title: Insulin enhances FSH-stimulated steroidogenesis by cultured rat granulosa cells.
abstract: Experimentally induced diabetes in female rats is associated with decreased ovarian functions. We have investigated the ovarian action of insulin using granulosa cells obtained from immature hypophysectomized, estrogen-treated rats. The cells were cultured for 2 days in a serum-free medium in the presence of follicle-stimulating hormone (FSH), with or without insulin. Medium steroids were determined by specific radioimmunoassay. Treatment with FSH caused a dose-dependent increase in the production of estrogen, progesterone, 20 alpha-hydroxypregn-4-en-3-one (20 alpha-OH-progesterone) and pregnenolone. Concomitant treatment with 100 ng/ml insulin increased the responsiveness of granulosa cells to FSH, decreasing the ED50 values for FSH-stimulated estrogen and progestin production 2-2.75-fold. A lower dose of insulin, 10 ng/ml, also augmented FSH action. In contrast, treatment with insulin alone had no effect on steroid production. The insulin effect on progestin and estrogen biosynthesis was detected by 24 and 44 h after treatment, respectively. Also, the insulin action appeared to be specific since an insulin fragment, desoctapeptide insulin, exhibited no effect. Insulin also increased the FSH-stimulated increase in activity of 3 beta-hydroxysteroid dehydrogenase, the rate-limiting enzyme in the formation of progesterone, but was without effect on the activity of 20 alpha-hydroxysteroid dehydrogenase, which converts progesterone to the inactive 20 alpha-OH-progesterone. The effects of insulin on increasing FSH responsiveness could not be accounted for by changes in cell viability or total cell number. These results indicate that insulin exerts a specific action on granulosa cells to increase the FSH stimulation of estrogen and progestin production.(ABSTRACT TRUNCATED AT 250 WORDS)
mesh_terms: D015089:20-Hydroxysteroid Dehydrogenases / Q000378:metabolism; D015096:3-Hydroxysteroid Dehydrogenases / Q000378:metabolism; D000818:Animals; D002478:Cells, Cultured; D004967:Estrogens / Q000096:biosynthesis; D005260:Female; D005640:Follicle Stimulating Hormone / Q000494:pharmacology*; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D007328:Insulin / Q000494:pharmacology*; D011372:Progestins / Q000096:biosynthesis; D051381:Rats; D011919:Rats, Inbred Strains; D013256:Steroids / Q000096:biosynthesis*; D013997:Time Factors
chemical_list: D004967:Estrogens; D007328:Insulin; D011372:Progestins; D013256:Steroids; D005640:Follicle Stimulating Hormone; D015096:3-Hydroxysteroid Dehydrogenases; D015089:20-Hydroxysteroid Dehydrogenases