RESUMO
Vascular smooth muscle cells (VSMC) are the principal cellular component of the blood vessel wall where they exist in a differentiated state to maintain vascular tone. However, VSMC are not terminally differentiated and can be induced to dediffentiate, proliferate, and migrate. In fact, smooth muscle cell migration from the vascular wall into the lumen of the vessel is a central feature of occlusive vascular pathologies including atherosclerosis and intimal hyperplasia. In vitro, in the presence of an extracellular matrix, cultured vascular smooth muscle cells can migrate and invade the underlying gelatinous matrix, form multicellular nodular aggregations, and secrete the glycoprotein clusterin. Nodular cultures appear to mimic some of the properties of differentiated VSMC, in vivo. Here, to test the hypothesis that clusterin functions to modulate the formation of VSMC nodules and to facilitate cell migration a clusterin negative VSMC clone, SM-CLU13AS (Moulson and Millis, 1999, J Cell Physiol 180:355), was transiently transfected with plasmid pRcCMVCLU that contains the full-length porcine clusterin cDNA sequence under control of the CMV promoter. The transiently transfected VSMC culture expressed and secreted clusterin and formed nodules. To determine if clusterin regulates VSMC migration we used modified Boyden chamber assays. Clusterin, at 10 microg/ml, clearly promotes VSMC migration. In addition, a 15 amino acid synthetic peptide, representing amino acids 118-132 [KQTCMKFYARVCRSG] of the mature clusterin polypeptide, inhibits VSMC attachment to gelatinous substrate. Finally, clusterin appears to have a role in regulating endogenous clusterin expression in the clusterin negative clone. These results clearly establish that clusterin has functional role in VSMC nodule formation and support the conclusion that clusterin is a critical component of smooth muscle cell phenotypic modulation.
Assuntos
Adesão Celular/fisiologia , Glicoproteínas/fisiologia , Chaperonas Moleculares/fisiologia , Músculo Liso Vascular/citologia , Músculo Liso Vascular/fisiologia , Sequência de Aminoácidos , Animais , Aorta Torácica/citologia , Aorta Torácica/fisiologia , Adesão Celular/efeitos dos fármacos , Divisão Celular , Movimento Celular/fisiologia , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Células Clonais , Clusterina , Meios de Cultivo Condicionados , Glicoproteínas/química , Glicoproteínas/genética , Camundongos , Chaperonas Moleculares/química , Chaperonas Moleculares/genética , Dados de Sequência Molecular , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/farmacologia , Plasmídeos , Proteínas Recombinantes/metabolismo , Suínos , Transcrição Gênica , TransfecçãoRESUMO
PURPOSE: To describe the corneal abnormalities and to measure different modalities of corneal sensitivity in corneal lattice dystrophy type II (familial amyloidosis, Finnish type, also known as gelsolin-related amyloidosis and originally as Meretoja syndrome). METHODS: Twenty eyes of 20 patients were examined by in vivo confocal microscopy and noncontact gas esthesiometry. RESULTS: Pleomorphism of, and dense deposits between or posterior to, the basal epithelial cells were frequently observed, as well as a reduction of long nerve fiber bundles in the subbasal nerve plexus. The anterior stroma was altered in most cases, with fibrosis and abnormal extracellular matrix. In 15 corneas, thick anterior and midstromal filaments, corresponding to lattice lines, and in 11 corneas, thin undulated structures were observed. The average mechanical sensitivity threshold of 12 subjects was increased, and in the remaining 8 subjects there was no response, even to the highest intensity of stimuli used. Three patients did not respond to CO(2), 11 to heat, and 2 to cold, but those patients who responded had normal thresholds. Patients with more long nerve fiber bundles per confocal microscopic image had better mechanical and cold sensitivity than patients with fewer nerve fiber bundles. CONCLUSIONS: Lattice lines seem to be related to amyloid material and not to corneal nerves. However, the subbasal nerve density appears reduced, which results mainly in a decrease in mechanical and, to a lesser extent, thermal sensitivity. The location of stromal filaments and undulated structures changes with increasing age.
