【摘要】：本試驗(yàn)旨在評(píng)價(jià)細(xì)胞培養(yǎng)液中花生四烯酸(arachidonic acid,ARA)濃度對(duì)日本沼蝦肝胰腺細(xì)胞活力及脂質(zhì)代謝相關(guān)基因表達(dá)的影響。分離日本沼蝦肝胰腺細(xì)胞,使用M199完全培養(yǎng)液培養(yǎng)5 d后換成含ARA的培養(yǎng)液,ARA濃度分別為0(ARA1)、50(ARA2)、100(ARA3)、200(ARA4)和1 000μmol/L(ARA5),測(cè)定12和24 h時(shí)脂質(zhì)代謝相關(guān)基因的表達(dá)水平,以及24h時(shí)細(xì)胞活力。結(jié)果表明:原代肝胰腺細(xì)胞使用完全培養(yǎng)液時(shí),生長(zhǎng)狀況良好,能存活15 d左右;ARA5組24 h時(shí)細(xì)胞活力顯著低于ARA1和ARA2組(P0.05);高濃度的ARA降低了12和24 h時(shí)Δ4脫飽和酶(Δ4 FAD)、Δ6脫飽和酶(Δ6 FAD)、碳鏈延長(zhǎng)酶6(Elovl6)、B類(lèi)Ⅰ型清道夫受體(SR-BⅠ)、脂肪酸結(jié)合蛋白10(FABP10)、乙酰輔酶A結(jié)合蛋白(ACBP)基因表達(dá)水平;ARA作用12 h時(shí),ARA2組SR-BⅠ基因表達(dá)水平顯著高于其余各組(P0.05),ARA2和ARA3組FABP10基因表達(dá)水平顯著高于ARA1和ARA5組(P0.05),ARA3組ACBP基因表達(dá)水平顯著高于其余各組(P0.05);ARA作用24 h時(shí),ARA2組SR-BⅠ、FABP10和ACBP基因表達(dá)水平顯著高于其余各組(P0.05)。由此可見(jiàn),細(xì)胞培養(yǎng)液中ARA濃度會(huì)影響日本沼蝦肝胰腺細(xì)胞活力及脂質(zhì)代謝相關(guān)基因的表達(dá),過(guò)高的ARA濃度(1 000μmol/L)會(huì)降低細(xì)胞的活力,適宜的ARA濃度(50~100μmol/L)可促進(jìn)脂肪酸脫飽和酶、碳鏈延長(zhǎng)酶及脂肪酸轉(zhuǎn)運(yùn)相關(guān)基因的表達(dá)。
[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，

本文編號(hào)：2161242