Biomedical subjects

W B Nothnick

Publications and source records attributed to W B Nothnick.

18 recordsLinked to original sources

PubMed2001

Treating endometriosis as an autoimmune disease.

OBJECTIVE: To review the literature on the role of autoimmunity in the etiology of endometriosis, compare the similarities in the pathophysiologies between endometriosis and autoimmune diseases, and discuss the use of immunomodulators currently used to treat autoimmune diseases as potential therapies for endometriosis. DESIGN: The literature on endometriosis and other autoimmune diseases was reviewed, and summary data are presented. RESULTS: Endometriosis shares many similarities with autoimmune diseases such as rheumatoid arthritis, Crohn's disease, and psoriasis. These similarities include elevated levels of cytokines, decreased cell apoptosis, and T- and B-cell abnormalities. Because the use of immunomodulators and inflammatory modulators has proven to be an effective means of medical management for these autoimmune diseases, similar therapies may prove useful in treating endometriosis. CONCLUSION(S): Although substantial evidence indicates that endometriosis at least shares many similarities with autoimmune diseases, endometriosis is primarily treated by using compounds that induce a hypoestrogenic environment. A review of the literature combined with the shortcomings of current means of medical management for endometriosis support the postulate that treatment of endometriosis with immunomodulators and inflammatory modulators is warranted.

Adjuvants, Immunologic↗

PubMed2001

Disruption of the tissue inhibitor of metalloproteinase-1 gene in reproductive-age female mice is associated with estrous cycle stage-specific increases in stromelysin messenger RNA expression and activity.

Tissue inhibitors of metalloproteinases (TIMPs) are expressed in the uteri of virtually all species, yet the precise role of these factors in uterine physiology is uncertain. It has been previously demonstrated that disruption of the TIMP-1 gene product in vivo results in altered reproductive cycles and an aberrant uterine phenotype. Because this phenotype may be due to an elevation in uterine matrix metalloproteinase (MMP) activity, the purpose of the following experiments was to identify which uterine MMPs may have their expression altered in response to disruption of the TIMP-1 gene. Mature female TIMP-1 wild-type and null mice were killed during each stage of the estrous cycle, and uterine MMP activity and transcript expression were assessed. Disruption of the TIMP-1 gene product was associated with an increase in total uterine protease activity. Gel zymography further revealed that uterine stromelysin (stromelysin-1, -2, and -3) activity was significantly increased in the TIMP-1 null mice, whereas Northern blot analysis indicated that an up-regulation of stromelysin-1 and -3 mRNA expression may contribute to this increase in activity. It is concluded from this study that TIMP-1 plays a pivotal role in regulating uterine stromelysins both at the level of protease activity and the level of transcript expression.

Animals↗

PubMed2001

CD44s expression is reduced in endometriotic lesions compared to eutopic endometrium in women with endometriosis.

Immunohistochemical expression of the standard form of CD44 (CD44s) was examined in archival formalin-fixed endometriotic and matching eutopic endometrial tissue obtained from 25 patients in proliferative (N = 16) and secretory (N = 9) stages of the cycle. CD44s was expressed in most eutopic endometria and endometriotic tissue. Its expression was significantly higher in secretory than in proliferative phase endometrium. It was low but detectable in 13 of 16 proliferative phase biopsies. The majority of these endometria exhibited both glandular and stromal staining (63%). In the secretory phase, glandular cells exhibited a significantly greater intensity of staining compared to stromal cells. In endometriotic tissue, stromal cell CD44s expression did not differ between tissue types in either stage of the cycle. In contrast, glandular expression in endometriotic tissue during the secretory phase was reduced (p < 0.05) compared to eutopic endometrium. It was absent in 66% of cases and reduced in the remaining cases. Our results indicate a correlation between CD44s expression and secretory differentiation of endometrial glands in the cycle, suggesting hormonal regulation of its expression. This cyclic pattern of CD44s expression was lost in corresponding endometriotic tissue. Reduced expression of CD44s in endometriotic tissue may provide insight into the pathophysiology of endometriosis.

