transforming-growth-factor-beta and Peritoneal-Diseases

Endometriosis is estimated to affect 6-10% of women of reproductive age and it is associated with chronic pelvic pain, dysmenorrhoea and subfertility. It is currently managed surgically or medically but symptoms recur in up to 75% of cases and available medical treatments have undesirable side effects. Endometriosis is defined as the presence of endometrial tissue outside the uterus with lesions typically found on the peritoneum. The aetiology of endometriosis is uncertain but there is increasing evidence that transforming growth factor (TGF)-β plays a major role.. A descriptive review was undertaken of the published literature on the expression pattern of TGF-β ligands and signalling molecules in women with and without endometriosis, and on the potential roles of TGF-β signalling in the development and progression of peritoneal endometriosis. The current understanding of the TGF-β signalling pathway is summarized.. We searched the Pubmed database using the terms 'transforming growth factor beta' and 'endometriosis' for studies published between 1995 and 2016. The initial search identified 99 studies and these were used as the basic material for this review. We also extended our remit for important older publications. In addition, we searched the reference lists of studies used in this review for additional studies we judged as relevant. Studies which were included in the review focused on peritoneal endometriosis only as increasing evidence suggests that ovarian and deep endometriosis may have a differing pathophysiology. Thus, a final 95 studies were included in the review.. TGF-β1 is reported to be increased in the peritoneal fluid, serum, ectopic endometrium and peritoneum of women with endometriosis compared to women without endometriosis, and TGF-β1-null mice have reduced endometriosis lesion growth when compared to their wild-type controls. Studies in mice and women have indicated that increasing levels of TGF-β ligands are associated with decreased immune cell activity within the peritoneum, together with an increase in ectopic endometrial cell survival, attachment, invasion and proliferation, during endometriosis lesion development. TGF-β1 has been associated with changes in ectopic endometrial and peritoneal cell metabolism and the initiation of neoangiogenesis, further fuelling endometriosis lesion development.. Together these studies suggest that TGF-β1 plays a major role in the development of peritoneal endometriosis lesions and that targeting this pathway may be of therapeutic potential.

Topics: Endometriosis; Endometrium; Female; Humans; Peritoneal Diseases; Peritoneum; Transforming Growth Factor beta

Whether induced by infection, inflammation, ischemia, and/or surgical injury, peritoneal adhesions are the leading cause of pelvic pain, bowel obstruction, and infertility. Although some patients develop limited scar tissues, others for unknown reasons develop severe adhesions from seemingly equal procedures. Additionally in the same patient, adhesions develop at one surgical site but not in another. The mechanisms underlying the predisposition to form scars as well as their site specificity are unknown. Because a large number of intraperitoneal surgical procedures are performed each day, many patients are at risk of developing postoperative adhesions. As such, understanding the nature of molecular events and their mechanisms of action is essential, and in the absence of such information, attempts to prevent patients from developing adhesions will remain an empirical process. An unprecedented advancement in surgical techniques have resulted in minimizing peritoneal tissue injury that cause adhesion formation. Increased understanding of the cellular and molecular events that lead to scar tissue formation has also led to the identification of many biologically active molecules with the potential of regulating inflammatory and immune responses, angiogenesis, and tissue remodeling, events that are central to normal peritoneal wound healing and adhesion formation. This article attempts to highlight some of the key molecules (i.e., the transforming growth factor family and its regulatory mechanisms) that are recognized to regulate peritoneal wound repair and adhesion formation. Such understanding of peritoneal biology not only will assist us to better manage patients with adhesions but also will assist those with endometriosis and malignant diseases that affect the peritoneal cavity.

Topics: Animals; Female; Fibrosis; Humans; Models, Biological; Peritoneal Diseases; Peritoneum; Protein Binding; Tissue Adhesions; Transforming Growth Factor beta; Wound Healing

We have provided evidence from adenovirus-mediated gene transfer to the peritoneum that transforming growth factor-beta1 mimics many of the functional and structural changes in the peritoneum in patients on long-term peritoneal dialysis, including fibrosis, increased submesothelial thickness, angiogenesis, increased solute transport, and ultrafiltration dysfunction. We review several key properties of this important fibrogenic molecule.

Topics: Humans; Peritoneal Diseases; Peritoneum; Transforming Growth Factor beta; Transforming Growth Factor beta1

Peritoneal adhesions are a major problem for health and economy. An adequate and atraumatic surgical technique is essential in the prevention of peritoneal adhesions. Laparoscopic microsurgery should be developed and promoted. The following conclusions can be proposed about the treatments available Adequately designed clinical studies are too rare. Future studies are necessary to obtain information on pregnancy and pain as primary outcomes. Cost implications of adhesion prevention treatments also have to be addressed.

