ABSTRACT
OBJECTIVE@#To explore the marker genes correlated with the prognosis, progression and clinical diagnosis of hepatocellular carcinoma (HCC) based on bioinformatics methods.@*METHODS@#The TCGA-LIHC, GSE84432, GSE143233 and GSE63898 datasets from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) were analyzed. The differentially expressed genes (DEGs) shared by different disease types were obtained using GEO2R and edge R packages, and Gene Ontology (GO) and Kyoto Gene and Genome Encyclopedia (KEGG) enrichment analyses of the DEGs were performed. The expression levels of these DEGs in normal and cancerous tissues were verified in TCGA-LIHC to identify the upregulated genes in HCC. Survival analysis, receiver-operating characteristic (ROC) curve analysis, and correlation analysis between the key genes and the clinical features of the patients were carried out using the R language. The differential expressions of 15 key genes were verified in clinical samples of HCC and adjacent tissues using RT-qPCR.@*RESULTS@#A total of 118 common DEGs were obtained in the database, and among them two genes, namely ATPase Na +/K + transport subunit beta 3 (ATP1B3) and actin regulator (ENAH), showed increased expressions with disease progression. Survival analysis combined with the TCGA-LIHC dataset suggested that high expressions of ATP1B3 and ENAH were both significantly correlated with a poor prognosis of HCC patients (P < 0.05), and their AUC values were 0.821 and 0.933, respectively. A high expression of ATP1B3 was correlated with T stage, pathological stage and pathological grade of the tumors (P < 0.05), while that of ENAH was associated only with an advanced tumor grade (P < 0.05). The results of RT-qPCR showed that ATP1B3 and ENAH were both significantly upregulated in clinical HCC tissues (P < 0.05).@*CONCLUSION@#ATPIB3 and ENAH are both upregulated in HCC, and their high expressions may serve as biomarkers of progression of liver diseases and a poor prognosis of HCC.
Subject(s)
Humans , Carcinoma, Hepatocellular/pathology , Data Mining , Gene Expression Profiling/methods , Gene Expression Regulation, Neoplastic , Liver Neoplasms/pathology , Microfilament Proteins/metabolism , Sodium-Potassium-Exchanging ATPase/metabolismABSTRACT
Abstract Studies in Salvelinus alpinus, Arctic charr, indicate that it has a low capacity to hyposmorregulatory or adaption to sea in winter periods in Arctic waters. The investigation finds to determinate the rank optimum of salinity to can cultivate this species at Chile. The weight adequate was determined to join on the sea by analysis of gill Na+, K+-ATPase activity, that it was found between the ranks 80-130 g, with 14.5 U/mg. It underwent evaluation of fish growth of 72 g salinities from 0 (control), 18, 25 and 33 g/L (sea water) for 94 days. The results indicate that the largest increases were obtained in brackish water. T18 g/L and T25 g/L achieved growth of 25% and 19% on day 94 and term sampling respectively. It is important to mention that the 8% that survived in seawater introduced percentages growth 16.6% equivalent to brackish water and control. These results suggest that Salvelinus alpinus can grow in seawater, with levels of Na+, K+-ATPase similar to those submitted by Salmo salar with a weight not less than 80 g.
Resumo Estudos em Salvelinus alpinus, Charr Ártico, indicam que tem baixa capacidade de hiposmorregulação ou adaptação ao mar em períodos de inverno nas águas do Ártico. A investigação determina o melhor nível de salinidade para cultivar esta espécie no Chile. Determinou-se o peso adequado para se unir ao mar pela análise da atividade da Na +, K + -ATPase das brânquias, que foi encontrada entre as faixas de 80 a 130 g, com 14,5 U/mg. Foi avaliado o crescimento de 72 g salinidades de 0 (controle), 18, 25 e 33 g/L (água do mar) por 94 dias. Os resultados indicam que os maiores aumentos foram obtidos em água salobra. T18 g/L e T25 g/L alcançaram crescimento de 25% e 19% no dia 94 e amostragem a termo, respectivamente. É importante mencionar que os 8% que sobreviveram na água do mar apresentaram percentuais de crescimento de 16,6% equivalentes a água salobra e controle. Estes resultados sugerem que o Salvelinus alpinus pode crescer em água do mar, com níveis de Na +, K + -ATPase semelhantes aos apresentados por Salmo salar com um peso não inferior a 80 g.
