RESUMEN
Plasma concentration of lysophosphatidylcholine (LPC) was reported to decrease in patients with sepsis. However, the mechanisms of sepsis-induced decrease in plasma LPC levels are not currently well known. In mice subjected to cecal ligation and puncture (CLP), a model of polymicrobial peritoneal sepsis, we examined alterations in LPC-related metabolic parameters in plasma, i.e., the plasma concentration of LPC-related substances (i.e., phosphatidylcholine (PC) and lysophosphatidic acid (LPA)), and activities or levels in the plasma of some enzymes that can be involved in the regulation of plasma LPC concentration (i.e., secretory phospholipase A2 (sPLA2), lecithin:cholesterol acyltransferase (LCAT), acyl-CoA:lysophosphatidylcholine acyltransferase (LPCAT), and autotaxin (ATX)), as well as plasma albumin concentration. We found that levels of LPC and albumin and enzyme activities of LCAT, ATX, and sPLA2 were decreased, whereas levels of PC, LPA, and LPCAT1-3 were increased in the plasma of mice subjected to CLP. Bacterial peritonitis led to alterations in all the measured LPC-related metabolic parameters in the plasma, which could potentially contribute to sepsis-induced decrease in plasma LPC levels. These findings could lead to the novel biomarkers of sepsis.
Asunto(s)
Lisofosfatidilcolinas/metabolismo , Sepsis/sangre , Aciltransferasas/metabolismo , Animales , Modelos Animales de Enfermedad , Lisofosfatidilcolinas/sangre , Lisofosfolípidos/metabolismo , Metabolismo , Ratones , Enfermedades Peritoneales/sangre , Enfermedades Peritoneales/enzimología , Enfermedades Peritoneales/metabolismo , Fosfolipasas/metabolismo , Sepsis/enzimología , Sepsis/metabolismoRESUMEN
Endometriosis is a common reproductive disorder believed to be associated with matrix metalloproteinases (MMPs) activities for invasion and remodeling of endometrial tissues. Ectopic endometrium has higher capacity to produce proMMP-2 than eutopic tissues; however, the role of MMP-2 during early phase of endometriosis development is still unclear. In the present study, we investigated the role of MMP-2 in establishment and development of endometriosis in mouse model. The effect of curcumin on regression of endometriosis through protease/antiprotease balance between MMP-2 and TIMP-2 was also examined. After endometrial inoculation into peritoneum, we observed a significant elevation of proMMP-2 activity from day 2 onwards. This increased MMP-2 activity was associated with decreased expression of tissue inhibitor of MMP (TIMP)-2, while a significant up-regulation of active MMP-2 activity was observed from day 3 onwards. The activation of proMMP-2 to active MMP-2 was associated with increased expression of membrane type 1 matrix metalloproteinase (MT1MMP). Curcumin at a dose of 48 mg/kg b.w. repressed the MMP-2 activity via up-regulation of bound TIMP-2 expression, thus delayed endometriosis development. In addition, curcumin inhibited production of active MMP-2 by down-regulating MT1MMP expression. Moreover, endometriotic progression was directly linked with increased MMP-2/TIMP-2 ratio which was delayed by curcumin pretreatment. In summary, our study documents the regulation of MMP-2 activity by TIMP-2 during the early phase of endometriosis development and inhibitory action of curcumin thereon.
Asunto(s)
Curcumina/administración & dosificación , Endometriosis/enzimología , Endometriosis/prevención & control , Metaloproteinasa 2 de la Matriz/metabolismo , Enfermedades Peritoneales/enzimología , Enfermedades Peritoneales/prevención & control , Animales , Antiinflamatorios no Esteroideos , Relación Dosis-Respuesta a Droga , Activación Enzimática/efectos de los fármacos , Femenino , Inhibidores de la Metaloproteinasa de la Matriz/administración & dosificación , Ratones , Ratones Endogámicos BALB CRESUMEN
To examine the changes in matrix metalloproteinase-2 (MMP-2) and -9 (MMP-9) in the development and progression of endometriosis, real time quantitative polymerase chain reaction, enzyme-linked immunoabsorbent assay and gelatin zymography were employed to determine the mRNA and protein levels and activities of MMP-2 and MMP-9 from the first day to the 21(st) day after the induction in mice with induced endometriosis (experimental group) and sham-operated animals (controls). The results showed that the mRNA and protein levels and activities of the MMP-2 and MMP-9 were significantly increased on the first day after the induction and the level of MMP-2 stayed at a level higher than that in the control group. MMP-9 had two or three peaks during the 21 days, taking place at day 1, 4 and 15. It is concluded that the changes in the MMP-2 and MMP-9 might be involved in pathogenesis of endometriosis.
