected by Real-Time RT-PCR.The results were as follows:(1) The partial CDS of MTP,DGAT1,DGAT2,CPT-1,FOXO1 and PLIN genes were amplificationed for the first time,and the length of their cloned sequences were 663bp,874bp,409bp,551bp,1087bp and 277bp, respectively.(2) Gene sequence of MTP,DGAT1,DGAT2,CPT-1,FOXO1 and PLIN of goose has high homology with Gallus gallus or Poephila castanotis above 90%,with other vertebrate by 70%-80%through sequence BLAST.(3) Compared with control group,0.5,1.0 and 1.5 mmol/L oleic oil cultured with hapetocytes for 24h all could increase intracellular TG accumulation by 3 (P＜0.01),3.43(P＜0.01),2.86 times(P＜0.05),respectively.(4) The sesult of Oil Red O staining showed,the lipid droplets in hepatocytes of control group were much less than that in experimental group,and the degree of lipid droplet accumulation increased intra hepatocytes with the increasing of oleic oil concentration.(5) The MTT assay was used for the quantification of cytoactive.After 24h cultured with oleic oil,1.5 mmol/L(P＜0.01) oleic oil had an inhibiting effect on the cytoactive,0.5 mmol/L(P＜0.01)oleic oil could induce cytoactive,and 1.0 mmol/L oleic oil had no effect on cell viability.(6) Compared with control group,0.5,1.0 and 1.5 mmol/L oleic oil cultured with goose hepatocytes for 24h could all decreased secretion of apoB-containing lipoproteins by 0.96,0.83(P＜0.05) and 0.72 times(P＜0.01),respectively.The extracellular TG concentration decreased by 4.69(P＜0.01),2(P＜0.05) and 1.3 times,gradually.(7) Different concentration of oleic oil cultured with hepatocytes for 24h had evident effect on some genes expression related to lipid metabolism. Compared with control group,the mRNA level of PPARγin the experimental groups which were treated with 0.5,1.0 and 1.5 mmol/L oleic oil was increased by 1.893(P＜0.05),1.297 and 0.909 times,respectively;The mRNA level of PLIN was increased by 5.706(P＜0.01),5.31(P＜0.01) and 4.145 times (P＜0.01),respectively.The mRNA level of FOXO1 was decreased by 0.593(P＜0.05),0.402(P＜0.01) and 0.314 times(P＜0.01),respectively; The mRNA level of MTP was decreased by 0.913,0.806(P＜0.05) and 0.543 times(P＜0.01),respectively;The mRNA level of PPARαwas increased by 2.26(P＜0.05),2.957(P＜0.01) and 10.302 times(P＜0.01),respectively; The mRNA level of CPT-1 was increased by 2.107(P＜0.01),2.033(P＜0.01) and 1.631 times(P＜0.05),respectively.The mRNA level of DGAT1 was increased by 1.531(P＜0.05),1.244 and 0.936 times(P＜0.05),respectively. The mRNA level of DGAT2 was increased by 2.651(P＜0.01),2.054 (P＜0.01) and 1.814(P＜0.05) times(P＜0.05),respectively.
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