Assuntos
Amiloidose/patologia , Córnea/inervação , Distrofias Hereditárias da Córnea/patologia , Doenças dos Nervos Cranianos/patologia , Nervo Oftálmico/patologia , Transtornos de Sensação/patologia , Adulto , Idoso , Amiloidose/complicações , Amiloidose/genética , Distrofias Hereditárias da Córnea/etiologia , Doenças dos Nervos Cranianos/etiologia , Técnicas de Diagnóstico Oftalmológico , Feminino , Gelsolina/genética , Humanos , Masculino , Microscopia Confocal , Pessoa de Meia-Idade , Fibras Nervosas/patologia , Transtornos de Sensação/etiologiaRESUMO
BACKGROUND: The purpose of this study was to evaluate graft and patient survival in first-time kidney transplant recipients 60 years old or older, and to identify pretransplant risk factors that predict clinical outcome. METHODS: We reviewed the clinical course of 206 recipients, 60 years old or older, of first kidney transplants at the University of Minnesota. Patient and graft survival were compared with 1640 patients aged 18 to 59 transplanted during the same time period. Regression analysis was performed to identify risk factors that predicted a poor outcome. RESULTS: In patients 60 years old or older, graft survival at one and five years was 86 and 60%, and patient survival at one and five years was 90 and 68%, respectively. Graft and patient survival were decreased compared with recipients aged 18 to 59, but were similar when censored for patient death as a cause of graft loss. A pretransplant history of nonskin malignancy and vascular disease and a current smoking history were risk factors for decreased graft and patient survival. To determine the potential impact of screening for low-risk patients, we evaluated graft and patient survival in patients age > or =60 without these risk factors versus those with one or more risk factors. In the absence of risk factors, both graft and patient survival were significantly improved compared with patients with these risk factors and were equivalent to that of patients aged 18 to 59. CONCLUSIONS: Renal transplantation is a safe and effective therapy for the older renal failure patient. In the absence of identified risk factors, graft survival is equivalent to that seen in younger patients.
Assuntos
Transplante de Rim , Adolescente , Adulto , Fatores Etários , Idoso , Feminino , Sobrevivência de Enxerto , Humanos , Transplante de Rim/mortalidade , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Transplante HomólogoRESUMO
BACKGROUND: Proinflammatory cytokines, such as TNFalpha are involved in the pathogenesis of allergic rhinitis and asthma. OBJECTIVE: Our aim was to study the tear fluid TNFalpha levels in atopic patients before and after topical ocular allergen challenge. METHODS: Thirteen patients were first challenged with topical allergen in the left eye and then dilution buffer in the right eye. Tear fluid samples were collected before the challenges and after 5, 30 and 60 min. Clinical symptoms were scored. Tear fluid TNFalpha concentrations were measured using a double-antibody radioimmunoassay. TNFalpha release rates were calculated using the tear fluid flow in the collection capillary as an estimate for the tear secretion rate. RESULTS: The mean baseline TNFalpha concentrations (ng/L) were 1310 (allergen-challenged eye) and 967 (control eye), and release rates (pg/min) 1.81 (allergen-challenged eye) and 1.39 (control eye), respectively. In the allergen-challenged eye, TNFalpha concentrations and release rates were 1479 and 4.30 (5 min), 1367 and 3.20 (30 min) and 1426 and 3.80 (60 min). In the control eye, TNFalpha concentrations and release rates were 746 and 0.67 (5 min), 1001 and 0.92 (30 min) and 1504 and 1.05 (60 min). The release rates of the allergen-challenged eye were significantly increased at all time points after the challenge when compared with the baseline or the control eye. CONCLUSION: Increased TNFalpha release rates suggest that this cytokine is an early mediator of allergy after conjunctival challenge.
Assuntos
Alérgenos/imunologia , Hipersensibilidade/metabolismo , Lágrimas/química , Fator de Necrose Tumoral alfa/análise , Adolescente , Adulto , Túnica Conjuntiva/imunologia , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Kidney dysfunction after ischemia can be improved by either limiting the initial injury or by enhancing the subsequent proliferative repair process. Adenosine triphosphate (ATP) favorably affects kidney function when it is given shortly after ischemia. We tested whether ATP promotes the proliferative repair response. Rats were subjected to occlusion of the left renal artery for 40 minutes and received an infusion of ATP, 12.5 micromol intravenously over 30 minutes, beginning at reperfusion. Control animals received saline solution or the hydroxyl radical scavenger dimethylthiourea (DMTU). Despite comparable functional protection by DMTU and ATP, only ATP specifically increased DNA synthesis (renal incorporation of tritiated thymidine) to an extent greater than that produced by ischemia alone. In other animals, ribonucleic acid was extracted from kidneys for Northern analysis. Expression of the proto-oncogenes c-fos and c-jun was enhanced in ATP-treated animals as compared with controls. Expression of a histone protein gene (H2b) and thymidine kinase was increased by ischemia but was not additionally affected by ATP. In vitro studies of primary cultures of renal proximal tubule epithelial cells confirmed the ability of ATP to stimulate cellular proliferation as a consequence of stimulation of purinergic P2 receptors, possibly of the P2x subclass. In summary, ATP given after ischemia increased new DNA synthesis and augmented expression of genes critical to cellular proliferation. These beneficial effects were not merely a consequence of limiting initial cellular damage, and they suggest a novel mechanism of action for ATP and other purinergic receptor agonists in renal ischemia.