Adult↗

PubMed2001

Reduction in reproductive lifespan of tissue inhibitor of metalloproteinase 1 (TIMP-1)-deficient female mice.

Tissue inhibitor of metalloproteinase 1 (TIMP-1) is a multifunctional protein expressed in the ovary and uterus of females of several species including mice, rats and humans. Mice deficient in TIMP-1 protein display altered reproductive parameters, which include reduced serum progesterone concentrations and abnormal uterine morphology. As these abnormalities could lead to altered reproductive performance, the objective of this study was to examine whether TIMP-1 deficiency is associated with a reduction in the reproductive lifespan of female mice in which the TIMP-1 gene is disrupted. Wild-type (n = 50) and TIMP-1 null mice (n = 90) were mated with males of proven fertility from the same genotype for 12 months during which the number of litters delivered and the number of pups per litter were determined. Significantly fewer TIMP-1 null females achieved pregnancy (47 of 90 or 52%) compared with the wild-type mice (39 of 50 or 78%; P < 0.05). TIMP-1-deficient female mice that achieved and maintained pregnancy had significantly fewer litters during the 12 months (2.9 +/- 0.8 versus 3.5 +/- 0.8; P < 0.01) and significantly fewer pups per litter (5.7 +/- 1.3 versus 4.7 +/- 1.1; P < 0.05) than did wild-type mice. Females of both genotypes produced two consecutive litters, after which significantly fewer TIMP-1 null mice became pregnant. Mating of females with males of proven fertility from the other genotype confirmed that these abnormalities were not due to the inability of TIMP-1 null males to produce offspring. These data indicate that the absence of TIMP-1 is associated with a reduction in the reproductive lifespan of female mice, which may be manifested at the ovary, uterus or both organs.

Animals↗

PubMed2000

Disruption of the tissue inhibitor of metalloproteinase-1 gene results in altered reproductive cyclicity and uterine morphology in reproductive-age female mice.

Tissue inhibitor of metalloproteinase-1 (TIMP-1) is a multifunctional protein expressed in the uterus of essentially all species, yet the function of this protein is uncertain. To assess the role of TIMP-1 in the uterine events that occur during the murine estrous cycle, mature female TIMP-1 wild-type and null mice were monitored for reproductive cyclicity. Mice were sacrificed in each stage of the estrous cycle, and peripheral blood was collected and assayed for serum estradiol and progesterone content by RIA. Uterine morphology and TIMP-1, TIMP-2, TIMP-3, and TIMP-4 mRNA expression were also examined between genotypes in each stage of the estrous cycle. Disruption of the TIMP-1 gene product was associated with an altered reproductive cycle characterized by a significant decrease in the length of the estrus period in the null mice. Also during the period of estrus, null mice expressed significantly lower levels of uterine TIMP-3 mRNA expression, altered uterine morphology, significantly higher serum estradiol levels, and significantly lower serum progesterone levels compared to their wild-type counterparts. It is concluded from this study that TIMP-1 has a multifaceted role in regulating the murine reproductive cycle, and this control appears to be at the level of both the uterus and the ovary.

Animals↗

PubMed1999

Collagenase and gelatinase messenger ribonucleic acid expression and activity during follicular development in the rat ovary.

Metalloproteinases are members of a family of proteinases that remodel the extracellular matrix throughout the body. To test the hypothesis that metalloproteinases are regulated by gonadotropin-induced changes during follicular growth, rats were injected with eCG (20 IU, s.c.), and ovaries and serum were collected at the time of eCG administration (0 h) and at 6, 12, 24, 36, or 48 h later for analysis of metalloproteinase mRNA expression, metalloproteinase activity, and steroidogenesis. Serum estradiol levels increased from 18.9 pg/ml at 0 h to 503.8 pg/ml at 48 h. Analysis of mRNA expression was performed for collagenase-3, 72-kDa gelatinase, and 92-kDa gelatinase (n = 3-4). For collagenase-3, eCG stimulated a 32-fold increase in collagenase-3 mRNA at 48 h after eCG injection as compared to that in ovaries collected at the time of eCG administration (i.e., 0-h control). The mRNA levels for 72-kDa gelatinase were 2.8-fold compared to 0 h at 36 h after eCG treatment and returned to control levels by 48 h after gonadotropin treatment. Levels of the 92-kDa mRNA expression peaked at 24 h (4. 2-fold compared to 0 h) and returned to control levels by 36 h. Gel zymography revealed 3 gelatinolytic bands corresponding to the gelatinases of approximately 72 kDa, 92 kDa, and 105 kDa. Analysis of metalloproteinase activity as the degradation of collagen or gelatin per ovary showed an increase in gelatinolytic and collagenolytic activity between 12 and 48 h after eCG treatment. In summary, these findings demonstrate that the gonadotropin induction of folliculogenesis results in changes in the metalloproteinases that may be responsible for extracellular matrix remodeling associated with follicular growth.