Topics: Animals; Epithelium; Humans; Hyaluronic Acid; Laparoscopy; Microsurgery; Peritoneal Diseases; Postoperative Complications; Sutures; Therapeutic Irrigation; Tissue Adhesions; Transforming Growth Factor beta

To examine the effects of the PROACT treatment on the fibrinolytic system and inflammatory cytokines in human peritoneum.. Controlled clinical study.. University hospital.. Nine subjects undergoing laparotomy had peritoneal samples taken at the incision.. The PROACT applicator was inserted through the peritoneal incision, and treatment of peritoneum was performed twice. A peritoneal sample was taken from one treated area. At closure, the second treated sample and an additional control sample were taken. All four samples were snap frozen in liquid nitrogen. Samples were homogenized and protein content extracted.. Concentrations of total and active transforming growth factor-beta 1 (TGF-beta1), tumor necrosis factor-alpha (TNF-alpha), tissue-type plasminogen activator (t-PA), urokinase plasminogen activator (uPA), and plasminogen activator inhibitor 1 (PAI-1) were obtained.. Total TGF-beta1 at opening was 30% less in treated samples. At closure, active TGF-beta1 increased significantly (163%) in control samples and not in treated samples. Tumor necrosis factor alpha was detectable only in control samples at closure. During surgery, tPA levels showed a marked decrease in control samples vs. a small increase in treated samples. Levels of uPA increased significantly only in the control samples. In control samples, tPA/PAI-1 ratio was two thirds of treated sample ratio.. Heating of the peritoneum with the PROACT System modulates the biologic tissue response to induce effects that would be consistent with inhibition of postoperative adhesion development.

Topics: Aged; Aged, 80 and over; Female; Humans; Hyperthermia, Induced; Laparotomy; Male; Middle Aged; Peritoneal Diseases; Plasminogen Activator Inhibitor 1; Tissue Adhesions; Tissue Plasminogen Activator; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Urokinase-Type Plasminogen Activator

Adhesion formation that occurred after alkali-induced injury of the cecum was used as a novel adhesion model in rats, and it was compared with that of a common adhesion model after abrading the cecum. Using the novel adhesion model, inhibition of adhesion formation by a chymase inhibitor, Suc-Val-Pro-PheP(OPh)2, and by sodium hyaluronate/carboxymethylcellulose (Seprafilm) was evaluated, and their mechanisms were assessed. The degree of adhesion formation was more severe and more stable in the alkali-induced injury model than in the abrasion-induced injury model. Both the chymase inhibitor and Seprafilm showed significant attenuation of the degree of adhesion 14 days after alkali-induced injury. Chymase activity in the cecum was significantly increased after alkali-induced injury, but it was significantly attenuated by the chymase inhibitor and Seprafilm. Myeloperoxidase and transforming-growth factor (TGF)-β levels were significantly increased after alkali-induced injury, but they were attenuated by both the chymase inhibitor and Seprafilm. At the level of the adhesions, the numbers of both chymase-positive cells and TGF-β-positive cells were significantly increased, but their numbers were reduced by the chymase inhibitor and Seprafilm. In conclusion, a chymase inhibitor attenuated the degree of adhesions to the same degree as Seprafilm in a novel peritoneal adhesion model that was more severe and more stable than the common adhesion model, and not only the chymase inhibitor, but also Seprafilm reduced the chymase increase at the adhesions.

Topics: Animals; Carboxymethylcellulose Sodium; Cecum; Chymases; Disease Models, Animal; Gene Expression; Hyaluronic Acid; Male; Peritoneal Diseases; Peroxidase; Protease Inhibitors; Rats; Rats, Sprague-Dawley; Tissue Adhesives; Transforming Growth Factor beta

Peritoneal fibrosis is a serious complication of peritoneal dialysis (PD). It has been reported that administration of mizoribine, an effective immunosuppressant, ameliorated renal fibrosis in a rat model of unilateral ureteral obstruction. We therefore examined the effects of mizoribine in an experimental model of peritoneal fibrosis.. 24 rats were given a daily intraperitoneal injection of chlorhexidine gluconate and ethanol dissolved in saline. The rats were divided into three groups (n = 8 per group) that received either vehicle or mizoribine at a dose of 2 or 8 mg/kg once a day. 28 days after the start of the treatments the rats were sacrificed and peritoneal tissue samples collected. Macrophage infiltration (ED1), myofibroblast accumulation (alpha-smooth muscle actin (SMA)) and expression of type III collagen, transforming growth factor (TGF)-beta and monocyte chemotactic protein-1 (MCP-1) were examined by immunohistochemistry.. Mizoribine significantly suppressed submesothelial zone thickening and reduced macrophage infiltration. Mizoribine also reduced collagen III(+) area and decreased the number of alpha-SMA(+), TGF-beta(+) and MCP-1(+) cells. The magnitude of the changes observed was dose-dependent.. The administration of mizoribine prevented the progression of peritoneal fibrosis in this rat model. Mizoribine may represent a novel therapy for peritoneal sclerosis in patients undergoing long-term PD.

