ABSTRACT
OBJECTIVE@#To study the expression of cathepsin-B and -D in different time point after traumatic brain injury.@*METHODS@#Traumatic brain injury (TBI) model was established on rats, cathepsin-B and cathepsin-D immunofluorescence staining and confocal microscope analysis were performed. Positive cells were counted by confocal microscope and image analysis techniques were used to determine the morphological changes in each group.@*RESULTS@#Immunofluorescence staining results showed that cathepsin-B was activated 1 hour after TBI while cathepsin-D was not activated until 12hour after TBI. Both of them got to their peak during 4 to 8days, and kept a high level of activating 32days after TBI. Cathepsin-B and -D positive cells did not merge with caspase-3 positive cells until 6 h after TBI.@*CONCLUSION@#Cathepsin-B and -D could be the diagnostic markers of TBI and can estimating time course of lateral TBI. They blocked caspase-3 activation at the beginning period after TBI and started to promote cell death with caspase-3 6 h after TBI.
Subject(s)
Animals , Male , Rats , Brain/pathology , Brain Injuries/pathology , Caspase 3/metabolism , Cathepsin B/metabolism , Cathepsin D/metabolism , Disease Models, Animal , Forensic Pathology , Hippocampus/pathology , Immunohistochemistry , Lysosomes , Neurons/metabolism , Rats, Sprague-Dawley , Time FactorsABSTRACT
Effect of repeated (20 days) exposure to picrotoxin (PTX) on rat liver lysosomal function was evaluated by measuring the free and total activities of acid phosphatase, cathepsin D, ribonuclease II (RNAse II) and deoxyribonuclease II (DNAse II). The free activities of the nucleases (both RNAse II and DNAse II) were increased following PTX exposure. The total DNAse II activity was increased by 2.2-fold whereas the total acid phosphatase activity was decreased by 28%. Consequently, the ratios of total activity / free activity were low in the PTX exposed groups, implying loss of membrane integrity. Cathepsin D activity was completely abolished. The results show that repeated exposure to PTX can lead to lysosomal dysfunction in liver.
Subject(s)
Acid Phosphatase/metabolism , Animals , Cathepsin D/metabolism , Convulsants/administration & dosage , Endodeoxyribonucleases/metabolism , Exoribonucleases/metabolism , Injections, Intraperitoneal , Liver/drug effects , Lysosomes/drug effects , Male , Picrotoxin/administration & dosage , RatsABSTRACT
BACKGROUND: Immunodeficiency explains the very high frequency of bacterial infections in patients with chronic renal failure (CRF), which leads to high mortality and morbidity, despite improved therapeutic interventions. Among several factors, the decreased functional capacity of phagocytic leucocytes appears to be responsible for the defective host defence mechanisms against infection in CRF. We evaluated both oxygen-dependent and oxygen-independent microbicidal activity of neutrophils isolated from uraemic patients. METHODS: Forty patients with CRF (20 with mild-to-moderate CRF and 20 with advanced CRF) along with 20 age- and sex-matched healthy controls were studied. The assessment of phagocytic capability, ability to produce superoxide (O2.-) anion and H2O2, myeloperoxidase and granule-specific hydrolytic enzymes such as acid phosphatase, cathepsin D and lysozyme activity of the patient's neutrophils were performed to study their bactericidal activity. RESULTS: The phagocytic index (PI) in the control group was found to be 50.38 (4.58). It was significantly reduced in both mild-to-moderate CRF and advanced CRF, as compared to controls. In mild-to-moderate and advanced CRF patients, O2.- production by resting polymorphonuclear neutrophils (PMN) was low. Also, on stimulation with PMA the O2.- production showed a relative reduction as compared to controls. H2O2 production by resting PMN from CRF patients was unaltered but on stimulation with PMA, the quantum of increase was significantly lower. A marked reduction in the level of intracellular myeloperoxidase activity in PMN was noted in CRF patients. Of the three intracellular lysosomal enzymes assayed, cathepsin D level was increased in the PMN of mild-to-moderate CRF patients; acid phosphatase level was elevated significantly in the PMN of both mild-to-moderate and advanced CRF patients. However, no change in lysozyme levels was observed. CONCLUSION: With increasing severity of uraemia, neutrophils from uraemic patients showed progressive impairment of phagocytic ability. Impairment of oxygen-dependent microbicidal mechanisms was indicated by a decrease in O2.- and H2O2 production. Increased activity of lysosomal enzymes such as cathepsin D and acid phosphatase suggest a state of neutrophil activation in uraemia. It is likely that the immunodeficiency state in uraemics is partly due to reduced bactericidal activity of the neutrophil cell population.