Subject(s)
Animals , Sodium-Potassium-Exchanging ATPase/metabolism , Salinity , Trout/metabolism , Chile , GillsABSTRACT
Testicular cancer seminoma is one of the most common types of cancer among men of reproductive age. Patients with this condition usually present reduced semen quality, even before initiating cancer therapy. However, the underlying mechanisms by which testicular cancer seminoma affects male fertility are largely unknown. The aim of this study was to investigate alterations in the sperm proteome of men with seminoma undergoing sperm banking before starting cancer therapy, in comparison to healthy proven fertile men (control group). A routine semen analysis was conducted before cryopreservation of the samples (n = 15 per group). Men with seminoma showed a decrease in sperm motility (P = 0.019), total motile count (P = 0.001), concentration (P = 0.003), and total sperm count (P = 0.001). Quantitative proteomic analysis identified 393 differentially expressed proteins between the study groups. Ten proteins involved in spermatogenesis, sperm function, binding of sperm to the oocyte, and fertilization were selected for validation by western blot. We confirmed the underexpression of heat shock-related 70 kDa protein 2 (P = 0.041), ubiquinol-cytochrome C reductase core protein 2 (P = 0.026), and testis-specific sodium/potassium-transporting ATPase subunit alpha-4 (P = 0.016), as well as the overexpression of angiotensin I converting enzyme (P = 0.005) in the seminoma group. The altered expression levels of these proteins are associated with spermatogenesis dysfunction, reduced sperm kinematics and motility, failure in capacitation and fertilization. The findings of this study may explain the decrease in the fertilizing ability of men with seminoma before starting cancer therapy.
Subject(s)
Adult , Humans , Male , Acrosin/metabolism , Case-Control Studies , Chaperonin Containing TCP-1/metabolism , Electron Transport Complex III/metabolism , HSP70 Heat-Shock Proteins/metabolism , Peptidyl-Dipeptidase A/metabolism , Proteasome Endopeptidase Complex/metabolism , Proteomics , Semen Analysis , Seminoma/metabolism , Sodium-Potassium-Exchanging ATPase/metabolism , Sperm Count , Sperm Motility , Spermatozoa/metabolism , Testicular Neoplasms/metabolismABSTRACT
With aging the kidney exhibits progressive deterioration, with a decrease in renal function. Most of the filtered Na+ is actively reabsorbed in the proximal tubules through different transporters located in apical membrane. This process is possible because basolateral Na+/K+-ATP-ase generates electrochemical conditions necessary for energetically favorable Na+ transport. The α-subunit is the catalytic domain of Na+/K+-ATP-ase. There are three isoforms of the α/subunit present in rat kidney. The present study was undertaken to examine the expression pattern of rat α-Na+/K+-ATP-ase during senescence. We tested the impact of aging on mRNA expression of α-Na+/K+-ATP-ase in cortex and medulla of aged Wistar rats. We observed a significant expression decrease in mRNA levels and a possible change of isoform in the cortex of aged animals. These expression changes observed for αsubunit could be contributing to affect the renal function in conditions of water and salt stress.
Con el avance de la edad los riñones exhiben un deterioro funcional progresivo con disminución de la función renal. La mayor parte del sodio (Na+) filtrado es reabsorbido activamente en los túbulos proximales a través de diferentes transportadores ubicados en la membrana apical. Este proceso es posible por la existencia de la Na+/K+-ATP-asa basolateral, que genera las condiciones electroquímicas necesarias para que el transporte de Na+ sea energéticamente favorable. La subunidad αde la Na+/K+-ATP-asa es el dominio catalítico de la enzima. Existen tres isoformas de subunidad α, que están presentes en el riñón de la rata. En este trabajo se examinan los patrones de expresión de la α-Na+/K+-ATP-asa durante la senescencia. Se estudió así si el aumento de la edad incidía en la expresión del ARNm de la α-Na+/K+-ATP-asa en corteza y médula renal de ratas Wistar senescentes. Se observó una disminución en la expresión del ARNm de la subunidad αy un posible cambio de isoforma predominante en la corteza de los animales senescentes. Los cambios observados para la expresión de la subunidad αpodrían contribuir a afectar la función renal en condiciones de estrés hídrico y salino.