Asunto(s)
Endometriosis/enzimología , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Enfermedades Peritoneales/enzimología , Animales , Endometriosis/etiología , Femenino , Metaloproteinasa 2 de la Matriz/genética , Metaloproteinasa 9 de la Matriz/genética , Ratones , Ratones Endogámicos BALB C , Enfermedades Peritoneales/etiología , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
Matrix metalloproteinases (MMPs) modulate extracellular matrix turnover, inflammation and immunity. We studied MMP-9 and MMP-2 in experimental paracoccidioidomycosis. At 15 and 120 days after infection (DAI) with virulent Paracoccidioides brasiliensis, MMP-9 was positive by immunohistochemistry in multinucleated giant cells, in mononuclear cells with macrophage and lymphocyte morphologies and also in fungal cells in the lesions of susceptible and resistant mice. Using gelatin zymography, pro- and active MMP-9 and active MMP-2 were detected in all infected mice, but not in controls. Gelatinolytic activity was not observed in P. brasiliensis extracts. Semiquantitative analysis of gelatinolytic activities revealed weak or absent MMP-2 and strong MMP-9 activity in both mouse strains at 15 DAI, declining at 120 DAI. Avirulent P. brasiliensis-infected mice had residual lesions with MMP-9-positive pseudoxantomatous macrophages, but no gelatinase activity at 120 DAI. Our findings demonstrate the induction of MMPs, particularly MMP-9, in experimental paracoccidioidomycosis, suggesting a possible influence in the pattern of granulomas and in fungal dissemination.
Asunto(s)
Metaloproteinasas de la Matriz/metabolismo , Paracoccidioides , Paracoccidioidomicosis/enzimología , Animales , Femenino , Gelatina/metabolismo , Granuloma/enzimología , Granuloma/microbiología , Técnicas para Inmunoenzimas , Metaloproteinasa 2 de la Matriz/metabolismo , Metaloproteinasa 9 de la Matriz/metabolismo , Ratones , Ratones Endogámicos , Epiplón/enzimología , Enfermedades Peritoneales/enzimología , Enfermedades Peritoneales/microbiologíaRESUMEN
OBJECTIVE: To determine the mechanism by which hypoxia increases expression of iNOS in human normal peritoneal and adhesion fibroblasts. DESIGN: Prospective experimental study. SETTING: University medical center. PATIENT(S): Primary cultures of fibroblasts from normal peritoneum and adhesion tissues. INTERVENTION(S): Hypoxia-treated cells. MAIN OUTCOME MEASURE(S): We used real-time reverse transcription-polymerase chain reaction to quantify mRNA levels of iNOS and nuclear factor kappa B (NF-kappaB). Western blots were used to determine iNOS, NF-kappaB, IkappaB-alpha, and phospho-IkappaB expression levels in normal peritoneal and adhesion fibroblasts in response to hypoxia. RESULT(S): Hypoxia resulted in a significant increase in iNOS and NF-kappaB expression in normal and adhesion fibroblasts. Furthermore, both cell types manifested lower levels of NF-kappaB, cytoplasmic phospho-IkappaB-alpha, and iNOS proteins. In contrast, they manifested higher levels of cytoplasmic IkappaB-alpha and IkappaB-alpha/NF-kappaB ratios as well as a phosphorylated-IkappaB-alpha/NF-kappaB ratio. Under hypoxic conditions, both cell types exhibited significantly decreased cytoplasmic NF-kappaB, IkappaB-alpha levels, and significantly increased cytoplasmic phospho-IkappaB-alpha, iNOS, and NF-kappaB protein levels. CONCLUSION(S): Hypoxia increases iNOS expression by a mechanism involving activation of NF-kappaB. The ratio of IkappaB-alpha/NF-kappaB or IkappaB-alpha/p-IkappaB-alpha can be used to monitor activation.
Asunto(s)
FN-kappa B/metabolismo , Óxido Nítrico Sintasa de Tipo II/metabolismo , Enfermedades Peritoneales/enzimología , Peritoneo/enzimología , Transducción de Señal , Western Blotting , Hipoxia de la Célula , Células Cultivadas , Humanos , Proteínas I-kappa B/metabolismo , Inhibidor NF-kappaB alfa , FN-kappa B/genética , Óxido Nítrico Sintasa de Tipo II/genética , Enfermedades Peritoneales/patología , Peritoneo/patología , Fosforilación , Estudios Prospectivos , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de Tiempo , Adherencias Tisulares , Regulación hacia ArribaRESUMEN
Oxidative stress has been implicated in intra-abdominal adhesion formation. Substance P, a neurokinin-1 receptor (NK-1R) ligand, facilitates leukocyte recruitment and reactive oxygen species (ROS) generation. We have shown in a rat model of adhesion formation that intraperitoneal administration of a NK-1R antagonist at the time of abdominal operation reduces postoperative adhesion formation. Thus we determined the effects of NK-1R antagonist administration on peritoneal leukocyte recruitment and oxidative stress within 24 h of surgery. Adhesions were induced in Wistar rats randomly assigned to receive the antagonist or vehicle intraperitoneally. Peritoneal tissue was isolated at 2, 4, 6, and 24 h after surgery for analysis of the oxidative stress biomarkers 8-isoprostane (8-IP), protein carbonyl, NADPH oxidase, myeloperoxidase (MPO), and ICAM-1 and VCAM-1 mRNAs. Total antioxidant capacity of peritoneal fluid was also determined. MPO, NADPH oxidase, 8-IP, and protein carbonyl were elevated (P < 0.05) by 6 h. ICAM-1 mRNA was elevated (P < 0.05) by 2 h, whereas VCAM-1 levels decreased (P < 0.05) at 24 h. The NK-1R antagonist delayed the MPO rise and reduced (P < 0.05) 8-IP levels by 6 h and ICAM-1 mRNA, VCAM-1 mRNA, and protein carbonyl at 2 h. The antagonist also increased (P < 0.05) the antioxidant capacity of peritoneal fluid at all time points. These data further support a role for oxidative stress in adhesion formation and suggest that the NK-1R antagonist may limit adhesions, in part, by reducing postoperative oxidative stress through an inhibition of neutrophil recruitment and an increase in peritoneal fluid antioxidant capacity.
