Berdeaux O, Blond J P, Bretillon L, Chardigny J M, Mairot T, Vatèle J M, Poullain D, Sébédio J L
INRA, Unité de Nutrition Lipidique, Dijon, France.
Mol Cell Biochem. 1998 Aug;185(1-2):17-25. doi: 10.1023/a:1006859616647.
Several nutritional studies have shown the in vivo conversion of the 9c, 12t-18:2 and 9t, 12c-18:2 into long chain polyunsaturated fatty acids (PUFA) containing 20 carbons (geometrical isomers of eicosadienoic and eicosatetraenoic acids). In the present work, some in vitro studies were carried out in order to have precise information on the conversion of these two isomers. In a first set of experiments, studies were focused on the in vitro delta6 desaturation, the first regulatory step of the biosynthesis of n-6 long chain PUFA, from 9c, 12c-18:2. Rat liver microsomes were prepared and incubated under desaturation conditions with [1-14C]-9c, 12c-18:2 in presence of unlabelled 9c, 12t-, 9t, 12c- or 9t, 12t-18:2. The data show that each trans isomer induced a decrease of the delta6 desaturation of the [1-14C]-9c, 12c-18:2, but the 9c, 12t-18:2 was the most potent inhibitor (up to 63%). Rat liver microsomes were also incubated with [1-14C]-9c, 12c-18:2, [1-14C]-9c, 12t-18:2 or [1-14C]-9t, 12c-18:2 under desaturation conditions. The results indicated that 18:2 delta9c, 12t is a much better substrate for desaturase than 9t, 12c-18:2. Moreover, the conversion levels of [1-14C]-9c, 12t-18:2 was similar to what was observed for its all cis homologue, at low substrate concentration only. In a second set of experiments, in vitro elongation studies of each mono-trans 18:2 isomer and 9c, 12c-18:2 were carried out. For that purpose, rat liver microsomes were incubated with [1-14C]-9c, 12c-18:2, [1-14C]-9c, 12t-18:2 or [1-14C]-9t, 12c-18:2 underelongation conditions. The data show that [1-14C]-9t, 12c-18:2 is betterelongated than 9c, 12c-18:2 while the amount of product formed from [1-14C]-9c, 12t-18:2 was lower than was produced from the 9c, 12c-18:2. Thus, the desaturation enzymes presented a higher affinity for the 9c, 12t-18:2 whereas the elongation enzyme presented a higher affinity for the 9t, 12c-18:2.
多项营养研究表明,9顺式,12反式-18:2和9反式,12顺式-18:2在体内可转化为含20个碳的长链多不饱和脂肪酸(PUFA)(二十碳二烯酸和二十碳四烯酸的几何异构体)。在本研究中,进行了一些体外研究,以便获得有关这两种异构体转化的精确信息。在第一组实验中,研究集中在体外Δ6去饱和作用,这是n-6长链PUFA生物合成的第一个调控步骤,底物为9顺式,12顺式-18:2。制备大鼠肝脏微粒体,并在去饱和条件下,与[1-14C]-9顺式,12顺式-18:2一起,在未标记的9顺式,12反式-、9反式,12顺式-或9反式,12反式-18:2存在的情况下进行孵育。数据表明,每种反式异构体均会导致[1-14C]-9顺式,12顺式-18:2的Δ6去饱和作用降低,但9顺式,12反式-18:2是最有效的抑制剂(高达63%)。大鼠肝脏微粒体也在去饱和条件下与[1-14C]-9顺式,12顺式-18:2、[1-14C]-9顺式,12反式-18:2或[1-14C]-9反式,12顺式-18:2一起孵育。结果表明,18:2Δ9顺式,12反式比9反式,12顺式-18:2是去饱和酶更好的底物。此外,[1-14C]-9顺式,12反式-18:2的转化水平仅在低底物浓度下与其全顺式同系物的观察结果相似。在第二组实验中,对每种单反式18:2异构体和9顺式,12顺式-18:2进行了体外延伸研究。为此,将大鼠肝脏微粒体在延伸条件下与[1-14C]-9顺式,12顺式-18:2、[1-14C]-9顺式,12反式-18:2或[1-14C]-9反式,12顺式-18:2一起孵育。数据表明,[1-14C]-9反式,12顺式-18:2比9顺式,12顺式-18:2更容易延伸,而由[1-14C]-9顺式,12反式-18:2形成的产物量低于由9顺式,12顺式-18:2产生的量。因此,去饱和酶对9顺式,12反式-18:2具有更高的亲和力,而延伸酶对9反式,12顺式-18:2具有更高的亲和力。