Anti-inflammatory steroid signalling in the human peritoneum

Abstract

Peritoneal surface epithelial (PSE) cells participate in adhesion formation following inflammatory injury yet adjacent ovarian SE (OSE) cells regenerate without scarification after ovulation. OSE cells show inflammation-associated expression of 11b hydroxysteroid dehydrogenase type 1 (11bHSD1) enzyme, enabling intracrine generation of anti-inflammatory cortisol to minimise tissue damage. We asked if human PSE cells show an 11bHSD1 response to pro-/anti-inflammatory stimulation and if so, how the 11-oxoreductase activity generated compares with OSE. PSE collected from premenopausal women undergoing surgery for benign gynaecological conditions were used to establish primary PSE cell cultures that were treated for 48 h with interleukin-1a (IL-1a) with/without anti-inflammatory steroid (cortisol or progesterone). mRNA levels corresponding to the genes of interest (11bHSD1, 11bHSD2, cyclooxygenase-2, COX-2) were measured by quantitative RT-PCR. IL-1a (0 . 5 ng/ml) stimulated 11bHSD1 and COX-2 mRNA levels in PSE cells but 11bHSD2 was unaffected. Cortisol (1 mM), not progesterone (1 mM), increased 11bHSD1 mRNA and synergistically enhanced IL-1a action. Cortisol suppressed IL-1a-stimulated COX-2 more effectively than progesterone. PSE cells had a significantly lower basal 11-oxoreductase enzyme activity than OSE cells; IL-1a did not significantly increase the 11-oxoreductase activity in PSE cells but did so in OSE cells. We conclude that PSE cells respond to IL-1a and antiinflammatory steroids in qualitatively similar ways as OSE. However, the enzymatic activity of 11bHSD1 is lower in PSE and less responsive to IL-1a. This could help explain why peritoneal healing often leads to adhesion formation, whereas postovulatory ovarian healing is scar-free.

Dynamics of the primate ovarian surface epithelium during the ovulatory menstrual cycle

“…The importance of inflammation is also supported, by the expression of glucocorticoid receptors and metabolic enzymes, not reported here, that may mediate proliferation, attenuate local inflammatory damage and protect the genome [e.g. 11bHSD1 and 2, and IL-1R; (Syed et al, 2001;Gubbay et al, 2004;Rae et al, 2004;Fegan et al, 2008) and unpublished microarray and IHC observations].…”

“…The molecular basis for why the OSE is so highly prone to transformation is not understood, but because the major risk factors for the disease are age, menopause, ovarian function and genetics (Sueblinvong and Carney, 2009), it would appear that a woman's natural biology places these cells at risk. A considerable body of data from human tissues, cell culture experiments and animal studies provides rationale for how these factors may promote EOC: (i) the frequency of premalignant inclusion cysts correlates positively with age and the total number of ovulations a woman experiences (Westhoff et al, 1993;Heller et al, 2005;Tan et al, 2005); (ii) progestins can induce OSE apoptosis in vivo (Rodriguez et al, 2002); (iii) cultured OSE and ovarian cancer cells respond to high concentrations of estradiol and progesterone (E and P) by growth arrest and up-regulation of apoptotic indicators (Keith Bechtel and Bonavida, 2001;Wright et al, 2005), or by increased proliferation in vivo (Bai et al, 2000); (iv) gonadotrophins that regulate ovarian function can promote proliferation in cultured OSE cells and in vivo (Davies et al, 1999;Ivarsson et al, 2001;Parrott et al, 2001); (v) ovulation is accompanied by wound/inflammatory processes that are genotoxic (Gubbay et al, 2005;Rae and Hillier, 2005;Fegan et al, 2008); (vi) prior to ovulation, ovarian factors inflict genetic damage and death on the OSE (Colgin and Murdoch, 1997;Murdoch et al, 2001;Murdoch and Martinchick, 2004), whereas after ovulation, increased proliferation in the OSE has been detected (Osterholzer et al, 1985;Bai et al, 2000); and (vii) models of superovulation show increased OSE proliferation and inclusion cyst formation (Burdette et al, 2006(Burdette et al, , 2007.…”

The Peritoneum Is Both a Source and Target of TGF-β in Women with Endometriosis

“…Peritoneal fluid (5–10 ml) was collected from women with (n = 6) and without (n = 6) endometriosis and stored in cryovials at −80°C for later analysis. HPMCs (n = 3) were isolated at the time of surgery as previously described by gentle brushing the pelvic mesothelium with a Tao™ brush followed by vigorously agitating in 15 ml of serum-containing culture media to dislodge cells before transferring to a 75 cm 2 culture flask and incubated at 37°C under 5% CO 2 in air (QC Sciences, Virginia, USA) [25].…”

“…Brushings of HPMC were cultured in HOSE1 media containing; 40% media 199, 40% MCDB 105 and supplemented with 15% FBS, 0.5% penicillin/streptomycin and 1% L-glutamine, as previously described [25] (Life Technologies Inc., Paisley UK and Sigma Chemical Co., Poole UK). For immunoassay, cells were cultured in serum free conditions for between 12 and 48 hrs.…”

Prostaglandin F2alpha Stimulates 11Beta-Hydroxysteroid Dehydrogenase 1 Enzyme Bioactivity and Protein Expression in Bovine Endometrial Stromal Cells1

“…The inhibitory effect of cortisol on PGF2a production in bovine endometrial stromal cells [7], as well as the stimulatory effect of GC on intrauterine PGs in pregnant ewes [14] and human placental membrane [15,16], suggests the presence of a dynamic interaction between PGs and GC in the uterus depending on the reproductive status. Although PGF2a has the ability to increase cortisol production by stimulating HSD11B1 in human ovarian surface epithelial cells [17] and in chorion trophoblast [12,18], the mechanisms by which PGF2a increases the biological action of HSD11B1 in bovine endometrium are poorly understood.…”