Thiazolidinediones, Agonisten des PPAR-γ-Rezeptors (peroxisomaler proliferatoraktivierter Rezeptor) reduzieren die peritoneale Makrophagenanzahl - Neue medikamentöse Therapieoptionen für die Inflammation und Infertilität bei Endometriose? -

und Methode Die intraperitoneale Injektion von Thioglycollat-Medium in 7 Wochen alte, weibliche C57 BL/6-Mäuse wurde dazu benutzt, eine Rekrutierung von aktivierten Peritonealmakrophagen hervorzurufen. Die Peritonealflüssigkeit wurde gesammelt und die Peritonealzellen gezählt. Immunzytochemie wurde mit spezifischen PPAR-γ-Antikörpern durchgeführt. RNA wurde aus Peritoneal-Leukozyten isoliert und RT-PCR mit c-fms-spezifischen Primern angewendet. Ergebnisse Die intraperitoneale Injektion von Thioglycollat führte zu einer Rekrutierung von aktivierten Peritonealzellen. Die gleichzeitige, intraperitoneale Injektion von Ciglitazone reduzierte die peritoneale Leukozyteninfiltration signifikant. Spezifische, neutralisierende RANTES-Antikörper reduzierten ebenfalls die Makrophagenanzahl und dies deutet darauf hin, dass RANTES eine wesentliche Rolle bei der inflammatorischen Antwort spielt. Immunzytochemie demonstrierte, dass CD68-positive Makrophagen den PPAR-γ-Rezeptor enthalten. RT-PCR mit c-fms-spezifischen Primern bestätigte den monozytischen Phänotyp dieser Zellen. Schlussfolgerung Aktivatoren von PPAR-γ könnten für die Behandlung der endometrioseassoziierten Inflammation und Infertilität erprobt werden.

Purpose Migration of macrophages into the peritoneal cavity is an important pathophysiological feature of endometriosis. Prior data from our laboratory suggest that RANTES (Regulated on Activation, Normal T cell Expressed and Secreted) is a potential target for therapies designed to mitigate the inflammatory response associated with infertility and pain in this condition. We characterized the inflammatory response in an in vivo murine model of peritonitis and assessed the anti-inflammatory effect of ciglitazone, a thiazolidinedione (TZD) and PPAR-γ activator (peroxisome proliferator activated receptor γ).

Intraperitoneal injection of thioglycollate solution into 7-week-old female C57 BL/6 mice was used to elicit the recruitment of activated peritoneal leukocytes. Pelvic fluid was collected and peritoneal cells were counted. Immunocytochemistry with specific PPAR-γ-antibodies were afforded. RNA from peritoneal leukocytes was isolated and RT-PCR was performed with c-fms-specific primers.

Intraperitoneal injection of thioglycollate elicited the recruitment of activated peritoneal cells. Concomitant peritoneal injection of ciglitazone reduced peritoneal leukocyte infiltration significantly. Specific neutralizing anti-RANTES antibodies also reduced the number of macrophages, indicating that RANTES played a major role in the inflammatory response. Immunocytochemistry demonstrated that CD68 positive macrophages contained the PPAR-γ receptor. RT-PCR with c-fms-specific primers confirmed the monocytic phenotype of these cells.