Subject(s)
Animals , Rats , Aging/metabolism , RNA, Messenger/metabolism , Sodium-Potassium-Exchanging ATPase/metabolism , Kidney Cortex/enzymology , Kidney Medulla/enzymology , Sodium/metabolism , RNA, Messenger/analysis , Base Sequence , Random Allocation , Rats, Wistar , Sodium-Potassium-Exchanging ATPase/analysis , Sodium-Potassium-Exchanging ATPase/geneticsABSTRACT
Na+/K+-ATPase (NKA) is abundantly expressed in the basolateral membrane of epithelial cells, which is necessary for tight junction formation. The tight junction is an urothelial barrier between urine and the underlying bladder. Impairment of tight junctions allows migration of urinary solutes in patients with interstitial cystitis/painful bladder syndrome (IC/PBS). We evaluated NKA expression and activity in bladder samples from patients with IC/PBS. The study group consisted of 85 patients with IC/PBS, and the control group consisted of 20 volunteers. Bladder biopsies were taken from both groups. We determined the expression and distribution of NKA using NKA activity assays, immunoblotting, immunohistochemical staining, and immunofluorescent staining. The protein levels and activity of NKA in the study group were significantly lower than the control group (1.08 ± 0.06 vs. 2.39 ± 0.29 and 0.60 ± 0.04 vs. 1.81 ± 0.18 micromol ADP/mg protein/hour, respectively; P < 0.05). Additionally, immunofluorescent staining for detection of CK7, a marker of the bladder urothelium, predominantly colocalized with NKA in patients in the study group. Our results demonstrated the expression and activity of NKA were decreased in bladder biopsies of patients with IC/PBS. These findings suggest that NKA function is impaired in the bladders from patients with IC/PBS.
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Cystitis, Interstitial/diagnosis , Fluorescent Antibody Technique , Keratin-7/metabolism , Microscopy, Fluorescence , Sodium-Potassium-Exchanging ATPase/metabolism , Urinary Bladder/metabolism , Urothelium/metabolismABSTRACT
Introduction Mucoceles are benign expansive cystic formations, composed of a mucus-secreting epithelium (respiratory or pseudostratified epithelium). Nasolacrimal mucocele occurs in a small proportion of children with nasolacrimal duct obstruction and is characterized by a cystic mass in the medial canthus with dilation of the nasolacrimal duct; although dacryocystoceles are rare in adults, they have been reported in patients with trachoma. Objective Discuss clinical aspects, diagnosis, and therapeutic management of mucocele of nasolacrimal duct based on literature review. Resumed Report The authors report a case of bilateral congenital nasolacrimal duct cysts in a 30-year-old man, identified as a tumor in the topography of both lacrimal sacs since birth without associated symptoms. The patient underwent successive surgical treatments, leading to recurrence of the tumor at the right side and recurrent local infections. Conclusion Endoscopic dacryocystorhinostomy has been increasingly used with good results and success rates similar to the external access. .
Subject(s)
Animals , Humans , Genetic Predisposition to Disease/genetics , Mutation/genetics , Nervous System Diseases/genetics , Sodium-Potassium-Exchanging ATPase/genetics , Databases, Bibliographic/statistics & numerical data , Hemiplegia/genetics , Models, Molecular , Nervous System Diseases/diagnosis , Parkinson Disease/genetics , Sodium-Potassium-Exchanging ATPase/metabolismABSTRACT
Till date knowledge regarding the effects of high dietary magnesium on thyroid gland is incomprehensive though certain epidemiological studies reported development of thyroid gland dysfunctions in people with chronic exposure to hard water (especially with high magnesium) despite sufficient iodine consumption. The present study is to explore the effects of chronic high dietary magnesium exposure on thyroid morphology and functional status. Male adult albino Wistar strain rats were treated with graded doses of magnesium sulphate (MgSO4; 0.5, 1.0 and 1.5 g %) for 60 days and changes in different thyroid parameters were investigated. Significantly stimulated thyroid peroxidase and Na+–K+-ATPase and altered idothyronine 5/- deiodinase type I activities, enhanced serum thyroxine (T4) (both total and free), total triiodothyronine (T3) and thyroid stimulating hormone with decreased free T3 levels and T3/T4 ratio (T3:T4) along with enlargement of thyroid with associated histopathological changes were observed in the treated groups. The results clearly confirm that chronic high dietary magnesium exposure causes potential thyroid disruption as reported in earlier epidemiological studies.