Topics: Actins; Animals; Cell Movement; Chemokine CCL2; Chlorhexidine; Collagen Type III; Disease Models, Animal; Disease Progression; Dose-Response Relationship, Drug; Fibrosis; Immunosuppressive Agents; Macrophages, Peritoneal; Peritoneal Diseases; Peritoneum; Rats; Rats, Wistar; Ribonucleosides; Transforming Growth Factor beta

Postoperative intra-abdominal adhesion is associated with high morbidity and mortality. Smad7, a protein that occupies a strategic position in fibrogenesis, inhibits the transforming growth factor (TGF) beta/Smad signalling pathway. In this study the therapeutic potential of exogenous Smad7 in preventing fibrogenesis in postoperative intra-abdominal adhesion was investigated.. Intra-abdominal adhesion was induced in a rodent model by peritoneal abrasion. Smad7 was delivered into the peritoneal cavity by a non-viral ultrasound-microbubble-mediated naked gene transfection system. The effect of Smad7 transgene on adhesion formation was studied by measuring changes in TGF-beta, fibrogenic factors, alpha-SMA and Smad2/3 activation in the anterior abdominal wall.. Four weeks after surgical abrasion, all rats developed significant peritoneal adhesion with enhanced TGF-beta expression, increased levels of extracellular matrix components and activated myofibroblasts, accompanied by decreased Smad7 expression and increased Smad2/3 activation. In rats treated with the Smad7 transgene, the incidence and severity of peritoneal adhesion were significantly reduced, with biochemical downregulation of fibrogenic factors and inhibition of Smad2/3 activation. Serial quantitation using magnetic resonance imaging revealed a significant reduction in adhesion areas from day 14 onwards.. Ultrasound-microbubble-mediated gene transfection provides timely targeted gene delivery for the treatment of postoperative peritoneal adhesions.

Topics: Abdominal Wall; Animals; Extracellular Matrix; Gene Transfer Techniques; Genetic Vectors; Immunohistochemistry; Male; Microbubbles; Peritoneal Diseases; Rats; Rats, Sprague-Dawley; Reverse Transcriptase Polymerase Chain Reaction; Smad2 Protein; Smad3 Protein; Smad7 Protein; Tissue Adhesions; Transforming Growth Factor beta; Transgenes; Up-Regulation

Transforming growth factor beta (TGF-beta) plays a critical role in the pathogenesis of the peritoneal fibrosis that complicates long-term peritoneal dialysis (PD). We studied the TGF-beta/Smad signaling pathway in peritoneal fibrosis induced in uremic rats treated with PD and explored the therapeutic potential of Smad7 to prevent fibrogenesis. After subtotal nephrectomy, uremic rats were treated with peritoneal dialysis using 4.25% dextrose-containing fluid. The peritoneum of uremic rats treated with PD demonstrated fibrosis, increased TGF-beta expression, increased Smad2/3 activation, decreased Smad7 expression, and increased expression of fibrogenic and angiogenic factors. In addition, peritoneal function was impaired and its structure was altered, including a thickened submesothelial layer. In rats transfected with a Smad7 transgene using an ultrasound-microbubble-mediated system, peritoneal fibrosis was attenuated, peritoneal function was improved, and Smad2/3 activation was inhibited. We suggest that administration of Smad7 inhibits peritoneal fibrogenesis in uremic rats treated with PD by correcting the imbalance between downregulated Smad7 and activated Smad2/3. Blockade of the TGF-beta/Smad signaling pathway may represent a novel therapeutic approach to prevent peritoneal fibrosis in patients treated with PD.

Topics: Animals; Extracellular Matrix; Fibrosis; Gene Expression; Genetic Therapy; Male; Neovascularization, Pathologic; Peritoneal Dialysis, Continuous Ambulatory; Peritoneal Diseases; Peritoneum; Rats; Rats, Sprague-Dawley; Signal Transduction; Smad7 Protein; Transfection; Transforming Growth Factor beta; Transgenes; Uremia

