In vitro effects of peroxisome proliferator-activated receptor-γ ligands on gene expression in lipopolysaccharide-induced endometrial and endometriotic stromal cells.

This in vitro study demonstrates the comparative anti-inflammatory effects of rosiglitazone and 15d-PGJ(2) in endometriosis and provides evidence for exploitation of peroxisome proliferator-activated receptor-γ as a therapeutic target.

Implication of the RAGE-EN-RAGE axis in endometriosis.

OBJECTIVE: To investigate the involvement of the receptor gene for advanced glycation (RAGE), its ligand EN-RAGE, and COX-2 in endometriosis. METHODS: The mRNA and protein expression of the corresponding genes were determined from endometriotic cells from 28 study patients and healthy endometrial stromal cells from 20 controls by semiquantitative RT-PCR and Western blot analysis, respectively, using beta-actin as an invariant control. RESULTS: The expression of COX-2, RAGE, and EN-RAGE was significantly increased, as evidenced by the significantly greater mRNA and protein expression in the cells of the study patients (P<0.001). Previous treatment for endometriosis did not lessen mRNA and protein expression (P<0.001). CONCLUSION: Our findings strengthen the hypothesis of an underlying inflammation in the pathophysiology of endometriosis and suggest exploring anti-inflammatory therapies as adjunct treatment.

Role of 8-iso-prostaglandin F2alpha and 25-hydroxycholesterol in the pathophysiology of endometriosis.

OBJECTIVE: To investigate the involvement of 8-iso-PGF(2alpha) and 25-hydroxycholesterol (25-OH-Chol) in the pathophysiology of endometriosis. DESIGN: Observational case-control study using enzyme immunoassay and high-performance liquid chromatography (HPLC). SETTING: Postgraduate Institute of Medical Education and Research. PATIENT(S): Forty-five women undergoing laparoscopy (n = 25), laparotomy (n = 19), or tubal ligation (n =1). INTERVENTION(S): Venipuncture and laparoscopic peritoneal fluid (PF) collection. MAIN OUTCOME MEASURE(S): The levels of 8-iso-PGF(2alpha) were determined both in urine and PF of all the patients using enzyme immunoassay. The levels of 25-OH-Chol were determined by using reversed phase HPLC both in the plasma and PF samples. Oxidative damage to DNA was assessed by agarose gel electrophoresis. RESULT(S): Significantly increased levels of 8-iso-PGF(2alpha) were observed both in urine and PF of women with endometriosis compared with control women. Similarly, higher levels of 25-OH-Chol were observed both in plasma and PF of patients compared with controls and the difference was statistically significant. A clear-cut tailing pattern was observed in DNA of patients with endometriosis, indicating significant DNA damage. CONCLUSION(S): Our observations implicate oxidative stress in the pathophysiology of endometriosis. For the first time, we demonstrate that 8-iso-PGF(2alpha) and oxysterols (the known promoters of steroidogenesis) might be the culprits in this disease.

In vitro effects of atorvastatin on lipopolysaccharide-induced gene expression in endometriotic stromal cells.

OBJECTIVE: To investigate the in vitro effects of atorvastatin on lipopolysaccharide (LPS)-induced gene expression in endometrial-endometriotic stromal cells. DESIGN: In vitro experimental study using flow cytometry, ELISA, semiquantitative reverse transcriptase polymerase chain reaction, and Western blot. SETTING: Postgraduate Institute of Medical Education and Research. PATIENT(S): Twenty-five women undergoing laparoscopy (n = 10) and laparotomy (n = 15). INTERVENTION(S): Endometriotic cyst wall (group I) and endometrial biopsy (group II) collection. MAIN OUTCOME MEASURE(S): The endometrial-endometriotic stromal cells were isolated from ectopic (group I) and eutopic (group II) endometrium by established methods, cultured, and stimulated with LPS (1 µg/mL), followed by atorvastatin treatment in a time- and dose-dependent manner to investigate the effects of LPS on proliferation (Ki-67) and expression of cyclooxygenase-2 (COX-2), vascular endothelial growth factor (VEGF), receptor for advanced glycation end products (RAGE), extracellular newly identified RAGE binding protein (EN-RAGE), peroxisome proliferator activated receptor-γ (PPAR-γ), and liver X receptor-α (LXR-α) genes in endometrial-endometriotic stromal cells and on levels of insulin-like growth factor binding protein-1 (IGFBP-1) and 17ß-E(2) in endometrial-endometriotic stromal cell culture supernatant. RESULT(S): Significant inhibition of Ki-67 and LPS-induced expression of inflammatory and angiogenic genes (COX-2, VEGF, RAGE, and EN-RAGE) was observed in atorvastatin-treated endometrial-endometriotic stromal cells. In contrast, a significant dose- and time-dependent increase in expression of anti-inflammatory genes (PPAR-γ and LXR-α) and levels of IGFBP-1 was observed after atorvastatin treatment in both the groups. However, atorvastatin treatment had no effect on 17ß-E(2) levels in endometrial/endometriotic stromal cell culture supernatant. CONCLUSION(S): The data of the present study provide new insights for the implication of atorvastatin treatment for endometriosis in humans.