Animals↗

PubMed1998

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.

OBJECTIVE: To evaluate the effects of a gonadotropin-releasing hormone agonist (GnRH-a) on plasminogen activator (PA), matrix metalloproteinase (MMP), plasminogen activator inhibitor (PAI) and matrix metalloproteinase inhibitor (MMPI) activities in peritoneal fluid relative to GnRH-a-induced reduction of adhesion formation. DESIGN: Continuation of prospective randomized study using surgical models for adhesion formation. SETTING: Department of Obstetrics and Gynecology research laboratory at the University of Missouri School of Medicine. PATIENT(S): Forty reproductively cycling female Sprague-Dawley rats. INTERVENTION(S): Female rats were injected with depot GnRH-a or diluent and randomly assigned to adhesion and endometriosis surgeries. Peritoneal fluid was collected prior to (time 1) and 7 weeks from (time 2) initial surgery. MAIN OUTCOME MEASURE(S): Peritoneal fluid was analyzed for PA, PAI, MMP, and MMPI activities. RESULT(S): At time 1, MMP and MMPI activities were similar in all rats; however, PA and PAI activities were less in rats pretreated with GnRH-a than with diluent. Between time 1 and time 2, GnRH-a-treated rats showed an increase in PAI and MMPI activities without significant changes in PA or MMP activities, whereas rats receiving diluent showed a significant increase in PAI and MMP activities but no significant changes in PA or MMPI activities. At time 2, rats receiving GnRH-a had less PA and MMP activities than those receiving diluent. Adhesion scores showed a positive correlation with MMP activity. CONCLUSION(S): In the absence of GnRH-a therapy, surgical tissue manipulation increased peritoneal fluid MMP and PAI activity. Gonadotropin-releasing hormone agonist therapy decreased PA and MMP activities and also increased PAI and MMPI activities. This GnRH-a-induced shift to a less invasive phenotype may alter fibrinolysis and extracellular matrix remodeling and thereby play a role in the mechanism of GnRH-a-induced reduction in adhesion formation.

Animals↗

PubMed1998

Pattern of messenger ribonucleic acid expression of tissue inhibitors of metalloproteinases (TIMPs) during testicular maturation in male mice lacking a functional TIMP-1 gene.

It has been proposed that proteolytic remodeling of the testicular extracellular matrix (ECM) plays a fundamental role in testicular development, morphogenesis, and spermatogenesis. Tissue inhibitor of metalloproteinase (TIMP)-1 regulates ECM turnover and has been reported to stimulate Leydig cell steroidogenesis. To assess the developmental changes in TIMP mRNA expression and the potential steroidogenic role of TIMP-1 in testicular physiology, an experiment was conducted that used male mice incapable of expressing the TIMP-1 gene product. TIMP-1-deficient and wild-type male mice (n = 6 to 15 per age group per genotype) were killed at 18, 21, 24, 27, 33, 41, and 49 days of age. Body weight, testis weight, serum total testosterone, and TIMP-1, -2, -3, and -4 transcript expression were determined. Northern analysis revealed the detection of TIMP-1 mRNA in wild-type males only. TIMP-1 mRNA levels (per 20 microg total RNA) were highest in 18- to 27-day-old male mice and decreased approximately 13-fold by Day 41. The pattern of TIMP-2 expression was similar between genotypes, with testicular levels of the 1. 0-kilobase transcript increasing between Days 18 and 27 of age. The pattern of TIMP-3 transcript expression (per 20 microg total RNA) was similar between genotypes and decreased between Days 18 and 41 of age. When TIMP-3 mRNA levels were expressed on a per testis basis, TIMP-3 was seen to have increased throughout testicular development. TIMP-4 mRNA expression was undetectable by Northern analysis in all mice. No significant difference was detected in body weight or testis weight between genotypes, with the exceptions that 21-day-old TIMP-1 mutants had higher (p < 0.05) testis weights and lower (p < 0. 05) serum total testosterone levels than age-matched wild-type males. It is concluded that each TIMP displays its own unique pattern of expression during the prepubertal period, suggesting that the various TIMPs may have specific roles in testicular development. The modest effect of TIMP-1 ablation on testosterone is interpreted to mean that TIMP-1 may function as a coregulator of basal testicular steroidogenesis; but overall, TIMP-1 appears to have little effect on testosterone production in mice lacking the TIMP-1 gene.