Injury to the peritoneum during surgery is followed by a healing process that frequently results in the attachment of adjacent organs by a fibrous mass, referred commonly as adhesions. Because injuries to the peritoneum during surgery are inevitable, it is imperative that we understand the mechanisms of adhesion formation to prevent its occurrence. This requires thorough understanding of the molecular sequence that results in the attachment of injured peritoneum and the development of fibrous tissue. Recent data show that fibroblasts from the injured peritoneum may play a critical role in the formation of adhesion tissues. Therefore, identifying changes in gene expression pattern in the peritoneal fibroblasts during the process may provide clues to the mechanisms by which adhesion develop.. In this study, we compared expression patterns of larger number of genes in the fibroblasts isolated from adhesion and normal human peritoneum using gene filters. Contributions of TGF-beta1 and hypoxia in the altered expression of specific genes were also examined using a semiquantitative RT-PCR technique.. Results show that several genes are differentially expressed between fibroblasts of normal and adhesion peritoneum and that the peritoneal fibroblast may acquire a different phenotype during adhesion formation. Genes that are differentially expressed between normal and adhesion fibroblasts encode molecules involved in cell adhesion, proliferation, differentiation, migration and factors regulating cytokines, transcription, translation and protein/vesicle trafficking.. Our data substantiate that adhesion formation is a multigenic phenomenon and not all changes in gene expression pattern between normal and adhesion fibroblasts are the function of TGF-beta1 and hypoxia that are known to influence adhesion formation. Analysis of the gene expression data in the perspective of known functions of genes connote to additional targets that may be manipulated to inhibit adhesion development.

Topics: Cell Hypoxia; DNA, Complementary; Female; Fibroblasts; Gene Expression Profiling; Gene Expression Regulation; Humans; Peritoneal Diseases; Peritoneum; RNA, Messenger; Tissue Adhesions; Transforming Growth Factor beta; Transforming Growth Factor beta1

Peritoneal fibrosis (or sclerosis) is a complication of peritoneal dialysis (PD) and includes a wide spectrum of peritoneal structural changes, ranging from mild inflammation to severe sclerosing peritonitis and encapsulating-sclerosing peritonitis. In parallel with fibrosis, the peritoneum shows a progressive increase in capillary number (angiogenesis) and vasculopathy, which are involved in both the elevation of small solute transport across the peritoneal membrane and ultrafiltration failure. The most important substances from the PD solutions responsible for peritoneal deterioration are glucose and glucose degradation products, which stimulate transforming growth factor (TGF)beta and vascular endothelial growth factor (VEGF) production by mesothelial cells (MCs). TGFbeta is a potent pro-fibrotic factor and induces epithelial-mesenchymal transition (EMT) of the MC. Local production of VEGF during PD appears to play a central role in the processes leading to peritoneal neo-angiogenesis and functional decline. This review discusses the mechanisms implicated in peritoneal structural alteration and points to EMT of MC as the protagonist and initiator of peritoneal membrane injury, through an increment of the submesothelial fibroblast population. We also propose possible mechanisms of regulation and new targets for inhibition of EMT.

Topics: Epithelium; Fibroblasts; Fibrosis; Glucose; Humans; Models, Biological; Neovascularization, Pathologic; Peritoneal Dialysis; Peritoneal Diseases; Peritoneum; Transforming Growth Factor beta; Vascular Endothelial Growth Factor A

Transforming growth factor-beta (TGF-beta), a multifunctional growth factor, represents three mammalian isoforms, TGF-beta1, TGF-beta2, and TGF-beta3. In cutaneous wound healing, combined neutralization of TGF-beta1 and -beta2 or addition of TGF-beta3 reduces scar formation. Here, we investigated whether experimental manipulation of TGF-beta isoforms reduced adhesion formation after injury to the peritoneum. Adhesions were produced in mice by surgical abrasion of adjacent serosa followed by close apposition. In the first part of this study, a detailed analysis of TGF-beta isoform distribution was performed through immunolocalization. TGF-beta isoforms clearly showed a unique temporal and spatial pattern of expression after peritoneal wounding. Based on this pharmacokinetic data, we next administered neutralizing antibodies to TGF-beta1 and -beta2 or exogenous TGF-beta3 peptide by local application and intraperitoneal injection at various times before and after surgery. At day 7 after surgery, addition of neutralizing antibodies to both TGF-beta1 and -beta2 significantly reduced the number and size of adhesions (P < 0.05) compared with the vehicle control. By contrast, exogenous addition of TGF-beta3 either had no effect or increased adhesion formation compared to the vehicle control. In conclusion, these results show that by blocking both TGF-beta1 and TGF-beta2 using neutralizing antibodies, it is possible to prevent abdominal adhesion formation.

