Nathans A H, Kitabchi A E
Biochim Biophys Acta. 1975 Aug 13;399(2):244-53. doi: 10.1016/0304-4165(75)90255-x.
Isolated adrenal cells from Vitamin E-deficient and control rats were prepared by a trypsin digestion method. Cyclic adenosine 3',5'-monophosphate (cyclic AMP) formation was studied in response to adrenocorticotropin (ACTH) in the presence and absence of ascorbate by measuring the conversion of prelabeled adenosine 5'-triphosphate [14C]ATP to cyclic [14C]AMP. Ascorbate (0.5 mM) inhibited ACTH-induced cyclic [14C]AMP formation in adrenal cells isolated from Vitamin E-deficient rats but had no effect in the control cells. The inhibitory effect of ascorbate on ACTH-induced cyclic AMP formation in Vitamin E-deficient rats decreased as the concentration of ACTH increased. In Vitamin E-deficient rats ascorbate inhibited ACTH-induced cyclic [14C]AMP formation after 30 min of incubation. There was no further significant accumulation of cyclic [14C]AMP at 60 min or 120 min although in the absence of ascorbate cyclic [14C]AMP continued to be formed. The in vitro addition of alpha-tocopherol reduced the inhibition of ACTH-induced cyclic [14C]AMP formation by ascorbate in Vitamin E-deficient rats. These studies suggest that alpha-tocopherol and ascorbate may affect ACTH-induced cyclic AMP formation through interaction with the membrane-bound enzyme adenylate cyclase.
采用胰蛋白酶消化法制备了维生素E缺乏大鼠和对照大鼠的分离肾上腺细胞。通过测量预标记的腺苷5'-三磷酸[14C]ATP向环[14C]AMP的转化，研究了在有和没有抗坏血酸的情况下，肾上腺皮质激素（ACTH）刺激下环腺苷3',5'-单磷酸（环AMP）的生成。抗坏血酸（0.5 mM）抑制了维生素E缺乏大鼠分离的肾上腺细胞中ACTH诱导的环[14C]AMP生成，但对对照细胞没有影响。随着ACTH浓度的增加，抗坏血酸对维生素E缺乏大鼠中ACTH诱导的环AMP生成的抑制作用减弱。在维生素E缺乏大鼠中，孵育30分钟后抗坏血酸抑制了ACTH诱导的环[14C]AMP生成。尽管在没有抗坏血酸的情况下环[14C]AMP继续生成，但在60分钟或120分钟时环[14C]AMP没有进一步显著积累。在维生素E缺乏大鼠中，体外添加α-生育酚可降低抗坏血酸对ACTH诱导的环[14C]AMP生成的抑制作用。这些研究表明，α-生育酚和抗坏血酸可能通过与膜结合酶腺苷酸环化酶相互作用来影响ACTH诱导的环AMP生成。