Activators of PPAR-γ could be developed for the treatment of inflammation and infertility associated with endometriosis. Schlüsselwörter Endometriose - Peritonealmakrophagen - PPAR-γ - Thiazolidinedione Key words Endometriosis - Peritoneal macrophages - PPAR-γ - Thiazolidinedione Literatur - 1 Halme J, Becker S, Hammond M, Raj M, Raj S. Increased activation of pelvic macrophages in infertile women with mild endometriosis. Am J Obstet Gynecol. 1983; 145 333-337 - 2 Klein N A, Pergola G M, Rao-Tekmal R, Dey T D, Schenken R S. Enhanced expression of resident leukocyte interferon gamma mRNA in endometriosis. Am J Reprod Immunol. 1993; 30 74-81 - 3 Hornung D, Bentzien F, Wallwiener D, Kiesel L, Taylor R N. Chemokine bioactivity of RANTES in endometriotic and normal endometrial stromal cells and peritoneal fluid. Mol Hum Reprod. 2001; 7 163-168 - 4 Lebovic D I, Mueller M D, Taylor R N. Immunobiology of endometriosis. Fertil Steril. 2001; 75 1-10 - 5 Mueller M D, Lebovic D I, Garrett E, Taylor R N. Neutrophils infiltrating the endometrium express vascular endothelial growth factor: potential role in endometrial angiogenesis. Fertil Steril. 2000; 74 107-112 - 6 Taylor R N, Ryan I P, Moore E S, Hornung D, Shifren J L, Tseng J F. Angiogenesis and macrophage activation in endometriosis. Ann NY Acad Sci. 1997; 828 194-207 - 7 Hill J A, Anderson D J. Lymphocyte activity in the presence of peritoneal fluid from fertile women and infertile women with and without endometriosis. Am J Obstet Gynecol. 1989; 161 861-864 - 8 Hornung D, Ryan I P, Chao V A, Vigne J-L, Schriock E D, Taylor R N. Immunolocalization and regulation of the chemokine RANTES in human endometrial and endometriosis tissues and cells. J Clin Endocrinol Metab. 1997; 82 1621-1628 - 9 Hornung D, Klingel K, Dohrn K, Kandolf R, Wallwiener D, Taylor R N. RANTES mRNA expression in normal endometrium and endometriotic implants: Assessment of autocrine/paracrine regulation by in situ hybridization. Am J Pathol. 2001; 158 1949-1954 - 10 Hornung D, Waite L L, Ricke E A, Bentzien F, Wallwiener D, Taylor R N. Nuclear receptors PPAR-alpha and gamma have opposing effects on monocyte chemotaxis in endometriosis. J Clin Endocrinol Metab. 2001; 86 3108-3114 - 11 D'Hooghe T M, Pudney J, Hill J A. Immunobiology of the reproductive tract in a female baboon. Am J Primatol. 2001; 53 47-54 - 12 Rier S E, Turner W E, Martin D C, Morris R, Lucier G W, Clark G C. Serum levels of TCDD and dioxin-like chemicals in rhesus monkeys chronically exposed to dioxin: Correlation of increased serum PCB levels with endometriosis. Toxicol Sci. 2001; 59 147-159 - 13 Schenken R S, Williams R F, Hodgen G D. Experimental endometriosis in monkeys. Ann NY Acad Sci. 1991; 622 242-255 - 14 Nothnick W B, Curry T E, Vernon M W. Immunomodulation of rat endometriotic implant growth and protein production. Am J Reprod Immunol. 1994; 31 151-162 - 15 Sharpe-Timms K L, Zimmer R L, Jolliff W J, Wright J A, Nothnick W B, Curry T E. Gonadotropin-releasing hormone agonist (GnRH-a) therapy alters activity of plasminogen activators, matrix metalloproteinases, and their inhibitors in rat models for adhesion formation and endometriosis: potential GnRH-a-regulated mechanisms reducing adhesion formation. Fertil Steril. 1998; 69 916-923 - 16 Bruner K L, Eisenberg E, Gorstein F, Osteen K. Progesterone and transforming growth factor-beta coordinately regulate suppression of endometrial matrix metalloproteinases in a model of experimental endometriosis. Steroids. 1999; 64 648-653 - 17 Tabibzadeh S, Miller S, Dodson W C, Satyaswaroop P G. An experimental model for the endometriosis in athymic mice. Front Biosci. 1994; 4 C4-9 - 18 Steinleitner A, Lambert H, Lauredo I. Heterologous transplantation of activated murine peritoneal macrophages inhibits gamete interaction in vivo: a paradigm for endometriosis-associated subfertility. Fertil Steril. 1990; 54 725-729 - 19 Haney A F, Newbold C T, Doty E. An elicited intraperitoneal inflammatory response has no effect on the establishment of pregnancy in the mouse. Fertil Steril. 1994; 61 956-962 - 20 Matthei S, Stumvoll M, Kellerer M, Häring H-U. Pathophysiology and pharmacological treatment of insulin resistance. Endocrine Rev. 2000; 21 585-618 - 21 Bordji K, Grillasca J P, Gouze J N, Magdalou J, Schohn H, Keller J M, Bianchi A, Dauca M, Netter P, Terlain B. Evidence for the presence of peroxisome proliferator-activated receptor (PPAR) α and γ and retinoid Z receptor in cartilage. PPARγ activation modulates the effects of interleukin-1β on rat chondrocytes. J Biol Chem. 2000; 275 12243-12250 - 22 Su C G, Wen X, Bailey S T, Jiang W, Rangwala S M, Keilbaugh S A, Flanigan A, Murthy S, Lazar M A, Wu G D. A novel therapy for colitis utilizing PPAR-γ ligands to inhibit the epithelial inflammatory response. J Clin Invest. 1999; 104 383-389 - 23 Khorram O, Taylor R N, Ryan I P, Schall T J, Landers D V. Peritoneal fluid concentrations of the cytokine RANTES correlate with the severity of endometriosis. Am J Obstet Gynecol. 1993; 169 1545-1549 - 24 Nothnick M B. Treating endometriosis as an autoimmune disease. Fertil Steril. 2001; 76 223-231 - 25 Ho M K, Springer T A. Tissue distribution, structural characterization, and biosynthesis of Mac-3, a macrophage surface glycoprotein exhibiting molecular weight heterogeneity. J Biol Chem. 1983; 10 636-642 - 26 Jackson S M, Parhami F, Xi X P, Berliner J A, Hsueh W A, Law R E, Demer L L. Peroxisome proliferator-activated receptor activators target human endothelial cells to inhibit leukocyte-endothelial interaction. Arterioscler Thromb Vasc Biol. 1999; 19 2094-2104 - 27 Clark R B, Bishop-Bailey D, Estrada-Hernandez T, Hla T, Puddington L, Padula S J. The nuclear receptor PPAR γ and immunoregulation: PPAR γ mediates inhibition of helper T cell responses. J Immunol. 2000; 164 1364-1371 - 28 Felberbaum B, Karck U. GnRH analogues in gynecology: Agonists and antagonists. Geburtsh Frauenheilk. 1997; 57 539-544 Dr. Daniela Hornung Universitätsfrauenklinik Calwerstraße 7 72076 Tübingen Email: [email protected]