Vasoactive intestinal peptide induces the synthesis of the cholesterol side-chain cleavage enzyme complex in cultured rat ovarian granulosa cells.
Vasoactive intestinal peptide induces the synthesis of the cholesterol side-chain cleavage enzyme complex in cultured rat ovarian granulosa cells.
复制标题
血管活性肠肽诱导培养的大鼠卵巢颗粒细胞中胆固醇侧链裂解酶复合物的合成。
DOI：
10.1073/pnas.83.19.7490
复制
发表时间：
1986
影响因子：
11.1
通讯作者：
Ojeda,SR
中科院分区：
文献类型：
--
作者：
Trzeciak,WH;Ahmed,CE;Simpson,ER;Ojeda,SR
已在卵巢神经中发现血管活性肠肽 (VIP)，可刺激未成熟卵巢分泌类固醇。为了深入了解其作用机制，在未成熟雌激素引发的大鼠卵巢颗粒细胞中研究了 VIP 对胆固醇侧链裂解酶复合物合成的影响。将细胞在无血清培养基中培养48小时；蛋白质用[35S]蛋氨酸标记；用针对牛肾上腺酶的多克隆抗体进行免疫分离后，通过电泳分析评估细胞色素P-450、铁硫蛋白和NADPH：铁硫蛋白还原酶的合成。浓度范围为 0.001 至 1 µM 的 VIP 可刺激细胞色素 P-450 和铁硫蛋白的合成 3 至 5 倍。肽 NH2 末端组氨酸、COOH 末端异亮氨酸与 VIP 具有大于 50% 的序列同源性，以约 50% VIP 的有效性刺激两种蛋白质的合成。促胰液素是胰高血糖素-促胰液素肽家族的另一个成员，与 VIP 仅有 30% 的序列同源性，但没有效果。使用 NADPH：铁硫蛋白还原酶观察到类似的结果。 VIP 诱导的胆固醇侧链裂解酶复合物的合成伴随着 cAMP 积累和孕激素形成的剂量相关增加。结论是，VIP调节卵巢胆固醇侧链裂解酶复合物的合成，催化孕酮生物合成中的限速反应，并且VIP效应至少部分是通过cAMP介导的。提示VIP对卵巢孕酮合成的刺激作用可能有助于调节卵巢的功能发育。
Vasoactive intestinal peptide (VIP) has been identified in ovarian nerves and stimulates steroid secretion from immature ovaries. To gain insight into its mechanism of action, the effect of VIP on the synthesis of the cholesterol side-chain cleavage enzyme complex was studied in ovarian granulosa cells from immature estrogen-primed rats. The cells were cultured for 48 hr in serum-free medium; the proteins were labeled with [35S]methionine; and the synthesis of cytochrome P-450, iron-sulfur protein, and NADPH:iron-sulfur protein reductase was evaluated by electrophoretic analysis after immunoisolation with polyclonal antibodies directed against the bovine adrenal enzymes. VIP at concentrations ranging from 0.001 to 1 microM stimulated 3- to 5-fold the synthesis of cytochrome P-450 and iron-sulfur protein. Peptide NH2-terminal histidine, COOH-terminal isoleucine, which has greater than 50% sequence homology of VIP, stimulated the synthesis of both proteins at approximately 50% of VIP effectiveness. Secretin, another member of the glucagon-secretin family of peptides, which has only 30% sequence homology to VIP, was without effect. Similar results were observed with the NADPH:iron-sulfur protein reductase. VIP-induced synthesis of the cholesterol side-chain cleavage enzyme complex was accompanied by a dose-related increase in cAMP accumulation and progestin formation. It is concluded that VIP regulates the synthesis of the ovarian cholesterol side-chain cleavage enzyme complex, which catalyzes the rate-limiting reaction in progesterone biosynthesis, and that the VIP effect is at least partially mediated through cAMP. It is suggested that a stimulatory action of VIP on the synthesis of ovarian progesterone may contribute to regulating the functional development of the ovary.