Aging↗

PubMed1998

Novel implications in the development of endometriosis: biphasic effect of macrophage activation on peritoneal tissue expression of tissue inhibitor of metalloproteinase-1.

PROBLEM: Elevated levels of activated macrophages are associated with endometriosis, but their role in the etiology of the disease is uncertain. The current study was undertaken to examine whether activated macrophages could modulate peritoneal tissue expression of the tissue inhibitor of metalloproteinase-1 (TIMP-1), which may play a role in the development of endometriosis. METHOD OF STUDY: Female mice were treated with the macrophage activator lipopolysaccharide (LPS), and peritoneum TIMP-1 mRNA was examined by Northern blot analysis. RESULTS: LPS induced a dose-dependent increase (P < 0.05) in TIMP-1 mRNA expression at levels of 1 microgram (70.0% +/- 5.8% greater than the control), 10 micrograms (83.0% +/- 12.0% greater than the control), and 25 micrograms (100.0% +/- 10.0% greater than the control). In contrast, the administration of 50 micrograms of LPS resulted in a decrease in TIMP-1 mRNA expression below baseline levels (18.0% +/- 6.0% less than the control values). CONCLUSIONS: Activated macrophages and/or their products modulate peritoneum TIMP-1 expression. These data suggest that, in addition to their phagocytotic role in the peritoneal cavity, these immune cells also may play a novel role in influencing the ability of the peritoneum to regulate tissue/cell invasion and in the development of endometriosis through TIMP-1 expression.

Animals↗

PubMed1997

Role of direct contact between spermatozoa and oviductal epithelial cells in maintaining rabbit sperm viability.

In many species, including the rabbit, spermatozoa attach to the apical plasma membrane of oviductal epithelial cells. Attachment to oviductal epithelial cells has a beneficial effect on the maintenance of sperm viability during storage in the oviduct, but the mechanism by which this occurs is unknown. The present study was conducted to determine the role of direct contact between spermatozoa and the apical plasma membrane of oviductal cells in maintaining sperm viability. To accomplish this, an apical plasma membrane fraction was isolated from homogenized rabbit oviducts and kidney (control) by differential precipitation. When viewed by electron microscopy, this fraction was composed of closed, roughly spherical apical membrane vesicles (AMV) 50-300 nm in diameter. Analysis of anovulatory, preovulatory, and periovulatory oviductal AMV by one-dimensional PAGE revealed 19 major bands. Densitometry revealed quantitative changes in these bands in relation to reproductive stage. Rabbit spermatozoa were incubated for 48 h in the presence of anovulatory, preovulatory, and periovulatory oviductal AMV. As controls, spermatozoa were incubated with rabbit kidney AMV or in defined medium (DM) alone. There was no significant (p < 0.05) decline in viability for spermatozoa incubated with periovulatory oviductal AMV during the 48-h incubation period. In contrast, viability dropped significantly by 12 h for spermatozoa incubated with preovulatory and anovulatory oviductal AMV, kidney AMV, or DM alone. Periovulatory oviductal AMV were significantly more effective at maintaining sperm viability than preovulatory or anovulatory oviductal AMV. Sperm viability in the presence of kidney AMV was not significantly different from that in DM alone. From these data it can be concluded that direct membrane contact between spermatozoa and oviductal epithelial cells plays a role in maintaining sperm viability. Furthermore, this effect appears to be tissue specific and related to reproductive stage.