Assuntos
Trifosfato de Adenosina/farmacologia , Divisão Celular/fisiologia , Isquemia/patologia , Rim/irrigação sanguínea , Receptores Purinérgicos/fisiologia , Animais , Northern Blotting , Hipóxia Celular , Células Cultivadas , Replicação do DNA , Expressão Gênica , Isquemia/genética , Rim/patologia , Rim/fisiopatologia , Masculino , Ratos , Ratos Sprague-DawleyRESUMO
Growth and injury represent recurrent and related themes in the study of progressive renal disease. We have previously demonstrated that a prooxidant diet, one deficient in antioxidants, selenium and vitamin E, induces renal enlargement, proteinuria, mild tubulointerstitial disease and diminished glomerular filtration rate (GFR). Our present study represents continued examination of these processes. We demonstrate that these diets increase thymidine incorporation into DNA and net DNA content in renal tissue, and induce expression of the mRNA for the proto-oncogene, c-myc, and the histone, H2b. We localize increased DNA synthesis as occurring mainly in the distal renal tubular epithelium. These deficient kidneys also exhibit interstitial expansion that parallels the pattern of DNA synthesis in that both processes are more prominent in the medulla than in the cortex. mRNAs for collagens I, III and IV in conjunction with transforming growth factor-beta1 (TGF-beta1) are up-regulated in the kidney in rats maintained on the deficient diet. In complementary in vitro studies, the exposure of rat kidney fibroblasts, NRK 49F cells, to noncytolytic doses of hydrogen peroxide, induces collagen III, collagen IV and TGF-beta1 mRNA. Induction of these genes is also observed in mesangial cells so exposed to noncytolytic doses of hydrogen peroxide. A final aspect of our study was the examination of renal generation of hydrogen peroxide and the profile of the hydrogen peroxide-degrading enzymes. Deficient kidneys exhibit increased mitochondrial generation of hydrogen peroxide independent of oxygen consumption but in conjunction with suppression of glutathione peroxidase mRNA and activity. Lipid peroxidation was increased twofold in the cortex and medulla of the deficient kidneys. Surprisingly, catalase activity, measured in the cortex and medulla, and whole kidney catalase mRNA were also reduced in rats maintained on the antioxidant deficient diet, effects that may further compromise the clearance of hydrogen peroxide. These changes in catalase represent an adverse response to this dietary deficiency, and may be relevant to decreased catalase activity described in chronic renal insufficiency. Thus, a chronic prooxidant state, with features that mimic those of clinical uremia, increases DNA synthesis of renal tubular epithelium, induces mRNA expression for collagens I, III and IV in conjunction with the mRNA for the fibrogenic cytokine, TGF-beta1. Oxidants also induce collagen III, collagen IV and TGF-beta1 mRNA in vitro.
Assuntos
Colágeno/genética , Expressão Gênica/fisiologia , Rim/metabolismo , Fator de Crescimento Transformador beta/genética , Animais , Antioxidantes/metabolismo , Northern Blotting , Catalase/metabolismo , DNA/análise , DNA/biossíntese , Expressão Gênica/efeitos dos fármacos , Peróxido de Hidrogênio/farmacologia , Rim/patologia , Peroxidação de Lipídeos/efeitos dos fármacos , Peroxidação de Lipídeos/fisiologia , Masculino , Mitocôndrias/enzimologia , Oxidantes/farmacologia , Oxirredução , Proto-Oncogenes/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Selênio/deficiência , Deficiência de Vitamina E/metabolismoRESUMO
Clusterin, a glycoprotein with potent cellular cohesive properties, is induced in many organs at times of tissue injury or remodeling. After renal infarction, for example, clusterin is localized to tubular epithelial cells in the peri-infarct zone. The purpose of this study was to examine the spatial and temporal expression of cardiac clusterin after myocardial infarction. Sprague-Dawley rats underwent permanent coronary ligation or sham operation. Hearts were harvested at 6 hours and at 2, 14, and 28 days after infarction. Cardiac clusterin expression was examined by immunohistochemistry and in situ hybridization. Left ventricular clusterin staining was evident at 6 hours and 2 days after myocardial infarction, although not at later time periods. Clusterin was localized to peri-infarct zone myocytes and endothelial cells of this region, and local synthesis of clusterin by myocytes was confirmed by in situ hybridization. Clusterin was not present in inflammatory cells or in left ventricular tissue distant from the infarct. The distribution of clusterin was different from the membrane attack complex of complement (C5b-9), with the latter being present diffusely throughout the infarct zone. Although the role of cardiac clusterin is not known, we speculate that clusterin's cohesive properties serve to promote myocyte interactions that are perturbed in the peri-infarct zone after myocardial infarction.