Asunto(s)
Compuestos Bicíclicos Heterocíclicos con Puentes/farmacología , Fármacos Gastrointestinales/farmacología , Antagonistas del Receptor de Neuroquinina-1 , Infiltración Neutrófila/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Enfermedades Peritoneales/prevención & control , Peritoneo/efectos de los fármacos , Animales , Antioxidantes/metabolismo , Compuestos Bicíclicos Heterocíclicos con Puentes/uso terapéutico , Dinoprost/análogos & derivados , Dinoprost/metabolismo , Modelos Animales de Enfermedad , Fármacos Gastrointestinales/uso terapéutico , Molécula 1 de Adhesión Intercelular/genética , Molécula 1 de Adhesión Intercelular/metabolismo , Laparotomía , Masculino , NADPH Oxidasas/metabolismo , Enfermedades Peritoneales/enzimología , Enfermedades Peritoneales/metabolismo , Peritoneo/enzimología , Peritoneo/metabolismo , Peritoneo/cirugía , Peroxidasa/metabolismo , Carbonilación Proteica/efectos de los fármacos , ARN Mensajero/metabolismo , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismo , Receptores de Neuroquinina-1/metabolismo , Sustancia P/metabolismo , Factores de Tiempo , Adherencias Tisulares/prevención & control , Molécula 1 de Adhesión Celular Vascular/genética , Molécula 1 de Adhesión Celular Vascular/metabolismoRESUMEN
BACKGROUND: Long-term peritoneal dialysis (PD) leads to peritoneal injury. At worst, peritoneal injury leads to encapsulating peritoneal sclerosis (EPS), which is a serious complication of PD. The mortality rate of EPS is extremely high. To perform PD safely, monitoring of peritoneal injury that leads to EPS is a necessity. METHODS: A total of 444 PD patients with end-stage renal disease at 60 centres in Japan were analysed (sex, 54% males; median age, 56 years; median PD duration, 55 months). Matrix metalloproteinase (MMP)-2 and MMP-9 in the peritoneal effluents were analysed with gelatin zymography or enzyme-linked immunosorbent assay. Cells expressing MMP-2 in the peritoneal tissue were investigated immunohistologically with anti-MMP-2 antibodies. Peritoneal solute transport was assessed with the peritoneal equilibration test (PET). RESULTS: The MMP-2 levels in peritoneal effluents obtained with the PET were significantly correlated with the D/P Cr ratio (R = 0.69, P < 0.001) and the D/D0 glucose ratio (R = -0.59, P < 0.001). The MMP-2 levels in patients with mild peritoneal injury, moderate peritoneal injury, severe peritoneal injury (EPS) and infectious peritonitis were significantly higher than those in control patients (P < 0.001, P < 0.001, P < 0.01 and P < 0.05, respectively). MMP-2 was produced by myofibroblast-like mesenchymal cells and macrophages in the peritoneum. The peritoneal effluents from patients with infectious peritonitis showed strong MMP-9 signals. CONCLUSIONS: From these results, MMP-2 levels in peritoneal effluents reflect peritoneal solute transport and changes in MMP-2 levels are associated with peritoneal injury that leads to EPS. MMP-2 may be a useful marker of peritoneal injury, increased solute transport or progression to EPS.