Animals↗

PubMed1997

Assessment of the role of tissue inhibitor of metalloproteinase-1 (TIMP-1) during the periovulatory period in female mice lacking a functional TIMP-1 gene.

Tissue inhibitor of metalloproteinase (TIMP)-1 is a multifunctional peptide that has been implicated in the ovulatory process. To assess the function of TIMP-1 during the periovulatory period in vivo, mice incapable of expressing the TIMP-1 gene product were utilized. Twenty-three-day-old TIMP-1-deficient (n = 59) and wild-type (n = 61) female mice were injected with 5 IU eCG, followed 48 h later by an ovulation-inducing dose of hCG (5 IU). Animals were killed at the time of hCG injection (0-h hCG), at 12 h (12-h hCG), or at 24 h post-hCG (24-h hCG) administration. Serum was collected for the assessment of estradiol-17beta (0-h hCG groups) or progesterone content (12- and 24-h hCG groups), while ovaries were removed for either histological preparation or Northern analysis of TIMP-1, TIMP-2, and TIMP-3. The number of healthy and atretic follicles was determined in the 0-h hCG groups, as was the number of oocytes released in the 24-h hCG group. TIMP-1-deficient females in the 0-h hCG group showed reduced levels of ovarian TIMP-2 (0.29-fold decrease, p < 0.05) and TIMP-3 (3.0-fold decrease, p < 0.05) expression compared to wild-type counterparts. No significant difference was detected between genotypes in the 0-h hCG group for number of healthy or atretic follicles or for serum estradiol-17beta concentrations. Additionally, no significant differences were detected between genotypes in the 12- and 24-h hCG groups for serum progesterone concentrations, ovarian TIMP-2 and TIMP-3 expression, or number of oocytes released (24-h hCG group). To assess the effect of TIMP-1 on steroidogenesis in vitro, granulosa cells were obtained from 23-day-old, eCG-primed TIMP-1-deficient and wild-type females. Addition of recombinant human TIMP-1 significantly increased conditioned media estradiol-17beta concentrations in cell cultures from both mutant (1.32-fold over controls; p = 0.02; n = 4) and wild-type females (1.16-fold over controls; p = 0.04; n = 3). It is concluded from this study that TIMP-1 may modulate ovarian TIMP-2 and TIMP-3 mRNA expression during folliculogenesis. In addition, TIMP-1 exhibits steroidogenic activity in vitro, but no evidence was found for regulation of steroidogenesis in vivo.

Animals↗

PubMed1996

Collagenase, gelatinase, and proteoglycanase messenger ribonucleic acid expression and activity during luteal development, maintenance, and regression in the pseudopregnant rat ovary.

In the present study, we examined the roles of the matrix metalloproteinases collagenase, 72-kDa and 92-kDa gelatinase, and proteoglycanase in the tissue remodeling that occurs during luteal development and regression, using a pseudopregnant rat model. Pseudopregnancy was induced in immature female rats by eCG/hCG priming, and animals (n = 3 to 4 per time point) were killed on Days 1, 2, 4, 8, 12, 14, or 16 of pseudopregnancy (Day 0 = time of hCG administration). Ovaries were then removed and analyzed for either matrix metalloproteinase mRNA expression or activity. During luteal development (Day 1 of pseudopregnancy), activity of both collagenase (p = 0.009) and gelatinase (p = 0.0003), but not proteoglycanase (p > or = 0.05), was significantly greater than at all other time points. In accord with gelatinase activity, transcript levels of this enzyme were elevated at Day 1 of pseudopregnancy. Specifically, gelatinase transcript levels for the 72-kDa and 92-kDa enzymes were greatest at Day 1 (p = 0.0003 and p = 0.001, respectively), decreased 3-fold by Day 2,4-fold by Day 4, and reached 10-fold lower levels by Days 8 and 12 of pseudopregnancy. Proteoglycanase transcript could not be detected by Northern analysis during luteal development or any other time point examined in the current study. During the period of luteal maintenance (approximately Days 4-8 of pseudopregnancy in the rat), collagenase and gelatinase displayed basal levels of activity, but only proteoglycanase activity was elevated compared to luteal development levels of this enzyme. During luteal regression (Days 12-14 of pseudopregnancy in the rat), all enzymes displayed basal levels of enzyme activity. In accord with gelatinolytic activity during luteal regression, both 72-kDa and 92-kDa gelatinase mRNA were detectable at baseline levels. In contrast to the baseline levels of collagenolytic activity during luteal regression, collagenase transcript displayed peak values (approximately 8-fold greater than Day 1 levels; p = 0.004) at Day 12 of pseudopregnancy. It is concluded from these studies that collagenase and the gelatinases play a role in the tissue remodeling associated with luteal development, proteoglycanase is associated with luteal maintenance, and collagenase may contribute to the structural regression of the corpus luteum.

