Takagi K
Maternal and Perinatal Center, Tokyo Women's Medical College, Japan.
Nihon Naibunpi Gakkai Zasshi. 1988 Oct 20;64(10):1024-37. doi: 10.1507/endocrine1927.64.10_1024.
Activities of the steroidogenic enzymes involved in functional luteolysis and PGF2 alpha induced luteolysis were determined in PMSG-hCG primed immature rats to elucidate the luteolytic effects of PGF2 alpha. Plasma progesterone (P4), plasma 20 alpha dihydroprogesterone (20 alpha OHP4), in vitro production of pregnenolone (P5) from endogenous cholesterol in ovarian mitochondria (8,000 X g pellet; Mt), activities of 3 beta hydroxysteroid dehydrogenase (3 beta HSD) in ovarian microsome (105,000 X g pellet; Ms) and 20 alpha hydroxysteroid dehydrogenase (20 alpha HSD) in ovarian cytosol(105,000 X g supernatant; Sup) were determined. For the investigation of intramitochondrial transport of cholesterol, measurement of free cholesterol (FCh) of Mt and the Lineweaver-Burk plotting for cholesterol side-chain cleavage enzyme (CSCC), prepared by osmotic shock and sonication of Mt, were carried out. Functional life span of the rat corpus luteum was estimated as 12 days post hCG treatment from plasma P4. As plasma P4 decreased, concomitant increase in plasma 20 alpha OHP4 was observed. In vitro production of P5 in Mt correlated well with plasma P4 levels, indicating cholesterol side-chain cleavage reaction is the rate limiting step involved in ovarian steroidogenesis. In contrast, both values of Km and Vmax of CSCC did not change between day 7, on which the highest value of plasma P4 observed, and day 12, on which functional luteolysis was ascertained from plasma P4. Both FCh of Mt and the activity of 3 beta HSD in Ms remained unchanged during the functional life of the corpus luteum. The activity of 20 alpha HSD in Sup increased from day 10 post hCG treatment in accordance with plasma 20 alpha OHP4. These results indicate that 1) decrease in availability of FCh to CSCC within mitochondria and 2) increase in catabolism of P4 into inactive progestin, 20 alpha OHP4, may play key roles in the functional luteolysis. To compare the events observed in functional luteolysis with PGF2 alpha induced luteolysis, the animals were treated either with PGF2 alpha (1 mg/rat; s.c.), cycloheximide (5 mg/rat; i.p.; CX), or vehicle on day 7 post hCG treatment, and sacrificed 1 hour later for the analyses mentioned above. Both plasma P4 and in vitro production of P5 in Mt decreased significantly with PGF2 alpha or CX treatment, whereas the FCh of Mt and the activity of CSCC remained unchanged with PGF2 alpha treatment. Both plasma 20 alpha OHP4 and 20 alpha HSD in Sup increased with PGF2 alpha treatment. However, CX suppressed both of them.(ABSTRACT TRUNCATED AT 400 WORDS)
为阐明前列腺素F2α(PGF2α)的溶黄体作用,我们测定了孕马血清促性腺激素(PMSG)-人绒毛膜促性腺激素(hCG)预处理的未成熟大鼠中参与功能性黄体溶解和PGF2α诱导的黄体溶解的类固醇生成酶的活性。测定了血浆孕酮(P4)、血浆20α-二氢孕酮(20α-OHP4)、卵巢线粒体(8000×g沉淀;Mt)中内源性胆固醇体外生成孕烯醇酮(P5)的量、卵巢微粒体(105000×g沉淀;Ms)中3β-羟基类固醇脱氢酶(3β-HSD)的活性以及卵巢胞质溶胶(105000×g上清液;Sup)中20α-羟基类固醇脱氢酶(20α-HSD)的活性。为研究胆固醇的线粒体内转运,对经渗透休克和超声处理的Mt进行了游离胆固醇(FCh)测定以及胆固醇侧链裂解酶(CSCC)的Lineweaver-Burk作图。根据血浆P4,将大鼠黄体的功能寿命估计为hCG处理后12天。随着血浆P4下降,观察到血浆20α-OHP4相应增加。Mt中P5的体外生成与血浆P4水平密切相关,表明胆固醇侧链裂解反应是卵巢类固醇生成中的限速步骤。相比之下,在血浆P4最高值出现的第7天和根据血浆P4确定发生功能性黄体溶解的第12天之间,CSCC的Km和Vmax值均未改变。在黄体的功能期内,Mt的FCh和Ms中3β-HSD的活性均保持不变。Sup中20α-HSD的活性从hCG处理后第10天开始增加,与血浆20α-OHP4一致。这些结果表明:1)线粒体内CSCC可利用的FCh减少;2)P4分解代谢为无活性孕激素20α-OHP4增加,可能在功能性黄体溶解中起关键作用。为比较功能性黄体溶解与PGF2α诱导的黄体溶解中观察到的事件,在hCG处理后第7天,对动物分别给予PGF2α(1mg/大鼠;皮下注射)、放线菌酮(5mg/大鼠;腹腔注射;CX)或溶剂,1小时后处死进行上述分析。PGF2α或CX处理后,血浆P4和Mt中P5的体外生成均显著降低,而PGF2α处理后Mt的FCh和CSCC的活性保持不变。PGF2α处理后,血浆20α-OHP4和Sup中20α-HSD均增加。然而,CX抑制了两者。(摘要截断于400字)