Topics: Animals; Antibodies; Ascitic Fluid; Cecum; Immunohistochemistry; Kinetics; Male; Mice; Mice, Inbred C57BL; Models, Anatomic; Neutralization Tests; Peritoneal Diseases; Peritoneal Lavage; Protein Isoforms; Random Allocation; Tissue Adhesions; Tissue Distribution; Transforming Growth Factor beta; Wound Healing

Peritoneal matrix accumulation is a major characteristic of encapsulating peritoneal sclerosis (EPS), which is a serious complication in long-term peritoneal dialysis (PD) patients. We reported previously that TGF-beta stimulates collagen gene expression in cultured HPMC, and is attenuated by pentoxifylline (PTX). The SMAD family and the mitogen-activated protein kinase (MAPK) (ERK1/2, JNK and p38(HOG)) pathways have been shown to participate in TGF-beta signalling. However, how PTX modulates the intracellular signalling downstream to TGF-beta remains undetermined in HPMC. In this study, we explored these signalling pathways in HPMC, and investigated the molecular mechanisms involved in the inhibitory effects of PTX on TGF-beta-induced collagen gene expression in HPMC.. HPMC was cultured from human omentum by an enzyme digestion method. The expression of collagen alpha1(I) mRNA was determined by northern blotting, while the SMAD proteins and the MAPK kinase activity were determined by western blotting.. TGF-beta-stimulated collagen alpha1(I) mRNA expression of HPMC was inhibited by PTX. The Smad2, ERK1/2 and p38(HOG) pathways were activated in response to TGF-beta. However, TGF-beta displayed no activation of the JNK pathway in HPMC. The addition of PD98059 and SB203580, which blocked the activation of ERK1/2 and p38(HOG), respectively, suppressed the TGF-beta-induced collagen alpha1(I) mRNA expression. At a concentration (300 micro g/ml) that inhibited the collagen gene expression, PTX suppressed the ERK1/2 and p38(HOG) activation by TGF-beta. In contrast, PTX had no effect on the TGF-beta-induced activation of Smad2, under the same concentration.. PTX inhibits the TGF-beta-induced collagen gene expression in HPMC through modulating the ERK1/2 and p38(HOG) pathways. Our study of PTX may provide the therapeutic basis for clinical applications in the prevention of EPS.

Topics: Blotting, Northern; Blotting, Western; Cells, Cultured; Epithelial Cells; Humans; Pentoxifylline; Peritoneal Cavity; Peritoneal Diseases; Peritoneum; Probability; RNA, Messenger; Sclerosis; Sensitivity and Specificity; Signal Transduction; Trans-Activators; Transforming Growth Factor beta

Mast cells are closely related to adhesion formation, while it has been unclear which factor in mast cells plays an important role in the development of adhesion formation. To clarify the role of chymase produced from mast cells in adhesion formation, we investigated the preventive effect of a specific chymase inhibitor, BCEAB, on adhesion formation in a hamster experimental model.. Hamsters were administered orally once daily with 100 mg/kg of BCEAB or placebo from the operated day to 1 week after the operation. The uterus was grasped and denuded by a swab.. One week after the operation, the scores for adhesion formation in the chymase inhibitor-treated group were significantly decreased in comparison with those in the placebo-treated group (placebo-treated group, 2.80 +/- 0.20; chymase inhibitor-treated group 1.60 +/- 0.31: P < 0.01). The chymase activity in the injured uterus was also significantly suppressed in the chymase inhibitor-treated group (placebo-treated group, 17.3 +/- 2.69 mU/mg protein; chymase inhibitor-treated group 9.60 +/- 0.89: P < 0.05). After scraping the utelus, the level of transforming growth factor-beta in the peritoneal fluid was significantly increased in the placebo-treated group, while it was suppressed to 70% by the treatment with BCEAB.. The specific chymase inhibitor BCEAB may be a useful drug for prevention of adhesion formation.

Topics: Animals; Azetidines; Benzoates; Chymases; Cricetinae; Female; Mesocricetus; Peritoneal Diseases; Serine Endopeptidases; Tissue Adhesions; Transforming Growth Factor beta

Little is known about the role of peritoneal fibroblasts in adhesion formation. This study determines the effect of hypoxia and transforming growth factor (TGF)-beta1 treatment on the expression of TGF-beta1-3 and TGF-betaI and betaII receptors in human peritoneal fibroblasts (HPF). TGF-beta isoforms and their receptors have been implicated as mediators of the healing process and adhesion development.. HPF were cultured under normal and hypoxic condition, and treated with and without (1 ng/mL) TGF-beta1 for 24 hr. Total RNA from each group was subjected to multiplex reverse transcriptase-polymerase chain reaction (RT/PCR) to quantitate TGF-beta1-3 and TGF-betaI and betaII receptors messenger RNA (mRNA) levels.. Hypoxia resulted in a significant increase in TGF-beta1 (26%; P < 0.05), TGF-betaIR (34%; P < 0.05) and TGF-betaIIR (29%; P < 0.05) mRNA levels, with no effect on TGF-beta2 or beta3. TGF-beta1 treatment resulted in a significant increase in TGF-beta1 (35%; P < 0.05), but a decrease in TGF-beta2 (22%; P < 0.05) and no effect on TGF-beta3, TGF-betaIR or TGF-betaIIR. Combined treatment of hypoxia and TGF-beta1 caused a significant increase in TGF-beta1 (37%; P < 0.05), TGF-beta2 (12%; P < 0.05), TGF-betaIR (32%; P < 0.05) and TGF-betaIIR (34%; P < 0.05). There is no significant change in the mRNA levels of TGF-beta3 in any of the treatments.. Hypoxia and TGF-beta1 treatments of cultured HPF modulate the expression of TGF-beta1, beta2 and beta3 and their receptors betaIR and betaIIR by increasing the ratio of TGF-beta1 and beta2 to beta3, thus favoring the development of peritoneal adhesion.