Animals↗

PubMed1996

Mifepristone and ovarian function.

In summary, RU 486 has been a powerful instrument in delineating progesterone action on the ovary. However, early experiments using RU 486 must be interpreted with the understanding that systemic administration of the antiprogestin may have had extraovarian sites of action, such as at the hypothalamic-pituitary axis or at the adrenal, that in turn led to indirect ovarian responses. Treatment with progesterone, agonist, or antagonist at periods during which the ovary lacks progesterone receptors would further suggest extraovarian sites of action or nongenomic mechanisms of action. Furthermore, the dose of ligand or antagonist administered and the hormonal milieu at the time of administration may dictate the ovarian response (Espey L, personal communication). For example, low doses of exogenous progesterone may elicit a biologic response, whereas high doses are without effect or may inhibit the biologic effect observed at lower doses. Although RU 486 is classically described as an antiprogestin, agonist actions have been observed in addition to its the well documented antiglucocorticoid effects. All of these variables may contribute to the confounding observations of progesterone and RU 486 action on the ovary. Regardless of these caveats, experimental paradigms have demonstrated that RU 486, either indirectly or directly, regulates ovarian folliculogenesis, stimulates and/or inhibits steroidogenesis depending on the species and time of RU 486 administration, inhibits ovulation, and modulates luteal function. These findings supports a progesterone-dependent mechanism in these varied aspects of ovarian function.

Animals↗

PubMed1996

Divergent effects of interleukin-1 beta on steroidogenesis and matrix metalloproteinase inhibitor expression and activity in cultured rat granulosa cells.

The periovulatory increase in ovarian matrix metalloproteinase inhibitor (MMPI) expression is regulated both in vitro and in vivo by LH, but the intermediary steps in this process are uncertain. The purpose of this experiment was to determine whether interleukin-1 beta (IL-1 beta), a known modulator of MMPI expression in other systems and one that is induced by LH in the ovary, is capable of regulating granulosa matrix metalloproteinase inhibitor expression and activity. Using an in vitro rat granulosa cell model, these parameters were assessed in response to IL-1 beta or LH administration alone or in combination. Granulosa cells were obtained from 24-day-old immature female rats primed with 20 IU PMSG at 22 days of age. Cells were cultured under serum-free conditions for 24 h at 37 C in the presence of medium (control), LH (100 ng/ml), IL-1 beta (10 ng/ml), or LH plus IL-1 beta. MMPI activity in the conditioned medium was assessed using a colorimetric assay (n = 6), whereas progesterone and estrogen concentrations in the conditioned medium were determined by RIA (n = 6). RNA was isolated from the granulosa cells and assessed for Northern analysis of tissue inhibitor of metalloproteinase-1 (TIMP-1; n = 4), TIMP-2 (n = 3), and TIMP-3 (n = 3) expression. When added to granulosa cells, IL-1 beta and LH each significantly (P < 0.05) increased MMPI activity in granulosa cell-conditioned medium above control values (40.9 +/- 3.0% inhibition for IL-1 beta and 67.1 +/- 5.6% inhibition for LH vs. 31.4 +/- 2.4% inhibition for controls). When added in combination, IL-1 beta had no effect on LH-stimulated inhibitor activity (67.1 +/- 5.6% inhibition vs. 69.9 +/- 5.1% inhibition for LH and LH plus IL-1 beta, respectively). Methylamine hydrochloride treatment revealed that the majority of inhibitor activity in all treatment groups was derived from TIMPs. The patterns of TIMP-1, TIMP-2, and TIMP-3 messenger RNA expression among the treatment groups paralleled the TIMP-derived inhibitor activity, in that both IL-1 beta and LH alone stimulated transcript expression of all three TIMPs. In addition, an increase in progesterone production was associated with IL-1 beta-stimulated (1.22-fold over control values; P = 0.0006) and LH-stimulated (9.6-fold over control values; P = 0.007) MMPI expression and activity. Lastly, IL-1 beta and LH significantly (P < 0.05) decreased estrogen production by approximately 33% compared to that in cultures with LH only. It is concluded from the current study that IL-1 beta is a mediator of MMPI expression as well as granulosa cell steroidogenesis, and that this cytokine has divergent actions in the presence and absence of LH.