Assuntos
Glicoproteínas/metabolismo , Chaperonas Moleculares , Infarto do Miocárdio/metabolismo , Animais , Clusterina , Complexo de Ataque à Membrana do Sistema Complemento/metabolismo , Técnicas Imunoenzimáticas , Hibridização In Situ , Masculino , RNA Mensageiro/genética , Ratos , Ratos Sprague-Dawley , Fatores de TempoRESUMO
Clusterin is a ubiquitous glycoprotein induced in many organs, including the kidney, at times of tissue injury and/or remodeling. It is speculated in this study that clusterin preserves cell interactions that are otherwise perturbed by renal insults. The purpose of this study was to examine clusterin expression after cisplatin nephrotoxicity, a model characterized by a delayed time course of injury and a well-defined site of that injury (proximal tubule). Sprague-Dawley rats were treated with intravenous cisplatin (6 mg/kg) or vehicle. Serum creatinine concentrations were measured and kidneys harvested at 1, 2, and 5 days. Marked induction of clusterin mRNA was seen only at 5 days, a time when serum creatinine concentration was the highest. Histology of kidney tissue 5 days after cisplatin administration revealed marked tubular necrosis localized to the outer stripe of the outer medulla, a region rich in proximal tubules. Immunohistochemistry and in situ hybridization at 5 days demonstrated clusterin primarily in the inner stripe of the outer medulla. In conclusion, expression of clusterin follows renal injury with cisplatin at a time corresponding to the morphologic evidence of tubular necrosis and cell detachment; quite surprisingly, such expression occurs at a site distant from the primary injury.
Assuntos
Antineoplásicos/toxicidade , Cisplatino/toxicidade , Glicoproteínas/biossíntese , Rim/efeitos dos fármacos , Rim/metabolismo , Chaperonas Moleculares , Animais , Clusterina , Modelos Animais de Doenças , Glicoproteínas/genética , Rim/lesões , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Fatores de TempoRESUMO
The function of clusterin, a heterodimeric glycoprotein markedly induced in renal and other organ injuries, is unclear. Since renal injury is accompanied by alterations in cell attachment, it is possible that clusterin functions to promote cell-cell and cell-substratum interactions. In this study, a single cell suspension of renal epithelial (LLC-PK1) cells was treated with purified human clusterin, resulting in time- and dose-dependent cell aggregation. Electron microscopy of the cell aggregates demonstrated cell junction and lumen formation. To determine the effect of clusterin on cell adhesion, tissue culture plates were coated with clusterin, fibronectin, PBS, or albumin. Clusterin and fibronectin promoted cell adhesion to the same extent. The adhesion to clusterin was dose dependent and specific, as a monoclonal antibody against clusterin inhibited cell adhesion to clusterin but not fibronectin. Perterbations of the cytoskeleton may underlie the alterations in cell attachment which occur in renal injury. Induction of clusterin mRNA was seen after disruption of both microtubules and microfilaments and after inhibition of cell-substratum interactions. In conclusion, clusterin is a potent renal epithelial cell aggregation and adhesion molecule. We speculate that clusterin functions to promote cell-cell and cell-substratum interactions which are perturbed in the setting of renal injury, thereby preserving the integrity of the renal epithelial barrier.
Assuntos
Adesão Celular/efeitos dos fármacos , Agregação Celular/efeitos dos fármacos , Proteínas Inativadoras do Complemento/farmacologia , Glicoproteínas/farmacologia , Chaperonas Moleculares , Animais , Anticorpos Monoclonais/farmacologia , Linhagem Celular , Clusterina , Proteínas Inativadoras do Complemento/isolamento & purificação , Citoesqueleto/efeitos dos fármacos , Citoesqueleto/fisiologia , Citoesqueleto/ultraestrutura , Relação Dose-Resposta a Droga , Células Epiteliais , Epitélio/efeitos dos fármacos , Epitélio/fisiologia , Fibronectinas/farmacologia , Expressão Gênica , Glicoproteínas/biossíntese , Glicoproteínas/isolamento & purificação , Humanos , Junções Intercelulares/efeitos dos fármacos , Junções Intercelulares/fisiologia , Junções Intercelulares/ultraestrutura , Rim , Cinética , Microvilosidades/efeitos dos fármacos , Microvilosidades/fisiologia , Microvilosidades/ultraestrutura , Suínos , Fatores de TempoRESUMO
Clusterin is a heterodimeric glycoprotein produced by a wide array of tissues and found in most biologic fluids. A number of physiologic functions have been proposed for clusterin based on its distribution and in vitro properties. These include complement regulation, lipid transport, sperm maturation, initiation of apoptosis, endocrine secretion, membrane protection, and promotion of cell interactions. A prominent and defining feature of clusterin is its induction in such disease states as glomerulonephritis, polycystic kidney disease, renal tubular injury, neurodegenerative conditions including Alzheimer's disease, atherosclerosis, and myocardial infarction. The expression of clusterin in these states is puzzling, from the specific molecular species and cellular pathways eliciting such expression, to the roles subserved by clusterin once induced. This review will discuss these physiologic and pathophysiologic aspects of clusterin and speculate on its role in disease.