Topics: Activin Receptors, Type I; Cell Hypoxia; Cells, Cultured; Fibroblasts; Gene Expression Regulation; Humans; Peritoneal Diseases; Peritoneum; Protein Serine-Threonine Kinases; Receptor, Transforming Growth Factor-beta Type I; Receptor, Transforming Growth Factor-beta Type II; Receptors, Transforming Growth Factor beta; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Tissue Adhesions; Transforming Growth Factor beta; Transforming Growth Factor beta1; Transforming Growth Factor beta2; Transforming Growth Factor beta3; Wound Healing

Elevated local expression of transforming growth factor (TGF-beta) has been associated with increased incidence of peritoneal adhesion formation. In this study we determine whether differences in basal expression of TGF-beta in serosal tissue of peritoneal organs correlate with incidence of adhesion formation. Serosal tissue of parietal peritoneum, uterus, oviduct, ovary, omentum, large and small bowels as well as adhesions, skin, fascia, subcutaneous tissue, peritoneal fluid and serum were collected from 57 subjects with/without adhesions who were undergoing abdominal/pelvic surgery. To determine TGF-beta1 and TGF-beta3 mRNA and protein expression, total RNA and protein were isolated from these tissues and along with the fluids, subjected to quantitative RT-PCR and enzyme-linked immunosorbent assay (ELISA) respectively. Tissue sections were immunostained for TGF-beta1 and TGF-beta3 protein. We found that TGF-beta1 and TGF-beta3 mRNA and protein are expressed in these tissues and present in peritoneal fluids and serum, with considerable variations in level of their expression. Comparatively, there was more variation in TGF-beta1 than TGF-beta3 expression without age or gender relation. Adhesions express a significantly higher TGF-beta1 mRNA and have the highest TGF-beta1:TGF-beta3 ratio, with lowest concentrations and ratio detected in omentum, small and large bowels; in contrast uterus expresses higher TGF-beta3, with lowest concentrations detected in subcutaneous tissue and large bowels (P < 0.05). A similar trend was also observed for total (active + latent) TGF-beta1 protein expression, with low active TGF-beta1 that was not significantly different among the tissue extracts and fluids. However, the lowest active:total TGF-beta1 ratio was found in adhesions and ovary. In subjects with adhesions, the adhesions express significantly more TGF-beta1 compared to parietal peritoneum (P < 0.05). Immunoreactive TGF-beta1 and TGF-beta3 protein were present in various cell types in these tissues with intensity reflecting their mRNA and protein expression. In conclusion, we provided evidence that serosal tissue of various peritoneal organs and adhesions express TGF-beta1 and TGF-beta3. Since TGF-beta is expressed differently in these tissues and tissue injury often alters the expression of TGF-beta, we propose that tissues with a higher basal expression of TGF-beta may become predisposed to develop more adhesions compared to others.

Topics: Adult; Aged; Aged, 80 and over; Ascitic Fluid; Enzyme-Linked Immunosorbent Assay; Fallopian Tubes; Fascia; Female; Gene Expression; Humans; Intestines; Male; Middle Aged; Omentum; Ovary; Peritoneal Diseases; Peritoneum; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Skin; Tissue Adhesions; Transforming Growth Factor beta; Transforming Growth Factor beta1; Transforming Growth Factor beta3; Uterus