Animals↗

PubMed1995

Expression and activity of ovarian tissue inhibitors of metalloproteinases during pseudopregnancy in the rat.

The present study examined the role of tissue inhibitors of metalloproteinases (TIMPs) in tissue remodeling that occurs during luteal development and regression throughout pseudopregnancy in the rat. Pseudopregnancy was induced in immature female rats by eCG/hCG priming. Animals (n = 4 per time point) were killed on Days 1, 2, 4, 8, 12, 14, and 16 of pseudopregnancy (post hCG administration), and ovaries were removed and analyzed for metalloproteinase inhibitor activity or TIMP-1, TIMP-2, and TIMP-3 mRNA expression. Inhibitory activity was highest in Day-1 samples (41.35 +/- 6.50 inhibitory units), and inhibitor activity significantly decreased (p < 0.05) thereafter to minimal values at Day 12 (8.14 +/- 2.71 inhibitory units). Methylamine hydrochloride treatment, which inactivates macroglobulin-type inhibitors, revealed that the majority of the inhibitor activity in the Day-1 samples (82.6%) and the Day-16 samples (77.3%) could be attributed to TIMPs. To further distinguish the contribution of each TIMP to this activity, Northern analysis for TIMP-1, -2, and -3 was performed. Analysis of TIMP mRNA expression revealed that TIMP-1 transcript expression was highest (p = 0.00009) at Day 1, decreased approximately 3- to 20-fold from Days 2 to 12, respectively, and again increased at Days 14-16. However, TIMP-2 expression did not change (p > 0.05) over any of the time points studied. In contrast to TIMP-1 and TIMP-2 expression, TIMP-3 mRNA expression was lowest during Days 1 and 2 of pseudopregnancy, increased approximately 4-fold at Day 4, peaked at Day 8, and remained elevated throughout the remainder of pseudopregnancy.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1994

Detection of a unique 32-kd protein in the peritoneal fluid of women with endometriosis.

OBJECTIVE: To test the hypothesis that endometriotic tissue secretes endometriotic-specific proteins into the peritoneal fluid (PF) of women with endometriosis. DESIGN: A prospective design was utilized in this study. SETTING: Tertiary care, university-based center and reproductive endocrinology laboratory. PARTICIPANTS: Women of reproductive age who were infertile with endometriosis (n = 19), as well as without endometriosis (n = 7), and fertile women undergoing tubal ligation (n = 6). INTERVENTIONS: Collection of PF fluid via laparoscopy. MAIN OUTCOME MEASURES: Peritoneal fluid proteins were isolated and assessed by two-dimensional polyacrylamide gel electrophoresis. RESULTS: Two-dimensional electrophoresis of PF proteins isolated a group of proteins (M(r) = 32 to 40 kd, pI = 4.5 to 5.2) in all PF samples that was similar to the rat endometriotic implant-specific protein, Endo-1. This group of proteins consisted of 5 to 12 individual proteins with endometriosis PF containing a significantly higher number of proteins (median = 11) compared with either PF from infertile women without endometriosis (median = 8) or from women undergoing tubal ligation (median = 7). In addition, one protein (M(r) = 32 kd, pI = 5.8), termed EPF-32, was detected predominantly (18 of 19 samples analyzed) in PF from women with endometriosis. This protein was also detected in PF from infertile women without endometriosis (2 of 7 samples) but not in the PF of fertile women undergoing tubal ligation (0 of 6 samples). The appearance of this protein was not associated with the severity of endometriosis. CONCLUSION: It is concluded from this study that PF from women with endometriosis predominantly contains a 32-kd protein (EPF-32) compared with the PF of women without the disease. The role of EPF-32 in the pathophysiology of endometriosis is not established but this protein may function as a diagnostic marker for endometriosis.