Subject(s)
Adolescent , Adult , Cathepsin D/metabolism , Humans , Hydrogen Peroxide/metabolism , Middle Aged , Neutrophils/physiology , Phagocytosis , Superoxides/metabolism , Uremia/immunologyABSTRACT
Single injection of phenylhydrazine[PH] reduced the number of RBC and haemoglobin content; decreased myeloid; erythroid cell ratio in bone marrow and increased Cathepsin D activity in spleen of rats. Ayurvedic drugs raktavardhak, punarnavasav and navayas louh recovered the number of RBC and haemoglobin content and raised myeloid: erythroid cell ratio and normalised cathepsin D activities by counteracting the action phenyl hydrazine. The results confirm the claims of ayurveda that these drugs possess the potency to cure anaemia through protection of RBCs from haemolysis and simultaneously lowering cathepsin D activities from the spleen.
Subject(s)
Anemia, Hemolytic/blood , Animals , Bone Marrow Cells/drug effects , Cathepsin D/metabolism , Erythrocyte Count , Hemoglobins/metabolism , Male , Medicine, Ayurvedic , Phenylhydrazines/toxicity , Plant Extracts/pharmacology , Rats , Rats, Inbred Strains , Spleen/drug effectsABSTRACT
Carbohydrates are the integral parts of glyco-conjugates and play an important role in cellular functions. 2-Deoxy-D-glucose (2-dGlc) is a sugar analogue of glucose and mannose and is reported to inhibit the lipid-linked saccharide formation involved in N-linked glycosylation of proteins. Administration of 2-dGlc (1 mg/100 g body weight) produced a decrease in the tissue total glycosaminoglycans level. We found that the activity of the enzymes involved in the biosynthesis of precursors of glycosaminoglycans (GAG) decreased, but that of the degrading enzymes increased. Thus, the decreased levels of GAG in tissues in 2-dGlc-administered rats occurs via enhanced degradation as well as decreased synthesis.
Subject(s)
Animals , Arteriosclerosis/etiology , Arylsulfatases/metabolism , Cathepsin D/metabolism , Deoxyglucose/pharmacology , Diet, Atherogenic , Glucuronidase/metabolism , Glutamine-Fructose-6-Phosphate Transaminase (Isomerizing)/metabolism , Glycosaminoglycans/metabolism , Glycosylation/drug effects , Hyaluronoglucosaminidase/metabolism , Hypercholesterolemia/complications , Male , Organ Specificity , Protein Precursors/metabolism , Protein Processing, Post-Translational/drug effects , Rats , Rats, Sprague-Dawley , Uridine Diphosphate Glucose Dehydrogenase/metabolism , beta-N-Acetylhexosaminidases/metabolismABSTRACT
Background. A loss of proteins from maternal tissues during lactation has been demonstrated. Protein loss could be explained by intracellular proteolysis. Methods. Cathepsin D activity was studied in the liver, muscle and mammary gland of lactating and weaned rat dams. Lactation was studied at maximal milk production (L-14) and at the final stage of lactation (L-21). Results. Basal activity (virgin rats) was three times higher in liver and mammary gland than in muscle. At both stages, L-14 and L-21, cathepsin D activity increased in liver (50 percent) as well as in the gland (164 percent), but no change was observed in muscle, when compared with controls. Twenty-four hours after litter separation, enzyme activity in the liver decreased to basal levels, while in the mammary gland cathepsin D activity showed a significant decrease but remained higher than control levels. Conclusion. Our results show that liver exhibits adaptive changes in the catabolism of proteins in response to the increased demands imposed by laction on the maternal organism, and when the stimuli disappear activity returns to basal levels. The high activity in mammary gland indicates fast turnover of structures and biomolecules as an answer to the high synthetic activity this tissue. Activity remained higher in the weaning rats, as a result of the regression process which the mammary gland is undergoing
Subject(s)
Female , Breast , Cathepsin D/metabolism , Energy Metabolism , Liver/enzymology , Lactation/metabolism , Muscle, Skeletal/enzymology , Proteins/metabolismABSTRACT
Diurnal changes of lysosomes including ultrastructural changes of phagosomes and acid phosphatase reactions in phagosomes, as well as diurnal biochemical changes in cathepsin D activity, were studied in the retinal pigment epithelium (RPE) of the rabbit. The rabbit was maintained on a natural light-dark cycle over seven days in fall and was sacrificed at various times during the day and night. The number of lysosomes or phagosomes in the RPE was the highest at 1.5 hours after exposure to sunlight (8:00 AM), and thereafter decreased with time. Three types of phagosomes were observed and acid phosphatase reactions were different in each type of phagosome; the fresh phagosomes were negative or positive, lamellar bodies positive, and dense bodies partially positive. The biochemical activity of cathepsin D was the highest at 8:00 AM, and this was consistent with the time of peak in phagocytic activity in the RPE. This report shows that phagocytic activity in the RPE occurred in the early stage after exposure to sunlight, and that fresh phagosomes were sequentially degraded to lamellar or dense bodies. Cathepsin D activity also increased, and this was consistent with the phagocytic activity in the RPE.