Asunto(s)
Líquido Ascítico/enzimología , Soluciones para Diálisis/farmacocinética , Fallo Renal Crónico/terapia , Metaloproteinasa 2 de la Matriz/metabolismo , Diálisis Peritoneal/efectos adversos , Enfermedades Peritoneales/enzimología , Peritoneo/patología , Anciano , Transporte Biológico Activo/fisiología , Progresión de la Enfermedad , Femenino , Estudios de Seguimiento , Humanos , Incidencia , Japón/epidemiología , Fallo Renal Crónico/enzimología , Masculino , Metaloproteinasa 2 de la Matriz/biosíntesis , Persona de Mediana Edad , Enfermedades Peritoneales/epidemiología , Enfermedades Peritoneales/patología , Peritoneo/enzimología , Pronóstico , Estudios Retrospectivos , Esclerosis , Factores de TiempoRESUMEN
OBJECTIVES: Adhesion formation continues to be an important problem in gastrointestinal surgery. In recent years, methylene blue (MB) has been reported to be an effective agent for preventing peritoneal adhesions. However, its effects on the wound healing process are unknown. In the present study, we investigated the effects of MB on the early and late phases of anastomotic wound healing and on adhesion formation. METHODS: We randomly categorized 92 rats into 2 groups in bursting pressure measurements and 50 rats into 3 groups in the adhesion model. We divided the animals into saline-treated (n = 46) or MB-treated (n = 46) groups. Bursting pressures of the anastomoses were measured on postoperative days 3 and 7. In biochemical studies, tissue hydroxyproline levels, total nitrite/nitrate levels and nitric oxide synthase activity were measured on postoperative days 3 and 7. In the adhesion model, we randomly categorized rats into sham (n = 10), saline-treated (n = 20) and MB-treated (n = 20) groups, and the formation of intraperitoneal adhesions was scored on postoperative day 14. We compared the measurement of bursting pressure and biochemical measurements of tissue hydroxyproline levels, total nitrite/nitrate levels and nitric oxide synthase activity. Histopathological findings of specimens were presented. RESULTS: During the early phase of wound healing (postoperative day 3), bursting pressures, tissue hydroxyproline, total nitrite/nitrate levels and nitric oxide synthase activity in the MB-treated group were significantly lower than those of the saline-treated group. On postoperative day 7, there was no significant difference in these parameters between MB and saline-treated groups. In the adhesion model, MB caused a significant reduction in the formation of peritoneal adhesions. CONCLUSION: MB prevents peritoneal adhesions but causes a significant impairment of anastomotic bursting pressure during the early phase of the wound healing process by its transient inhibitory effect on the nitric oxide pathway.
Asunto(s)
Inhibidores Enzimáticos/uso terapéutico , Azul de Metileno/uso terapéutico , Enfermedades Peritoneales/prevención & control , Peritoneo/patología , Cicatrización de Heridas/efectos de los fármacos , Animales , Modelos Animales de Enfermedad , Estudios de Seguimiento , Laparotomía/efectos adversos , Masculino , Óxido Nítrico Sintasa/metabolismo , Enfermedades Peritoneales/enzimología , Enfermedades Peritoneales/patología , Peritoneo/efectos de los fármacos , Complicaciones Posoperatorias , Ratas , Ratas Sprague-Dawley , Adherencias Tisulares/prevención & control , Resultado del TratamientoRESUMEN
BACKGROUND: Leukocyte apoptosis allows safe removal of potentially harmful cells and facilitates resolution of inflammation. We hypothesized that the number of apoptotic cells changes in a disproportionate fashion in parenchymal organs in response to intra-abdominal infection. MATERIALS AND METHODS: The percentage of apoptotic cells in the liver, spleen, lung, and peripheral blood was evaluated following cecal ligation and puncture (CLP) in mice. Tissue myeloperoxidase (MPO) levels were measured as an index of neutrophil extravasation. RESULTS: Liver & spleen MPO continually increased, while lung MPO remained low after CLP. In parallel to the increase in MPO, liver & spleen apoptosis continually increased throughout the 9-day follow-up period, whereas lung apoptosis remained unchanged. CONCLUSIONS: The distribution of apoptotic cells during intraperitoneal infection occurs in an organ specific manner, with significant increases in the spleen and liver. This distribution likely reflected the clearance of apoptotic cells as the inflammatory focus became contained.
Asunto(s)
Apoptosis , Infecciones Bacterianas/microbiología , Infecciones Bacterianas/patología , Enfermedades Peritoneales/microbiología , Enfermedades Peritoneales/patología , Absceso Abdominal/microbiología , Absceso Abdominal/patología , Animales , Infecciones Bacterianas/enzimología , Recuento de Leucocitos , Hígado/patología , Pulmón/patología , Masculino , Ratones , Especificidad de Órganos , Enfermedades Peritoneales/enzimología , Peroxidasa/metabolismo , Bazo/patologíaRESUMEN
We found, by reverse transcription--real time--polymerase chain reaction, that the expression of aromatase (CYP19) in ovarian, peritoneal endometriosis, and deep endometriotic nodules is significantly different, which strengthens the theory of three distinct clinical entities. Compared with peritoneal endometriosis, ovarian endometriosis exhibits an 8-fold higher expression of aromatase, which suggests that aromatase inhibitors may be particularly active in this form of endometriosis.
