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

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。

Effects of Arachidonic Acid on Lipid Metabolism-Related Gene Expressions of Hepatopancreas Cells Isolated from Juvenile Oriental River Prawn,Macrobrachium nipponense

This experiment was conducted to determine the effects of arachidonic acid (ARA) concentration in culture medium on cell viability and lipid metabolism-related gene expressions of hepatopancreas cells isolated from juvenile oriental river prawn, Macrobrachium nipponense.The hepatopancreas cells were dissected from prawns and were cultured with complete culture medium for 5 days.After that, cultured cells were incubated in medium supplemented with graded levels 0 (ARA1) , 50 (ARA2), 100 (ARA3) , 200 (ARA4) and1 000 μmol/L (ARA5) of ARA.Cell viability at 24 h and gene expressions of lipid metabolism-related genes at 12 and 24 h were examined.The results showed as follows: the hepatopancreas cells showed well growth in complete culture medium, and could survive for 15 days;cell viability was significantly decreased by incubation with higher level (1 000 μmol/L) of ARA (ARA5 group) compared with ARA1 and ARA2 groups (P<0.05) after 24 h;higher level (1 000 μmol/L) of ARA (ARA5 group) caused significant decreases of gene expressions of delta-4 fatty acyl desaturase (Δ4 FAD), delta-6 fatty acyl desaturase (Δ6 FAD), very-longchain fatty acids-6 (Elovl6), scavenger receptor class B type Ⅰ (SR-B Ⅰ), fatty acid-binding protein 10(FABP10) and acyl-CoA binding protein (ACBP) of hepatopancreas cells incubation for both 12 and 24 h;after incubation with ARA for 12 h, the gene expression of SR-B Ⅰ of ARA2 group was significantly higher than that of other groups (P<0.05), FABP10 gene expression of ARA2 and ARA3 groups was significantly higher than that of ARA1 and ARA5 groups (P<0.05), and ACBP gene expression of ARA3 group was significantly higher than that of other groups (P<0.05);after incubation with ARA for 24 h, the highest expressions of SR-B Ⅰ , FABP10 and ACBP were observed in ARA2 group, which was significantly higher than those of other groups (P<0.05).These findings suggest that ARA can influence cell viability and lipid metabolism-related gene expressions of hepatopancreas cell isolated from Macrobrachium nipponense.Cell viability can be decreased by incubation with higher level of ARA (1 000 μmol/L).Appropriate levels of ARA (50 to100 μmol/L) can promote the expressions of genes related to fatty acyl desaturase, elongases of very-longchain fatty acids and fatty acid transport.Chinese Journal of Animal Nutrition, 2017, 29 (2) : 536-546]