Ascitic Fluid↗

PubMed1990

Interleukin-1 beta is a potent stimulator of prostaglandin synthesis in bovine luteal cells.

Interleukin-1 (IL-1) is a polypeptide that has both local and systemic effects on numerous tissues, including endocrine cells. To evaluate the effect of IL-1 on luteal function, bovine luteal cells were cultured for 5 days with increasing concentrations (0.1, 0.5, 1.0, 2.5, 5.0, 10.0 ng/ml) of recombinant bovine interleukin-1 beta (rbIL-1 beta). IL-1 beta increased the production of luteal 6-keto-prostaglandin-F1 alpha (6-keto-PGF1 alpha), prostaglandin E2 (PGE2), and prostaglandin F2 alpha (PGF2 alpha) in a dose-dependent manner, but had no effect on progesterone (P4) production. Treatment with the cyclooxygenase inhibitor, indomethacin (5 micrograms/ml), inhibited basal, as well as rbIL-1 beta-stimulated prostaglandin production. Addition of Iloprost (a synthetic analogue of prostacyclin, 5 ng/ml) suppressed basal production of PGF2 alpha and PGE2, but did not reduce the stimulatory effect of rbIL-1 beta. Similarly, PGF2 alpha suppressed basal, but not IL-1 beta-stimulated, production of 6-keto-PGF1 alpha. PGE2 had no effect on the synthesis of either PGF2 alpha or 6-keto-PGF1 alpha. P4 (1.75 micrograms/ml) reduced basal as well as rbIL-1 beta-stimulated production of 6-keto-PGF1 alpha, PGE2, and PGF2 alpha. These results indicate that IL-1 beta could serve as an endogenous regulator of luteal prostaglandin production. It appears that IL-1 beta action is not modified by exogenous prostaglandins, but is at least partially regulated by elevated P4. It is possible that the role of IL-1 beta in stimulation of luteal prostaglandin production may be confined to a period characterized by low P4 levels, such as during luteal development or regression.

6-Ketoprostaglandin F1 alpha↗

PubMedDate not supplied

Immunomodulation of rat endometriotic implant growth and protein production.

PROBLEM: The immune system has been implicated in the pathophysiology of endometriosis. To determine if modulation of the immune system influences endometriotic implant growth and protein production, the following experiment was conducted. METHOD: Female rats with surgically induced endometriosis were treated with either the immunosuppressive agent pentoxifylline (Pent; 5 mg/kg BW; N = 16) or a vehicle (N = 16) for 7 consecutive days, then killed. Twenty-four hours before death, 8 animals from each group were injected intraperitoneally with the immunostimulatory agent lipopolysaccharide (LPS; 200 micrograms/kg BW). At death, endometriotic implants were measured and protein production assessed by two-dimensional polyacrylamide gel electrophoresis. RESULT: Pentoxifylline significantly (P < 0.001) reduced endometriotic implant size (2.15 +/- 0.65 mm2 vs. 17.13 +/- 1.98 mm2) whereas LPS was without effect (18.32 +/- 2.57 mm2 vs. 17.13 +/- 1.98 mm2). Pentoxifylline also suppressed production of a portion of the proteins that comprise the implant specific group of proteins, ENDO-2, whereas LPS induced the production of two additional ENDO-2 proteins. CONCLUSION: Immunomodulatory agents can modulate rat endometriotic implant growth and production of implant-specific proteins.

Animals↗