Regulation of 11beta-hydroxysteroid dehydrogenase type 1 gene expression by LH and interleukin-1beta in cultured rat granulosa cells

Abstract

Granulosa cells from preovulatory follicles show increased expression of 11 -hydroxysteroid dehydrogenase type 1 (11 HSD1) at the time of ovulation. As ovulation may be an inflammatory process, this may be a mechanism of local enhancement of the activity of anti-inflammatory glucocorticoids. In this study, we examined direct effects of LH, the proinflammatory cytokine, interleukin-1 (IL-1 ), and pharmacological activators of protein kinase A (PKA) (forskolin and dibutyryl (db) cAMP) and PKC (LHreleasing hormone and phorbol 12-myristate 13-acetate (PMA)) signalling on the expression of 11 HSD1 mRNA in vitro. Granulosa cells from immature female rat ovaries were cultured (pretreatment) in serum-free medium 199 containing recombinant human (rh) FSH (1 ng/ml) for 48 h to induce responsiveness to LH. Cell monolayers were then washed and cultured (test treatment) for a further 12 h in the presence of rhLH (0-100 ng/ml), IL-1 (0-50 ng/ml), or both. Total RNA was extracted from granulosa cell monolayers and taken for quantitative ribonuclease protection analysis of 11 HSD1 mRNA. The low level of 11 HSD1 mRNA detectable in unstimulated (control) cultures was increased approximately twofold by the 48-h pretreatment with rhFSH. Subsequent exposure to rhLH (1-100 ng/ml) for a further 12 h dose-dependently increased 11 HSD1 mRNA expression by an additional two-to threefold. Forskolin (10 µM), db-cAMP (2 mM), LH-releasing hormone (LHRH; 1 µM) and PMA (200 nM) were also stimulatory. IL-1 (0·05-50 ng/ml) stimulated 11 HSD1 mRNA expression in a dose-related manner, both in the absence and in the presence of rhLH (3 ng/ml). The interaction between IL-1 (5 ng/ml) and rhLH (3 ng/ml) was additive. Cotreatment with a 50-fold excess of IL-1 receptor antagonist fully reversed the action of IL-1 . We conclude that 11 HSD1 mRNA expression in functionally mature granulosa cells is directly stimulated by gonadotrophins and IL-1 in vitro, potentially involving post-receptor signalling via PKA-and PKC-mediated pathways. Thus both LH and IL-1 may serve physiological roles in the upregulation of 11 HSD1 gene expression by granulosa cells in ovulatory follicles.

Gene Expression of 11.BETA.-HSD and Glucocorticoid Receptor in the Bovine (Bos taurus) Follicle During Follicular Maturation and Atresia: The Role of Follicular Stimulating Hormone

“…FSH appears to be responsible for the upregulation of 11HSD1 gene expression in bovine GCs, as has been shown in cultured human and rat GCs [12,35]. The expression of 11HSD2, on the other hand, was not affected by FSH, implying that the alteration of glucocorticoids metabolism associated with follicular maturation is primarily regulated by FSH-responsive 11HSD1 in bovine GCs.…”

Glucocorticoid metabolism in the bovine cumulus–oocyte complex matured in vitro

“…This is in accordance with the previous findings where FSH has been shown to increase the expression of HSD11B1 in the cultured granulosa cells of rats (Tetsuka et al 1999a) and cattle (Tetsuka et al 2010).…”

“…Two glucocorticoid metabolizing enzymes, 11b-hydroxysteroid dehydrogenase type 1 (HSD11B1) and type 2 (HSD11B2), are responsible for modulating the levels of active glucocorticoid in target organs: HSD11B1 is a bidirectional HSD11B that mainly activates (reduces) cortisone to cortisol (11-dehydrocorticosterone to corticosterone in rodents), whereas HSD11B2 is a dehydrogenase that inactivates (oxidize) cortisol to cortisone (Albiston et al 1994, Michael et al 2003. These HSD11Bs are expressed in various ovarian tissues and their expressions are regulated by gonadotropins and paracrine/autocrine factors (Tetsuka et al 1999a, Michael et al 2003. The expression of HSD11B1 is dramatically increased, while that of HSD11B2 decreased after the preovulatory gonadotropin surge or LH/hCG treatment in human (Tetsuka et al 1997), macaque (Fru et al 2006) and rat (Tetsuka et al 1999b) follicles/ovaries as well as cultured granulosa cells of rats (Tetsuka et al 1999a) and macaques (Fru et al 2006).…”

“…These HSD11Bs are expressed in various ovarian tissues and their expressions are regulated by gonadotropins and paracrine/autocrine factors (Tetsuka et al 1999a, Michael et al 2003. The expression of HSD11B1 is dramatically increased, while that of HSD11B2 decreased after the preovulatory gonadotropin surge or LH/hCG treatment in human (Tetsuka et al 1997), macaque (Fru et al 2006) and rat (Tetsuka et al 1999b) follicles/ovaries as well as cultured granulosa cells of rats (Tetsuka et al 1999a) and macaques (Fru et al 2006). Likewise, both dehydrogenase and reductase activities brought about by HSD11B1, but not by HSD11B2, have been demonstrated in cultured human granulosa-lutein cells collected shortly before ovulation (Thomas et al 1998).…”

Anti-inflammatory steroid signalling in the human peritoneum

“…From work on renal mesangeal cells it emerged that inflammatory cytokines such as interleukin-1a (IL-1a) and tumour necrosis factor a directly stimulate 11bHSD1 and suppress 11bHSD2 activities in vitro (Escher et al 1997). This is now known to hold for several other cell types that naturally respond to inflammatory signals, including ovarian granulosa (Tetsuka et al 1999) and surface epithelial cells (Yong et al 2002), bronchial epithelial cells (Feinstein & Schleimer 1999), preadipocytes (Tomlinson et al 2001), osteoblasts , aortic smooth muscle cells (Cai et al 2001), peritoneal macrophages (Gilmour et al 2006), trophoblast (Li et al 2006) and fetal membranes (Sun & Myatt 2003).…”