Asunto(s)
Aromatasa/metabolismo , Endometriosis/enzimología , Enfermedades del Ovario/enzimología , Enfermedades Peritoneales/enzimología , Enfermedades del Recto/enzimología , Enfermedades Vaginales/enzimología , Femenino , Humanos , Especificidad de Órganos , Distribución TisularRESUMEN
OBJECTIVE: To investigate cyclooxygenase (COX-2) expression within different endometriotic lesions and to assess whether these expression patterns correlate with clinical characteristics. DESIGN: Retrospective cross-sectional study. SETTING: University Hospital. PATIENTS: Seventy patients with histologically confirmed exclusively peritoneal (n=20), ovarian (n=19) or deep-infiltrating (n=31) endometriosis and a detailed medical history. INTERVENTION: Immunohistochemical analysis for COX-2 was performed on 108 endometriotic lesions. MEASUREMENTS AND MAIN RESULTS: COX-2 intensity, percentage of stained glandular endometriotic cells, and correlation of COX-2 expression with clinicopathological parameters. Semiquantitative COX-2 expression did not differ between distinct morphological types of endometriosis and showed no association with the menstrual cycle. Patients with peritoneal-only endometriosis suffering from moderate or severe chronic pelvic pain showed significantly more frequent COX-2 overexpression than asymptomatic patients or patients with minimal symptoms. In patients with exclusively ovarian or deep-infiltrating endometriosis no association between COX-2 expression and clinical parameters, such as chronic pelvic pain, dysmenorrhoea, dyspareunia, sterility, lower urinary tract symptoms or gastrointestinal symptoms was observed. CONCLUSION: Peritoneal endometriotic lesions with increased COX-2 expression have a special relevance for the development of chronic, nonmenstruation-associated, pelvic pain in endometriotic patients. These patients may benefit from therapy with COX-2 inhibitors.
Asunto(s)
Ciclooxigenasa 2/biosíntesis , Endometriosis/enzimología , Dolor Pélvico/etiología , Cavidad Peritoneal/patología , Enfermedades Peritoneales/enzimología , Enfermedad Crónica , Estudios Transversales , Ciclooxigenasa 2/análisis , Endometriosis/complicaciones , Femenino , Humanos , Inmunohistoquímica , Enfermedades Peritoneales/complicaciones , Estudios RetrospectivosRESUMEN
In encapsulating peritoneal sclerosis (EPS), matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases are involved in the remodeling of peritoneal tissue. We measured the MMP-2 concentration in the peritoneal effluents of a patient with EPS who discontinued continuous ambulatory peritoneal dialysis (CAPD) therapy because of ultrafiltration failure and/or underdialysis. First, we report a 58-year-old female patient who discontinued CAPD therapy because of underdialysis. Several months after cessation of CAPD, she complained of slightly blood-colored ascites and had an elevated level of C-reactive protein (CRP) in plasma. She was diagnosed as having clinical early-stage EPS. Peritoneal effluents drained from this case, and from 11 patients who discontinued CAPD therapy because of ultrafiltration failure and/or underdialysis, and who underwent peritoneal lavage with 1.5% dextrose peritoneal dialysis fluid for several months, were analyzed by gelatin zymography. MMP-2 concentration was also measured by enzyme-linked immunosorbent assay (ELISA). MMP-2 concentration in peritoneal effluent of this patient was highest compared with that of the other patients. There was some tendency of a positive correlation between MMP-2 concentration per 1 g protein and D/Pcr, and was negative correlation between MMP-2 concentration per 1 g of protein and D/D0 glucose. We concluded that MMP-2 is involved in the peritoneal remodeling of long-term CAPD therapy and the progression of EPS.
Asunto(s)
Líquido Ascítico/enzimología , Metaloproteinasa 2 de la Matriz/metabolismo , Diálisis Peritoneal Ambulatoria Continua/efectos adversos , Enfermedades Peritoneales/enzimología , Enfermedades Peritoneales/etiología , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Persona de Mediana Edad , Concentración Osmolar , Enfermedades Peritoneales/diagnóstico , Enfermedades Peritoneales/patología , Radiografía Abdominal , Esclerosis , Tomografía Computarizada por Rayos X , UltrasonografíaRESUMEN
OBJECTIVE: To evaluate the effect of iNOS on adhesion formation and to assess whether inhibition of iNOS expression affected adhesion formation according to adhesion maturation days. STUDY DESIGN: Forty Wistar Albino rats were subjected to standardized lesion by cecal abrasion and parietal peritoneal defect and were randomly divided into four groups. Group I (control) received no treatment; groups II-IV received N-acetyl-cystein (NAC) 15 mg/100 g per day intramuscularly on days 4-14, 0-14 and 0-3, respectively, after surgery. On the postoperative 14th day adhesion score, tissue iNOS expression, inflammatory cell reaction (ICR) and tissue fibrosis score were determined. RESULTS: Inflammation score of groups I and II was lower than that of groups III and IV (P < 0.05). Adhesion scores and tissue fibrosis of group II were significantly lower than that of the other groups (P < 0.001). CONCLUSION: iNOS inhibition during the first 3 days postoperatively caused a delay in the resolution of inflammatory cell reaction. On the other hand, when inhibited after the first 3 days, adhesion formation and fibrosis were reduced both clinically and histopathologically.