Assuntos
Glicoproteínas/fisiologia , Chaperonas Moleculares , Doenças Cardiovasculares/metabolismo , Fenômenos Fisiológicos Celulares , Clusterina , Proteínas Inativadoras do Complemento , Glicoproteínas/química , Humanos , Nefropatias/metabolismo , Metabolismo dos Lipídeos , Neoplasias/metabolismo , ReproduçãoRESUMO
Phenol II is a cystogenic chemical that rapidly induces renal cysts, which regress after drug withdrawal. Cyst formation in this model parallels changes in the tubular basement membrane. Clusterin is a potent cohesive factor induced in states of tissue remodeling. The purpose of this study was to determine if renal clusterin was increased in the Phenol II model and to define the time course and distribution of its induction. Male Sprague-Dawley rats were given, by daily gavage, Phenol II (1.2 mg/kg per day) or vehicle (control). The kidneys were harvested after 1, 2, or 4 days of Phenol II treatment or 3 or 7 days after drug withdrawal. An increase in immunoreactive clusterin was seen in the kidneys of Phenol II-treated rats but not in controls. The appearance of clusterin followed a time course similar to that for cyst formation, with expression confined to the epithelial lining and intratubular casts of dilated or cystic tubules. After Phenol II withdrawal, renal cysts regressed and clusterin staining disappeared. The development of cysts was associated with an increase in clusterin mRNA that decreased after drug withdrawal. In conclusion, a marked, yet reversible induction of clusterin occurred in chemically induced polycystic kidney disease. The function of clusterin in this setting remains enigmatic.
Assuntos
Cistos/metabolismo , Glicoproteínas/metabolismo , Nefropatias/metabolismo , Chaperonas Moleculares , Animais , Northern Blotting , Clusterina , Cistos/induzido quimicamente , Cistos/patologia , Glicoproteínas/genética , Imuno-Histoquímica , Nefropatias/induzido quimicamente , Nefropatias/patologia , Túbulos Renais/metabolismo , Túbulos Renais/patologia , Masculino , Fenol , Fenóis , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
Pathological and clinical data from 56 patients operated on for craniopharyngioma since 1981 were analyzed to determine the utility of dividing patients with this tumor into distinct clinical groups based on recognized pathological type and to determine the prognostic import of brain invasion. Of the tumors in the 30 adult patients, 66% were adamantinomatous, 28% were squamous papillary, and the remainder were mixed. However, of the tumors in the 26 children, 96% were adamantinomatous and none were pure squamous papillary (P < 0.01). Forty-six percent of the children compared with 17% of the adults had brain invasion (P < 0.01). Brain invasion was present in 37% of the adamantinomatous but in only 13% of the squamous papillary tumors. Seventy-seven percent of the children underwent gross total resection (GTR) compared with 27% of the adults (P < 0.01). Sixty-three percent of the squamous papillary tumors underwent GTR compared with 54% of the adamantinomatous and mixed tumors. Follow-up ranged from 7 to 187 months (mean, 49 mo). After subtotal resection, with or without radiation therapy, 58% of the tumors recurred compared with 17% recurrence after GTR (P < 0.01), with a mean time to recurrence of 34 months. In both tumor histological types, subtotal resection was associated with a higher rate of tumor recurrence compared with gross total resection. Among the subtotally resected craniopharyngiomas, 2 of the 3 (67%) squamous papillary and 11 of the 21 (52%) adamantinomatous and mixed tumors recurred. In contrast, among the totally resected tumors, none of the 5 squamous papillary and only 5 of the 25 (20%) adamantinomatous and mixed tumors recurred. There were no significant differences in Karnofsky performance status score, mortality rate, or visual and endocrine outcomes when comparing patients based on histological tumor type. When controlling for age and extent of resection, we found that brain invasion had no significant effect on recurrence rate in totally resected tumors. Based on the limited number of patients in this series, we conclude as follows. 1) Contrary to previous reports, squamous papillary craniopharyngiomas, like adamantinomatous tumors, may recur when subtotally resected. 2) For both tumor variants, the most significant factor associated with craniopharyngioma recurrence is the extent of surgical resection rather than histopathological subtype. 3) Contrary to prior hypotheses, brain invasion in totally resected tumors does not predict higher recurrence. 4) GTR is associated with a significantly lower recurrence rate and can be achieved without sacrificing functional outcome.