Continuous ambulatory peritoneal dialysis (CAPD) has emerged as an important dialysis treatment modality worldwide. One of the major complications is bacterial peritonitis, which may result in subsequent technique failure because of loss of peritoneal clearance or peritoneal fibrosis. Bacterial peritonitis leads to the release of proinflammatory cytokines from resident and infiltrating cells in the peritoneal cavity. We studied 35 patients undergoing CAPD with acute bacterial peritonitis. All patients treated with antibiotics for 2 weeks after the clinical diagnosis of peritonitis had a good recovery. Peritoneal dialysate effluent (PDE) was collected on days 1, 3, 5, 10, 21, and 42 after the start of treatment. Cell populations were monitored by flow cytometry. PDE levels of interleukin-1beta (IL-1), IL-6, transforming growth factor-beta (TGF-beta), and basic fibroblast growth factor (FGF) were measured by enzyme-linked immunosorbent assay. Gene transcription of TGF-beta in macrophages from PDE was measured by quantitative polymerase chain reaction. Bacterial peritonitis was associated with a sharp increase in total cell and neutrophil counts (400-fold) in PDE up to 3 weeks after peritonitis despite clinical remission (P < 0.0001). There was an increased absolute number of macrophages during the first 3 weeks despite the reduced percentage of macrophages among total cells in PDE compared with noninfective PDE. There was a progressive increase in the percentage of mesothelial cells or dead cells in the total cell population in PDE over the entire 6-week period. PDE levels of IL-1, IL-6, TGF-beta, and FGF increased markedly on day 1 before their levels decreased gradually. PDE levels of these cytokines or growth factors were significantly greater than those in noninfective PDE (n = 76) throughout the study period (P < 0.01). Similarly, TGF-beta complementary DNA (cDNA) molecules per macrophage were significantly greater than those of macrophages in noninfective PDE throughout this period (P < 0.01). There was no significant correlation between PDE levels of TGF-beta and TGF-beta cDNA molecules per macrophage, suggesting that peritoneal macrophages are not the only source of TGF-beta in PDE. We conclude there is an active release of proinflammatory cytokines and sclerogenic growth factors through at least 6 weeks despite apparent clinical remission of peritonitis. The peritoneal cytokine networks after peritonitis may potentially affect the physiological pro

Topics: Bacterial Infections; Cytokines; DNA, Complementary; Female; Fibroblast Growth Factor 2; Fibrosis; Flow Cytometry; Humans; Interleukin-1; Interleukin-6; Macrophages, Peritoneal; Male; Middle Aged; Peritoneal Dialysis, Continuous Ambulatory; Peritoneal Diseases; Peritonitis; Polymerase Chain Reaction; Transforming Growth Factor beta

The development of postoperative intraperitoneal adhesions continues to be a major concern for surgeons. The purpose of this study was to establish a postoperative adhesion model in rats, and to assess the effectiveness of tranilast (N-(3',4'-dimethoxycinnamoyl)anthranilic acid) in preventing postoperative adhesion formation. The adhesion model was established in 12 male Donryu rats. This involved two essential factors, drying and bleeding. Another 22 male Donryu rats were used to study the prevention of intraperitoneal adhesions. Tranilast was administered orally pre- and postoperatively. Adhesion strength was evaluated by grading, and basic fibroblast growth factor (bFGF) and transforming growth factor-beta-1 (TGF-beta1) concentration were measured. Postoperative intraperitoneal adhesions were seen in all rats, but the adhesions in the tranilast group were significantly less severe than those in the control group. Serum bFGF and TGF-beta1 levels in the tranilast group were lower at the time of surgery than those in the control group, and bFGF levels were lower at the endpoint of this study in the tranilast group than in the control group. The TGF-beta1 levels at the end-point did not differ between the two groups. These findings demonstrated that tranilast significantly reduced postoperative intraperitoneal adhesion formation.

Topics: Abdomen; Animals; Enzyme-Linked Immunosorbent Assay; Fibroblast Growth Factor 2; Male; ortho-Aminobenzoates; Peritoneal Diseases; Postoperative Complications; Rats; Rats, Inbred Strains; Statistics, Nonparametric; Tissue Adhesions; Transforming Growth Factor beta

Long-term influence of continuous ambulatory peritoneal dialysis (CAPD) on concentrations of transforming growth factor beta1 (TGF-beta1) and basic fibroblast growth factor (bFGF) in the peritoneal effluent, and the effect of peritonitis on these cytokines were investigated. TGF-beta1 and bFGF were assayed in effluent samples from dialysate bags collected during the initial week of treatment with CAPD and at 5 months. To determine the effect of peritonitis, dialysate bags were collected on admission to the hospital and on days 3 and 10 and also during non-infected steady state. Serum was drawn prior to infection and on days 1 and 10. TGF-beta1 increased more than threefold during the longitudinal follow-up period, median concentrations of 35 pg/ml to 106 pg/ml (P<0.05). No change in bFGF was seen during this initial 5 months. TGF-beta1 was increased on the first day of peritonitis (median concentration 169 pg/ml) and reached its maximum on day 3 of infection, (median concentration 216 pg/ml) (P<0.05 vs non-infected state, median concentration 39 pg/ml). Basic FGF reached a maximum on day three of infection (median concentration 7.7 pg/ml; P=0.01 vs non-infected state) and then slowly declined. In conclusion, TGF-beta1 is influenced by CAPD treatment per se, and together with bFGF is increased during peritonitis, indicating its importance in the peritoneum and its potential involvement in the development of tissue fibrosis and eventually ultrafiltration failure.