Asunto(s)
Óxido Nítrico Sintasa/fisiología , Enfermedades Peritoneales/prevención & control , Complicaciones Posoperatorias/prevención & control , Acetilcisteína/farmacología , Animales , Modelos Animales de Enfermedad , Inhibidores Enzimáticos/farmacología , Femenino , Fibrosis/prevención & control , Inflamación/prevención & control , Óxido Nítrico Sintasa/antagonistas & inhibidores , Óxido Nítrico Sintasa de Tipo II , Enfermedades Peritoneales/enzimología , Enfermedades Peritoneales/patología , Peritoneo/cirugía , Complicaciones Posoperatorias/enzimología , Complicaciones Posoperatorias/patología , Ratas , Ratas Wistar , Adherencias Tisulares/enzimología , Adherencias Tisulares/patología , Adherencias Tisulares/prevención & controlRESUMEN
Endometriosis is subsequent to the ability of endometrial glands to invade normal tissues. Matrix metalloproteinases (MMPs)--enzymes that mediate normal tissue turnover, including endometrial breakdown during menstruation-appear to be involved in this invasive process. Here, we examined the immunohistochemical expression of MMP-2, MMP-3, MMP-11, tissue inhibitor metalloproteinase (TIMP)-1 and TIMP-2 in endometrium from women with (n=9) or without endometriosis (n=18) in comparison with peritoneal (n=20), ovarian (n=20) and colorectal endometriosis (n=20). Women with endometriosis showed decreased endometrial MMP-2 expression compared with women without endometriosis (mean+/-SD positive cells: 24.3+/-28.3% and 69.3+/-12.1%), together with loss of MMP-3 expression (0 versus 17.5%+/-20.2). MMP-11, TIMP-1 and TIMP-2 expression was similar in the two groups. Endometrial MMP-2, -3 and -11 expression and TIMP-1 and -2 expression were similar in women with endometriosis and in those with peritoneal endometriosis. MMP-2, -3 and -11 expression was higher in colorectal endometriosis than in ovarian and peritoneal endometriosis. TIMP-2 expression was lower in colorectal endometriosis (P=0.0002) and ovarian endometriotic cysts (P=0.003) than in peritoneal endometriosis. TIMP-1 expression did not vary according to the location of endometriotic lesions. These results suggest that MMP-2 and -3 and TIMP-2 may be involved in the pathogenesis of endometriosis. Interestingly, MMP-2 and -3 overexpression was related to the infiltrative nature of endometriotic lesions, with possible sequential expression from peritoneal to colorectal endometriosis.
Asunto(s)
Endometriosis/enzimología , Endometrio/enzimología , Enfermedades Intestinales/enzimología , Metaloproteinasa 2 de la Matriz/análisis , Metaloproteinasa 3 de la Matriz/análisis , Metaloendopeptidasas/análisis , Enfermedades del Ovario/enzimología , Enfermedades Peritoneales/enzimología , Inhibidor Tisular de Metaloproteinasa-1/análisis , Inhibidor Tisular de Metaloproteinasa-2/análisis , Femenino , Humanos , Inmunohistoquímica , Metaloproteinasa 11 de la MatrizAsunto(s)
11-beta-Hidroxiesteroide Deshidrogenasas/metabolismo , Síndrome de Cushing/enzimología , Epiplón , Ensayos Clínicos como Asunto , Modelos Animales de Enfermedad , Femenino , Regulación Enzimológica de la Expresión Génica , Humanos , Técnicas In Vitro , Isoenzimas , Masculino , Epiplón/enzimología , Enfermedades Peritoneales/enzimologíaRESUMEN
Intra-abdominal adhesion formation is a major complication of serosal repair following surgery, ischaemia or infection, leading to conditions such as intestinal obstruction and infertility. It has been proposed that the persistence of fibrin, due to impaired plasminogen activator activity, results in the formation of adhesions between damaged serosal surfaces. This study aimed to assess the role of fibrinolysis in adhesion formation using mice deficient in either of the plasminogen activator proteases, tissue-type plasminogen activator (tPA) or urokinase-type plasminogen activator (uPA). We hypothesize that, following serosal injury, mice with decreased peritoneal fibrinolytic activity will be more susceptible to adhesion formation. Adhesion formation was induced in tPA- and uPA-deficient and wild-type mice following either surgical trauma to the serosa with haemorrhage and acute or chronic intraperitoneal inflammation. Adhesion formation was assessed from 1 to 4 weeks post-injury. Mice deficient in tPA were more susceptible to adhesion formation following both a surgical insult and a chronic inflammatory episode compared with uPA-deficient and wild-type mice. In addition, the time of maximal adhesion formation varied depending on the nature of the initial insult. It is proposed that the persistence of fibrin due to decreased tPA activity following surgery or chronic inflammation plays a major role in peritoneal adhesion formation.