Assuntos
Craniofaringioma/patologia , Neoplasias Hipofisárias/patologia , Adolescente , Adulto , Idoso , Encéfalo/patologia , Criança , Pré-Escolar , Terapia Combinada , Craniofaringioma/radioterapia , Craniofaringioma/cirurgia , Feminino , Humanos , Hipofisectomia , Lactente , Masculino , Pessoa de Meia-Idade , Invasividade Neoplásica , Recidiva Local de Neoplasia/patologia , Recidiva Local de Neoplasia/radioterapia , Recidiva Local de Neoplasia/cirurgia , Neoplasia Residual/patologia , Neoplasia Residual/radioterapia , Neoplasia Residual/cirurgia , Hipófise/patologia , Irradiação Hipofisária , Neoplasias Hipofisárias/radioterapia , Neoplasias Hipofisárias/cirurgia , Radioterapia AdjuvanteRESUMO
Clusterin is a widely distributed glycoprotein with a wide range of biologic properties. A prominent and defining feature of clusterin is its marked induction in such disease states as glomerulonephritis, cystic renal disease, renal tubular injury, neurodegenerative conditions, atherosclerosis, and myocardial infarction. The expression of clusterin in these states is intriguing given the apparent incongruity of the conditions listed, the variety of stimuli eliciting such expression, and the multiple proposed roles once induced. This review will outline the conditions associated with clusterin induction and speculate on its role in disease.
Assuntos
Apoptose , Glicoproteínas/fisiologia , Chaperonas Moleculares , Animais , Comunicação Celular , Clusterina , Doença , HumanosAssuntos
Angiotensina II/farmacologia , Expressão Gênica/efeitos dos fármacos , Rim/fisiologia , Circulação Renal/fisiologia , Vasoconstrição , Angiotensina II/análogos & derivados , Animais , Pressão Sanguínea/efeitos dos fármacos , Taxa de Filtração Glomerular/efeitos dos fármacos , Rim/efeitos dos fármacos , Rim/metabolismo , Masculino , Norepinefrina/farmacologia , Proto-Oncogenes , Ratos , Ratos Sprague-Dawley , Resistência Vascular/efeitos dos fármacosRESUMO
Clusterin, a glycoprotein with potent cohesive properties, is induced in a wide variety of acute and chronic experimental renal diseases. The purpose of this study was to examine clusterin expression in human renal diseases. Clusterin immunostaining was examined in nephrectomy specimens from patients with autosomal-dominant polycystic kidney disease (N = 5), autosomal-recessive polycystic kidney disease (N = 3), multilocular cyst of the kidney (N = 2), renal hypoplasia/dysplasia (N = 7), Wilms' tumor (nephroblastoma) (N = 6), renal cell carcinoma (N = 9), and acute and/or chronic renal transplant rejection (N = 15). No clusterin staining was detected in normal renal tissue distant from renal cell carcinomas. Increased expression of clusterin was found in epithelial cells lining cysts in all of the cystic disorders studied. Clusterin expression was found in some immature tubules in hypoplastic/dysplastic kidneys and in tubules of rejected renal allografts, but was not a prominent finding in renal neoplasms, although some renal cell carcinomas expressed clusterin in a focal manner. Common features of clusterin induction included exclusively epithelial production of clusterin in cysts, immature nephrons, and injured tubules, heterogeneity of clusterin expression, with only some tubules and/or cysts in a given area staining for clusterin, and uniform clusterin staining of epithelial cells in a given tubule or cyst in most cases. Based on its cohesive properties, we speculate clusterin functions to maintain cell-cell and cell-substratum interactions which become perturbed in the setting of renal injury and cystic diseases.