Subject(s)
Animals , Dietary Supplements/adverse effects , Iodide Peroxidase/metabolism , Liver/drug effects , Magnesium/administration & dosage , Magnesium/adverse effects , Male , Rats , Sodium-Potassium-Exchanging ATPase/metabolism , Thyroid Gland/cytology , Thyroid Gland/drug effects , Thyroid Gland/enzymology , Thyrotropin/metabolism , Thyroxine/metabolismABSTRACT
Objectives: Fenproporex is an amphetamine-based anorectic which is rapidly converted into amphetamine in vivo. Na+, K+-ATPase is a membrane-bound enzyme necessary to maintain neuronal excitability. Considering that the effects of fenproporex on brain metabolism are poorly known and that Na+, K+-ATPase is essential for normal brain function, this study sought to evaluate the effect of this drug on Na+, K+-ATPase activity in the hippocampus, hypothalamus, prefrontal cortex, and striatum of young rats. Methods: Young male Wistar rats received a single injection of fenproporex (6.25, 12.5, or 25 mg/kg intraperitoneally) or polysorbate 80 (control group). Two hours after the last injection, the rats were killed by decapitation and the brain was removed for evaluation of Na+, K+-ATPase activity. Results: Fenproporex decreased Na+, K+-ATPase activity in the striatum of young rats at doses of 6.25, 12.5, and 25 mg/kg and increased enzyme activity in the hypothalamus at the same doses. Na+, K+-ATPase activity was not affected in the hippocampus or prefrontal cortex. Conclusion: Fenproporex administration decreased Na+, K+-ATPase activity in the striatum even in low doses. However, in the hypothalamus, Na+, K+-ATPase activity was increased. Changes in this enzyme might be the result of the effects of fenproporex on neuronal excitability. .
Subject(s)
Animals , Male , Amphetamines/administration & dosage , Brain/drug effects , Brain/enzymology , Sodium-Potassium-Exchanging ATPase/metabolism , Injections, Intraperitoneal , Rats, Wistar , Time FactorsABSTRACT
There is growing evidence that ouabain, a cardiotonic steroid may promote growth of cardiac and vascular myocytes, indicating its novel role in cell growth and proliferation, without appreciable inhibition of the sodium pump. The mechanism(s) by which low dose of ouabain produces pulmonary artery smooth muscle cell proliferation, a prerequisite for right ventricular hypertrophy, is currently unknown. Here, we analyzed the effects of low dose of ouabain (10 nM) on increase in [Ca2+]i, m-calpain and protein kinase C (PKC) activities on pulmonary artery smooth muscle cell proliferation and determined their sequential involvement in this scenario. We treated bovine pulmonary artery smooth muscle cells with a low dose of ouabain (10 nM) and determined [Ca2+]i in the cells by fluorometric assay using fura2-AM, m-calpain activity by fluorometric assay using SLLVY-AMC as the substrate, PKC activity using an assay kit and assay of Na+/K+ATPase activity spectrophotometrically. We purified m-calpain and PKCα by standard chromatographic procedure by HPLC and then studied cleavage of the purified PKCα by m-calpain using Western immunoblot method. Subsequently, we performed cell proliferation assay utilizing the redox dye resazunin. We used selective inhibitors of [Ca2+]i (BAPTA-AM), m-calpain (MDL28170), PKCα (Go6976) and determined their involvement in ouabain (10 nM)-mediated smooth muscle cell proliferation. Our results suggested that treatment of bovine pulmonary artery smooth muscle cells with a low dose of ouabain (10 nM) increased [Ca2+]i and subsequently stimulated m-calpain activity and proteolytically activated PKCα in caveolae (signaling microdomain also known as signalosomes) of the cells. Upon activation, PKCα increased the smooth muscle cell proliferation via Go/G1 to S/G2-M phase transition. Thus, [Ca2+]i-mCalpain-PKCα signaling axis plays a crucial role during low dose of ouabain-mediated pulmonary artery smooth muscle cell proliferation.