Topics: Adult; Aged; Corynebacterium; Dialysis Solutions; Escherichia coli; Female; Fibroblast Growth Factor 2; Fibrosis; Humans; Interleukin-13; Interleukin-2; Longitudinal Studies; Male; Middle Aged; Peritoneal Dialysis, Continuous Ambulatory; Peritoneal Diseases; Peritonitis; Staphylococcus aureus; Staphylococcus epidermidis; Streptococcus sanguis; Transforming Growth Factor beta; Uremia

Transforming growth factor beta (TGF-beta) is a major secretory product of macrophages which, through autocrine/paracrine pathways, play a central role in normal reproductive tissues as well as in disorders such as endometriosis and intraperitoneal adhesion formation. Using TGF-beta antisense oligonucleotides and U937 cells (a promonocytic human cell line) as an in-vitro model, the present study examined the autocrine mediated action of TGF-beta 1 on proliferation, anchor-dependent and -independent cell aggregation and expression of several mRNAs of cell surface adhesion molecules including integrins and platelet-endothelial cell adhesion molecule (PECAM-1). Northern blot analysis and enzyme-linked inmmunosorbent assay (ELISA) revealed that treatment with TGF-beta 1 antisense, but not sense or nonsense oligomers, in a dose-dependent manner (0.1-10 microM) down-regulated the expression of TGF-beta 1 mRNA and protein to undetectable amounts at the highest antisense concentration. TGF-beta 1 antisense at < 1 mM slightly increased, while at > 3 microM significantly inhibited, the rate of DNA synthesis and proliferation of these cells (P < 0.05). Treatment with TGF-beta 1 antisense promoted cell aggregation under anchor-independent culture conditions (plastic dishes), while it suppressed colony formation under anchor-dependent culture conditions (soft agar assay). U937 cells expressed alpha 2, alpha 3, alpha 4, alpha 5, alpha 6, beta 1 and beta 2 integrin mRNA and PECAM-1 mRNA, while alpha v, beta 3 and beta 5 integrin mRNA was undetectable. The relative amount of alpha 2, alpha 3, alpha 4, alpha 6, beta 1 and beta 2 integrin and PECAM-1 mRNA expression were down-regulated in a dose-dependent manner after TGF-beta 1 antisense treatment, while alpha 5 integrin mRNA expression was up-regulated, although it was undetectable at 10 microM antisense. In contrast, TGF-beta 1 antisense up-regulated beta 3 mRNA expression with maximal effect occurring at 10 microM. These results provide evidence that the autocrine loop of monocyte/macrophage-derived TGF-beta 1 action is essential for regulation of growth, aggregation and the expression of adhesion molecules by these cells. We propose that in disorders such as endometriosis and peritoneal fibrous adhesions, significantly higher numbers of tissue macrophages with the capacity to express excess TGF-beta 1 yield an environment able to promote cell-cell and cell-matrix interactions, and thus lead to further complications fr

Topics: Base Sequence; Cell Adhesion; Cell Aggregation; Cell Division; Cell Line; DNA; Endometriosis; Female; Gene Expression; Humans; Integrins; Monocytes; Oligonucleotides, Antisense; Peritoneal Diseases; Platelet Endothelial Cell Adhesion Molecule-1; RNA, Messenger; Tissue Adhesions; Transforming Growth Factor beta

To investigate the presence and cellular distribution of transforming growth factor (TGF)-beta s in surgically induced pelvic fibrous adhesions in rat uterine horns subjected to burn, crush, and debridement injury.. Thirty injured and 20 uninjured rats were treated postoperatively with intraperitoneal administration of either 2 micrograms/mL of recombinant human TGF-beta, 10 micrograms/mL TGF-beta neutralizing antibody, or phosphate-buffered saline + 500 micrograms rat serum albumin for 5 consecutive days. The intact (uninjured) and fibrous tissues were analyzed immunohistochemically for the presence of TGF-beta s using polyclonal antibodies to TGF-beta s 1-3.. The intact peritoneum immunostained with a lower intensity than fibrous adhesive tissues for TGF-beta 1, TGF-beta 2, and TGF-beta 3. The immunoreactive TGF-beta s were present in fibroblasts, inflammatory cells infiltrated into the fibrous adhesion, and endothelial and smooth-muscle cells of the arterioles. In the uterine tissue at the site of injury, the following immunostained for TGF-beta s: uterine serosal tissue, myometrial smooth-muscle cells, endometrial luminal and glandular epithelial cells, and inflammatory cells. However, endometrial stromal cells did not immunostain for TGF-beta s. There were no substantial differences in immunostaining intensities of fibrous adhesive tissues in the TGF-beta group, neutralizing TGF-beta antibody group, and the controls.. The data suggest that TGF-beta s may play a role in the formation and maintenance of fibrous adhesions following intraperitoneal injury.

Topics: Animals; Female; Peritoneal Diseases; Rats; Rats, Sprague-Dawley; Tissue Adhesions; Transforming Growth Factor beta