Asunto(s)
Enfermedades Peritoneales/enzimología , Enfermedades Peritoneales/etiología , Activadores Plasminogénicos/metabolismo , Animales , Fibrina/metabolismo , Humanos , Ratones , Ratones Noqueados , Modelos Biológicos , Activadores Plasminogénicos/deficiencia , Activadores Plasminogénicos/genética , Complicaciones Posoperatorias/enzimología , Complicaciones Posoperatorias/etiología , Adherencias Tisulares/enzimología , Adherencias Tisulares/etiologíaRESUMEN
OBJECTIVE: To assess the presence of matrix metalloproteinase (MMP-1) and tissue inhibitor of MMP (TIMP-1) in peritoneal fluid and serum of subjects with and without adhesions. DESIGN: Cross-sectional study. SETTING: Academic research centers. PATIENT(S): Sixty-three patients who underwent abdominal/pelvic surgery. INTERVENTION(S): MMP-1, TIMP-1, and MMP-1-TIMP-1 complex content. MAIN OUTCOME MEASURE(S): ELISA. RESULT(S): Peritoneal fluids (PF) and sera of subjects with and without peritoneal adhesions contain MMP-1, TIMP-1, and MMP-1-TIMP-1 complex at varying levels with 10- to 100-fold higher TIMP-1 than MMP-1. Compared with serum, PF contains a lower level of MMP-1 in subjects with mild adhesions and without adhesions, higher TIMP-1 in subjects with extensive adhesions, and lower MMP-1-TIMP-1 complex in subjects with moderate adhesions. However, the serum and PF content of MMP-1, TIMP-1, and MMP-1-TIMP-1 complex was not statistically different among subjects with or without adhesions, with the exception of TIMP-1 in PF of subjects with extensive adhesions. MMP1-TIMP-1 ratio indicates that a major portion of MMP-1 is in complex with TIMP-1. There was no age- or gender-dependent difference in MMP-1 and TIMP-1 content in serum or PF. CONCLUSION(S): Despite differences in MMP-1 and TIMP-1 levels in serum and PF of subjects with extensive and moderate adhesions, there is no correlation between MMP-1 and TIMP-1, with the exception of higher TIMP-1 in PF of subjects with extensive adhesions.
Asunto(s)
Líquido Ascítico/metabolismo , Metaloproteinasa 1 de la Matriz/metabolismo , Enfermedades Peritoneales/enzimología , Adherencias Tisulares/enzimología , Inhibidor Tisular de Metaloproteinasa-1/metabolismo , Abdomen/cirugía , Adulto , Anciano , Anciano de 80 o más Años , Líquido Ascítico/patología , Estudios Transversales , Femenino , Humanos , Masculino , Metaloproteinasa 1 de la Matriz/biosíntesis , Metaloproteinasa 1 de la Matriz/sangre , Persona de Mediana Edad , Complicaciones Posoperatorias/enzimología , Estadísticas no Paramétricas , Inhibidor Tisular de Metaloproteinasa-1/biosíntesis , Inhibidor Tisular de Metaloproteinasa-1/sangre , Cicatrización de Heridas/fisiologíaRESUMEN
OBJECTIVE: To compare expression of matrix metalloproteinase (MMP-1) and tissue inhibitor of MMP (TIMP-1) in serosal tissue of intraperitoneal organs and adhesions. DESIGN: Prospective and cross-sectional study. SETTING: Academic research centers. PATIENT(S): Patients undergoing abdominal or pelvic surgery. INTERVENTION(S): MMP-1 and TIMP-1 expression. MAIN OUTCOME MEASURE(S): Expression of messenger ribonucleic acid (mRNA) and protein was measured by using quantitative reverse transcription polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay. RESULT(S): Serosal tissue of intraperitoneal organs and adhesions express MMP-1 and TIMP-1 mRNA and protein at levels that are consistently varied with 10- to 10,000-fold and 2- to 10-fold higher TIMP, mRNA and protein, respectively. Parietal peritoneum, fallopian tubes and ovaries express higher MMP-1 mRNA levels compared with uterus and adhesions; the lowest expression is found in small and large bowels, subcutaneous tissue. and omentum. Expression of TIMP-1 mRNA was less variable; the highest level was found in the uterus and the lowest in subcutaneous tissue and small bowels. There was less variability in MMP-1 and TIMP-1 protein content than mRNA expression; ovaries and adhesions contained the highest MMP-1 and TIMP-1 levels, respectively, and peritoneum contained the lowest. The MMP-1 and TIMP-1 content and ratios further indicate limited MMP-1 proteolytic activity. Although tissues from premenopausal women express more MMP-1 and TIMP-1, expression did not differ by sex or age. CONCLUSION(S): Because MMP-1 and TIMP-1 expression varies consistently among the serosal tissues of peritoneal organs and adhesions, and because tissue injury alters their expression, site-specific variations in expression of these substances may predispose a particular organ to develop more adhesions.