Subject(s)
Amino Acid Sequence , Animals , Calpain/metabolism , Cattle , Caveolae/drug effects , Caveolae/metabolism , Cell Cycle/drug effects , Cell Proliferation/drug effects , Cell Survival/drug effects , Dose-Response Relationship, Drug , Enzyme Activation/drug effects , Molecular Sequence Data , Myocytes, Smooth Muscle/cytology , Myocytes, Smooth Muscle/drug effects , Myocytes, Smooth Muscle/metabolism , Ouabain/pharmacology , Protein Kinase C-alpha/chemistry , Protein Kinase C-alpha/metabolism , Proteolysis/drug effects , Pulmonary Artery/cytology , Sodium-Potassium-Exchanging ATPase/metabolismABSTRACT
Interplay between the host and human cytomegalovirus (HCMV) has a pivotal role in the outcome of infection. A region (referred to as UL/b’) present in the Toledo strain of HCMV and low passage clinical isolates contains 19 additional genes, which are absent in the highly passaged laboratory strain AD169. Products of the UL/b’ genes may determine the manifestations of HCMV infection in vivo. However, little is known about the host factors, which interact with UL/b’ proteins. This study was conducted to investigate the function of the HCMV UL136 protein. By yeast two-hybrid screening, the β1 subunit of the host Na+/K+-ATPase (ATP1B1) was identified to be a candidate protein, which interacts with the HCMV UL136 protein. The interaction was further evaluated both in vitro by pull-down assay and in vivo by immunofluorescent co-localization. The results showed that the UL136 protein can interact with ATP1B1 in vitro. Co-localization of UL136-EGFP and ATP1B1-DsRed in cell membranes suggests that ATP1B1 was a partner of the UL136 protein. It can be proposed that the HCMV UL136 protein may have important roles in processes such as cell-to-cell spread, and in maintaining cell osmotic pressure and intracellular ion homeostasis during HCMV infection.
Subject(s)
Humans , Cytomegalovirus/chemistry , Protein Interaction Mapping , Sodium-Potassium-Exchanging ATPase/metabolism , Two-Hybrid System Techniques , Viral Envelope Proteins/metabolism , Sequence Analysis, ProteinABSTRACT
Background: Infrared (IR) radiation is becoming more popular in industrial manufacturing processes and in many instruments used for diagnostic and therapeutic application to the human eye. Aim: The present study was designed to investigate the effect of IR radiation on rabbit’s crystalline lens and lens membrane. Materials and Methods: Fifteen New Zealand rabbits were used in the present work. The rabbits were classified into three groups; one of them served as control. The other two groups were exposed to IR radiation for 5 or 10 minutes. Animals from these two irradiated groups were subdivided into two subgroups; one of them was decapitated directly after IR exposure, while the other subgroup was decapitated 1 hour post exposure. IR was delivered from a General Electric Lamp model 250R 50/10, placed 20 cm from the rabbit and aimed at each eye. The activity of Na+-K+ ATPase was measured in the lens membrane. Soluble lens proteins were extracted and the following measurements were carried out: estimation of total soluble protein, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Fourier transform infrared (FTIR) spectroscopy. For comparison between multiple groups, analysis of variance was used with significance level set at P < 0.001. Results: The results indicated a change in the molecular weight of different lens crystalline accompanied with changes in protein backbone structure. These changes increased for the groups exposed to IR for 10 minutes. Moreover, the activity of Na+-K+ ATPase significantly decreased for all groups. Conclusions: The protein of eye lens is very sensitive to IR radiation which is hazardous and may lead to cataract.
Subject(s)
Animals , Down-Regulation , Electrophoresis , Female , Infrared Rays , Lens, Crystalline/enzymology , Lens, Crystalline/radiation effects , Male , Rabbits , Sodium-Potassium-Exchanging ATPase/metabolism , Spectroscopy, Fourier Transform Infrared , Time FactorsABSTRACT
In the present study we investigated the effects of lipoic acid (LA) on acetylcholinesterase (AChE), glutathione peroxidase (GPx) and Na+, K+-ATPase activities in rat hippocampus during seizures. Wistar rats were treated with 0.9 percent saline (i.p., control group), lipoic acid (20 mg/kg, i.p., LA group), pilocarpine (400 mg/kg, i.p., P400 group), and the association of pilocarpine (400 mg/kg, i.p.) plus LA (20 mg/kg, i.p.), 30 min before of administration of P400 (LA plus P400 group). After the treatments all groups were observed for 1 h. In P400 group, there was a significant increase in GPx activity as well as a decrease in AChE and Na+, K+-ATPase activities after seizures. In turn, LA plus P400 abolished the appearance of seizures and reversed the decreased in AChE and Na+, K+-ATPase activities produced by seizures, when compared to the P400 seizing group. The results from the present study demonstrate that preadministration of LA abolished seizure episodes induced by pilocarpine in rat, probably by increasing AChE and Na+, K+-ATPase activities in rat hippocampus.