Subject(s)
Animals , Rabbits , Acid Phosphatase/metabolism , Cathepsin D/metabolism , Cell Count , Choroid/metabolism , Circadian Rhythm/physiology , Lysosomes/metabolism , Phagosomes/metabolism , Pigment Epithelium of Eye/metabolismABSTRACT
Two different performed HSA-anti-HSA immune aggregates, insoluble complex at equivalence (IC-E) and soluble complex with 5 times antigen excess (IC-S)-were administered iv in experimental mice to study their interaction with liver cells. Both complexes produced no appreciable change in the levels of liver enzymes like acid phosphatase, cathepsin D and gamma-glutamyl transferase. However, marked reduction in the level of liver pseduocholinesterase (as much as 93%) was recorded in the treated animals under identical conditions of administration of both the complexes. Hepatic uptake studies revealed that within 5 min, maximal sequestration of IC occurred within the liver (10 to 18%) and the blood (70 to 82%) when computed in terms of total injected radioactive IC. After 4 h, radioactivity dropped to 3 per cent in liver and 50-40 per cent in blood. The liver seemed to be incapable of scavenging all the serum complexes at a time. Significant consumption of serum complement occurred, when freshly prepared complexes were administered to the animals, but the reduced complement level showed a tendency to reach normalcy after 2 h. The soluble and equivalence zone IC failed to exhibit identifiable discrimination facets with respect to handling by liver. The complexes IC-E and IC-S also behaved in a similar manner.
Subject(s)
Acid Phosphatase/metabolism , Animals , Antigen-Antibody Complex/metabolism , Cathepsin D/metabolism , Humans , Injections, Intravenous , Liver/enzymology , Mice , Mice, Inbred BALB C , Butyrylcholinesterase/metabolism , gamma-Glutamyltransferase/metabolismSubject(s)
Acid Phosphatase/metabolism , Adenosine Triphosphatases/metabolism , Adrenal Glands/drug effects , Animals , Anti-Inflammatory Agents , Catechols/pharmacology , Cathepsin D/metabolism , Curcumin/pharmacology , Female , Glucuronidase/metabolism , Ibuprofen/pharmacology , Inflammation/metabolism , Male , Prostaglandins/metabolism , Rats , Stimulation, ChemicalABSTRACT
Se estudia el efecto que produce el acetato de hidrocortisona y la prednisolona en ratas de 12 días de nacidas que habían sido previamente inyectadas durante 3 días consecutivos con una dosis de 50 mg/kg de peso. Se observó que el efecto del acetato de hidrocortisona fue más intenso que el de la prednisolona al incrementar la actividad de la sacarasa y la leucina aminopeptidasa; y que el efecto de la prednisolona fue más intenso que el del acetato de hidrocortisona al incrementar la actividad de la fosfatasa alcalina y al disminuir la ß galactosidasa ácida y la catepsina D. Además, ninguna de las dos hormonas provocó cambio en la lactato deshidrogenasa. Se concluye que aunque el efecto de ambas hormonas es diferente desde el punto de vista cuantitativo, la prednisolona es capaz de producir los cambios de los niveles enzimáticos de forma similar a los que provoca el acetato de hidrocortisona