Assuntos
Glicoproteínas/metabolismo , Nefropatias/metabolismo , Rim/metabolismo , Chaperonas Moleculares , Adolescente , Adulto , Idoso , Anticorpos Monoclonais , Pré-Escolar , Clusterina , Epitélio/metabolismo , Humanos , Técnicas Imunoenzimáticas , Lactente , Recém-Nascido , Rim/patologia , Nefropatias/patologia , Nefropatias/cirurgia , Pessoa de Meia-Idade , NefrectomiaRESUMO
Angiotensin II (Ang II) has growth promoting effects in a number of cells and tissues in vitro. The purpose of this study was to examine the in vivo effect of Ang II on the renal expression of the early growth response genes c-fos, Egr-1 and c-jun. Adult male rats underwent two basal 30 minute clearance periods during which the normal saline vehicle was infused into the left renal artery. Normal saline, Ang II (50 ng/kg/min), or Ang II plus the Ang II antagonist Sar1 Gly8-angiotensin II (10 micrograms/kg/min) were then selectively infused into the left renal artery for two additional 30 minute periods. MAP was similar during basal and Ang II infusion. Hemodynamic effects (decrease in GFR and RPF and increase in renal vascular resistance) were observed only in the Ang II-infused left kidney allowing the right kidney to serve as a paired control for the effects of anesthesia and surgery. Significant increases in the expression of the early growth response genes Egr-1 and c-fos, but not c-jun, were found in the Ang II-infused left kidney compared to the control right kidney. The simultaneous infusion of Ang II (50 ng/kg/min) and the Ang II antagonist Sar1 Gly8-angiotensin II (10 micrograms/kg/min) blocked the increase in Egr-1 and c-fos expression, demonstrating the specificity of the response to Ang II. To compare the effect of another renal vasoconstrictor on the expression of early growth response genes, norepinephrine (40 ng/kg/min) was infused into the left renal artery.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Angiotensina II/farmacologia , Crescimento/efeitos dos fármacos , Rim/efeitos dos fármacos , Norepinefrina/farmacologia , Angiotensina II/fisiologia , Animais , Expressão Gênica/efeitos dos fármacos , Genes fos/efeitos dos fármacos , Genes jun/efeitos dos fármacos , Taxa de Filtração Glomerular/efeitos dos fármacos , Crescimento/genética , Crescimento/fisiologia , Rim/crescimento & desenvolvimento , Rim/fisiologia , Masculino , Norepinefrina/fisiologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Circulação Renal/efeitos dos fármacosRESUMO
To determine whether expression of the renin-angiotensin system (RAS) is influenced by the degree of renal ablation, male Sprague-Dawley rats underwent uninephrectomy, 1 1/3 nephrectomy, or sham operation. Renin and angiotensinogen messenger RNA (mRNA) were not different among the three groups 2 weeks after surgery. The time course of expression of renin mRNA after 1 1/3 nephrectomy showed no difference versus controls at 2 and 4 weeks and a decrease at 6 weeks after surgical ablation. Because nephrons adjacent to the infarcted area in the 1 1/3 nephrectomy may be hypoperfused and a source of increased renin synthesis, intrarenal distribution of tissue renin content, renin mRNA, and immunostainable renin were examined in separate groups of rats subjected to 1 1/3 nephrectomy. The kidney was divided into two pieces, one containing the scar and scar-adjacent tissue and the other portion the tissue distant from the scar. Tissue renin content, renin mRNA, and immunostainable renin were significantly greater in the scar-adjacent tissue compared with the nonscar tissue. Immunoreactive renin was seen in the juxtaglomerular apparatuses as well as in vascular elements proximal to the juxtaglomerular apparatus and within mesangial cells of some glomeruli of the scar-adjacent tissue. In conclusion, immunostainable renin, tissue renin content, and renin mRNA were increased in scar-adjacent tissue after 1 1/3 nephrectomy. We speculate that this unique scar-associated redistribution of renin may play a pathophysiological role in the progression of renal disease.
Assuntos
Rim/fisiologia , Renina/biossíntese , Análise de Variância , Angiotensina II/biossíntese , Animais , Pressão Sanguínea , Northern Blotting , Cicatriz/metabolismo , Cicatriz/patologia , Eletrocoagulação , Técnicas Imunoenzimáticas , Rim/metabolismo , Masculino , Nefrectomia , Período Pós-Operatório , RNA Mensageiro/biossíntese , Ratos , Ratos Endogâmicos , Fatores de Tempo , Transcrição GênicaRESUMO
Clusterin is a multifunctional protein isolated from a number of tissues in several different species. In a variety of renal diseases, clusterin appears in the glomerulus and tubules in association with the membrane attack complex of complement. It is also transiently expressed after several forms of acute renal injury. In this study, we examined the expression and intrarenal distribution of clusterin following subtotal renal ablation. Male rats were subjected to either 1-1/3 nephrectomy (1-1/3 NX), uninephrectomy (UNX) or sham operation (SHAM). Two weeks after surgery, clusterin mRNA was elevated in the 1-1/3 NX group (1-1/3 NX: 1215 +/- 88; UNX: 208 +/- 11; SHAM: 207 +/- 19 OD units; P less than 0.001). Clusterin mRNA increased between 3 and 24 hours after 1-1/3 NX, plateaued, and remained elevated for at least seven weeks. The increased clusterin mRNA in 1-1/3 NX was localized to the tissue adjacent to the infarctive scar (scar 858 +/- 173 vs. non-scar 98 +/- 27 OD units; P less than 0.001). Clusterin protein followed a similar pattern of localization, being increased in most tubules and some peritubular capillaries in the peri-infarct zone. Only occasional tubules were positive for clusterin in the renal tissue distant from the scar or in the kidneys of sham operated rats. Co-localization of clusterin and C5b-9 was not detected. Evidence for apoptosis was found in the peri-infarct zone but not elsewhere in 1-1/3 NX kidney or in the normal kidney following sham operation. Infarction of 1/3 of the left kidney without contralateral nephrectomy, a maneuver which eliminates the compensatory growth, and uremia seen with 1-1/3 NX still resulted in increased clusterin mRNA in the infarcted left kidney compared to the intact right kidney (LK: 790 +/- 112 vs. RK: 128 +/- 25 OD units; P less than 0.001), although the amount of clusterin mRNA was less than that found following 1-1/3 NX. In conclusion, persistently increased clusterin mRNA and protein was seen in the peri-infarct zone following 1-1/3 NX. This increased expression of clusterin may be playing a role in the ischemia-related apoptosis present in the scar-adjacent tissue.