Asunto(s)
Metaloproteinasa 1 de la Matriz/biosíntesis , Enfermedades Peritoneales/enzimología , Peritoneo/enzimología , Adherencias Tisulares/enzimología , Inhibidor Tisular de Metaloproteinasa-1/biosíntesis , Abdomen/cirugía , Adulto , Anciano , Anciano de 80 o más Años , Estudios Transversales , ADN Complementario/química , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Masculino , Metaloproteinasa 1 de la Matriz/química , Metaloproteinasa 1 de la Matriz/genética , Persona de Mediana Edad , Enfermedades Peritoneales/patología , Peritoneo/patología , Estudios Prospectivos , ARN/química , ARN/genética , ARN/aislamiento & purificación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Estadísticas no Paramétricas , Adherencias Tisulares/patología , Inhibidor Tisular de Metaloproteinasa-1/química , Inhibidor Tisular de Metaloproteinasa-1/genéticaRESUMEN
OBJECTIVE: To assess the effect of postsurgical exposure of peritoneal cavity to glove powders, Hydrocote, latex proteins, and lipopolysaccharide (LPS) on eicosanoid production in peritoneal fluid and cellular distribution of eicosanoid enzymes in peritoneal wound during healing. DESIGN: Randomised experimental study. SETTING: Institute for Wound Research, USA. ANIMALS: 360 mice randomised into six groups of 60 each. INTERVENTION: Abrasion of peritoneal cavity followed by instillation of 500 microl of sterile phosphate buffered saline (PBS) alone (Control) or containing 100 microg/ml of Biosorb, Keoflo, Hydrocote, 1 mg/ml of latex proteins, or 12.5 microg/ml of LPS. Mice were killed at 1, 2, 4, 7, 14 and 28 days, and the peritoneal washing obtained from each animal and concentration of eicosanoids measured. Tissue were immunostained for cyclooxygenases and 5-lipoxygenase and thromboxane A2 (TXA2) synthetase. RESULTS: Peritoneal fluid from uninjured controls contained 3.9 (0.8), 5.2 (0.3) and 0.2 (0.02) ng/ml of thromboxane B2 (TXB2), prostaglandin E2 (PGE2) and leukotriene B4 (LTB4), respectively. These increased significantly during the first week to 6.3 (0.3), 11.7 (0.8) and 2.6 (0.1) ng/ml, p<0.05, before returning to baseline by day 14. In all the treated groups the values were significantly higher than in controls (p<0.05). Immunoreactive cyclo-oxygenases, 5-lipoxygenase and TXA2 synthetase proteins were present in various cell types in uninjured skin and peritoneum, incisional and peritoneal wounds and adhesion tissues. Staining was more intense at the site of wounds and paralleled eicosanoid concentrations during healing. There was no difference between exposed and unexposed groups. CONCLUSION: The presence of glove powders, latex proteins and LPS in peritoneal cavity cause increased eicosanoid production and aggravate the normal inflammatory reaction to tissue injury. This may contribute to the inflammatory or immune reactions and development of adhesions caused by glove powders.
Asunto(s)
Eicosanoides/biosíntesis , Guantes Quirúrgicos , Peritoneo/efectos de los fármacos , Peritoneo/enzimología , Polvos/farmacología , Cicatrización de Heridas/efectos de los fármacos , Animales , Líquido Ascítico/química , Eicosanoides/análisis , Ensayo de Inmunoadsorción Enzimática , Femenino , Inmunohistoquímica , Ratones , Ratones Endogámicos , Enfermedades Peritoneales/enzimología , Peritoneo/lesiones , Periodo Posoperatorio , Distribución Aleatoria , Factores de Tiempo , Adherencias Tisulares/enzimologíaRESUMEN
OBJECTIVE: To evaluate the role of lysyl oxidase in postsurgical adhesion formation and incision wound repair. STUDY DESIGN: Female New Zealand rabbits underwent a pelvic-peritoneum adhesion-inducing operation under sterile conditions. In brief, the uterine horns were removed from the abdomen and abraded with surgical gauze and a scalpel blade. The horns were then replaced into the abdominal cavity, the incision was sutured, and the animals were allowed to recover. The animals were killed before lesion development and after 2, 4, 8, and 14 days of postsurgical recovery. The abraded uterine horns, abdominal wall incisional wound and a portion of the sidewall peritoneum were then removed. Total RNA was extracted using the guanidinium thiocyanate-phenol-chloroform method. Northern blot analysis was performed with an [alpha-32P]-labeled lysyl oxidase probe. RESULTS: Lysyl oxidase was expressed during abdominal wall incision repair on days 2 and 4 of postsurgical recovery, declining thereafter (days 8 and 14). In contrast, no increase in lysyl oxidase expression was noted in the uterine horns as compared to the control sidewall peritoneum. CONCLUSION: Lysyl oxidase plays a differential role in the early stages of abdominal wall and uterine horn repair.