花生四烯酸对日本沼虾肝胰腺细胞脂质代谢基因表达的影响

发布时间：2018-08-03 08:56

【摘要】：本试验旨在评价细胞培养液中花生四烯酸(arachidonic acid,ARA)浓度对日本沼虾肝胰腺细胞活力及脂质代谢相关基因表达的影响。分离日本沼虾肝胰腺细胞,使用M199完全培养液培养5 d后换成含ARA的培养液,ARA浓度分别为0(ARA1)、50(ARA2)、100(ARA3)、200(ARA4)和1 000μmol/L(ARA5),测定12和24 h时脂质代谢相关基因的表达水平,以及24h时细胞活力。结果表明:原代肝胰腺细胞使用完全培养液时,生长状况良好,能存活15 d左右;ARA5组24 h时细胞活力显著低于ARA1和ARA2组(P0.05);高浓度的ARA降低了12和24 h时Δ4脱饱和酶(Δ4 FAD)、Δ6脱饱和酶(Δ6 FAD)、碳链延长酶6(Elovl6)、B类Ⅰ型清道夫受体(SR-BⅠ)、脂肪酸结合蛋白10(FABP10)、乙酰辅酶A结合蛋白(ACBP)基因表达水平;ARA作用12 h时,ARA2组SR-BⅠ基因表达水平显著高于其余各组(P0.05),ARA2和ARA3组FABP10基因表达水平显著高于ARA1和ARA5组(P0.05),ARA3组ACBP基因表达水平显著高于其余各组(P0.05);ARA作用24 h时,ARA2组SR-BⅠ、FABP10和ACBP基因表达水平显著高于其余各组(P0.05)。由此可见,细胞培养液中ARA浓度会影响日本沼虾肝胰腺细胞活力及脂质代谢相关基因的表达,过高的ARA浓度(1 000μmol/L)会降低细胞的活力,适宜的ARA浓度(50~100μmol/L)可促进脂肪酸脱饱和酶、碳链延长酶及脂肪酸转运相关基因的表达。
[Abstract]:The purpose of this study was to evaluate the effects of arachidonic acidoic acid (arachidonic) concentration on hepatopancreatic cell viability and expression of genes related to lipid metabolism of Macrobrachium japonicus. The hepatopancreatic cells of Macrobrachium japonicus were isolated and cultured in M199 for 5 days. The concentrations of ARA were 0 (ARA1) ARA2 (ARA3) 200 (ARA4) and 1 000 渭 mol/L (ARA5) respectively. The expression levels of lipid metabolism-related genes at 12 and 24 h and the cell viability at 24 h were measured. The results showed that the primary hepatopancreatic cells grew well when they were treated with complete culture medium. The cell viability of ARA5 group was significantly lower than that of ARA1 and ARA2 group at 24 h after survival for 15 d (P0.05), and that of high concentration ARA decreased 螖 4 FAD), 螖 6 desaturase (螖 6 FAD), C chain extendease 6 (Elovl6) SR-B 鈪，

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