Assuntos
Glicoproteínas/metabolismo , Rim/metabolismo , Chaperonas Moleculares , Nefrectomia , Animais , Clusterina , Expressão Gênica , Glicoproteínas/genética , Imuno-Histoquímica , Infarto/genética , Rim/anatomia & histologia , Rim/cirurgia , Masculino , Nefrectomia/métodos , Período Pós-Operatório , RNA Mensageiro/metabolismo , Ratos , Ratos Endogâmicos , Circulação Renal , Caracteres Sexuais , Fatores de Tempo , Distribuição TecidualRESUMO
Dietary protein restriction reduces proteinuria and slows the progression of renal failure in a variety of renal diseases in native kidneys. Such beneficial effects may be mediated by the multiple renal effects of dietary protein including those on glomerular capillary hemodynamics and the renin-angiotensin system. The role of dietary protein restriction in the management of chronic renal transplant rejection is, however, unclear. This study was therefore undertaken to examine the effects of dietary protein restriction in patients with chronic rejection. Fourteen patients with biopsy proven chronic rejection, who had been on a self-selected home diet of 1.0 +/- 0.1 g protein/kg/day, were randomly assigned, using a crossover design to two 11-day periods, one on a low protein diet (0.55 g/kg/day) and the other on a high protein diet (2 g/kg/day). The effect of these diets on renal hemodynamics, proteinuria, plasma renin activity, and nutritional status was examined. The low protein diet was associated with a significant improvement in glomerular permselectivity in all patients as evidenced by a significant fall in the fractional clearance of albumin and IgG and reduction in 24-hour urinary excretion of total protein, albumin and IgG without any change in blood pressure, glomerular filtration rate, or renal plasma flow. Compared to the proteinuria at the beginning of each diet, a high protein diet did not increase but a low protein diet significantly decreased the proteinuria. The low protein diet was also associated with a significant reduction in plasma renin activity, suggesting that part of the beneficial effect of protein restriction was related to the suppression of the renin-angiotensin system.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Proteínas Alimentares/administração & dosagem , Rejeição de Enxerto , Transplante de Rim/efeitos adversos , Adulto , Proteínas Sanguíneas/metabolismo , Doença Crônica , Feminino , Taxa de Filtração Glomerular , Humanos , Transplante de Rim/fisiologia , Masculino , Proteinúria/dietoterapia , Proteinúria/etiologia , Renina/sangueRESUMO
Recovery from renal ischemia requires regeneration of damaged tubular epithelium. Previous studies have examined the expression of proto-oncogenes and growth factors after ischemia, but the response of genes coding for structural and functional genes has not been scrutinized. Rats were subjected to 40 minutes of renal artery occlusion and 60 minutes to 96 hours of reperfusion. Total RNA was isolated and mRNA for the structural protein actin, the enzymes superoxide dismutase and renin, the proto-oncogene c-fos, the nuclear protein histone H2b, and the putative marker for cell injury TRPM-2 was quantitated by Northern hybridization. Expression of the proto-oncogene c-fos was seen early but for only short duration. Histone gene expression was not markedly increased until 24 hours after ischemia, but remained increased for several days. Renin mRNA was undetectable one hour after ischemia, but was present in normal amounts at 24 and 48 hours. In contrast, superoxide dismutase mRNA was present in decreased amounts 24, 48, and 96 hours after ischemia. TRPM-2 gene expression was greatly increased 24 to 72 hours after ischemia and began decreasing at 96 hours. This selective sequence of gene expression or repression after renal ischemia might maximize the proliferative repair process. This information will be useful for designing therapies to further enhance recovery from acute renal injury.