No presente estudo nós investigamos os efeitos do ácido lipóico (AL) sobre as atividades da acetilcolinesterase (AChE), da glutationa peroxidase (GPx) e da Na+, K+-ATPase no hipocampo de ratos durante crises convulsivas. Ratos Wistar foram tratados com solução salina a 0,9 por cento (i.p., grupo controle), ácido lipóico (20 mg/kg, i.p., grupo AL), pilocarpina (400 mg/kg, i.p., grupo P400), e a associação de AL (20 mg/kg, i.p.) com a pilocarpina (400 mg/kg, i.p.), 30 min antes da administração de pilocarpina (grupo AL + P400). Após os tratamentos todos os grupos foram observados durante 1 h. No grupo P400, houve um aumento significativo na atividade da GPx, assim como uma diminuição das atividades da AChE e Na+, K+-ATPase. Por sua vez, o pré-tratamento com AL aboliu o aparecimento de convulsões e reverteu a diminuição das atividades da AChE e da Na+, K+-ATPase causadas pelas convulsões, quando comparada com o grupo P400 sozinho. Os resultados do estudo demonstram que o pré-tratamento com AL aboliu os episódios de convulsão induzido pela pilocarpina em ratos, provavelmente por meio do aumento das atividades das enzimas AChE e Na+, K+-ATPase no hipocampo de ratos.
Subject(s)
Animals , Male , Rats , Acetylcholinesterase/metabolism , Antioxidants/pharmacology , Glutathione Peroxidase/metabolism , Hippocampus/enzymology , Seizures/enzymology , Sodium-Potassium-Exchanging ATPase/metabolism , Thioctic Acid/pharmacology , Hippocampus/drug effects , Pilocarpine , Rats, Wistar , Seizures/chemically inducedABSTRACT
Myocardial infarction leads to compensatory ventricular remodeling. Disturbances in myocardial contractility depend on the active transport of Ca2+ and Na+, which are regulated by Na+-K+ ATPase. Inappropriate regulation of Na+-K+ ATPase activity leads to excessive loss of K+ and gain of Na+ by the cell. We determined the participation of Na+-K+ ATPase in ventricular performance early and late after myocardial infarction. Wistar rats (8-10 per group) underwent left coronary artery ligation (infarcted, Inf) or sham-operation (Sham). Ventricular performance was measured at 3 and 30 days after surgery using the Langendorff technique. Left ventricular systolic pressure was obtained under different ventricular diastolic pressures and increased extracellular Ca2+ concentrations (Ca2+e) and after low and high ouabain concentrations. The baseline coronary perfusion pressure increased 3 days after myocardial infarction and normalized by 30 days (Sham 3 = 88 ± 6; Inf 3 = 130 ± 9; Inf 30 = 92 ± 7 mmHg; P < 0.05). The inotropic response to Ca2+e and ouabain was reduced at 3 and 30 days after myocardial infarction (Ca2+ = 1.25 mM; Sham 3 = 70 ± 3; Inf 3 = 45 ± 2; Inf 30 = 29 ± 3 mmHg; P < 0.05), while the Frank-Starling mechanism was preserved. At 3 and 30 days after myocardial infarction, ventricular Na+-K+ ATPase activity and contractility were reduced. This Na+-K+ ATPase hypoactivity may modify the Na+, K+ and Ca2+ transport across the sarcolemma resulting in ventricular dysfunction.
Subject(s)
Animals , Male , Rats , Myocardial Contraction/physiology , Myocardial Infarction/physiopathology , Sodium-Potassium-Exchanging ATPase/metabolism , Ventricular Function, Left/physiology , Cardiotonic Agents/pharmacology , Myocardial Contraction/drug effects , Myocardial Infarction/enzymology , Ouabain/pharmacology , Rats, Wistar , Vascular Resistance/drug effects , Vascular Resistance/physiology , Ventricular Function, Left/drug effectsABSTRACT
A protein having inhibitory effect on Na+, K+-ATPase as well as showing arylsulphatase A activity (ASA) was isolated from the cytosolic fraction of goat spermatozoa and characterized biochemically. The molecular mass of the protein was found to be 70 kDa (P70) on 10% SDS-PAGE after 35% ammonium sulphate precipitation, followed by hydroxyapatite column chromatographic separation. The isoelectric point (pI) of the protein was found to be 4.9. The sequencing results of first ten N-terminal amino acid residues of protein showed 100%, 90%, and 80% homology with N-terminal 18-27 amino acid residues of mice, pig and human testicular ASA, respectively. The optimum pH, temperature and incubation time for maximum ASA activity of the protein was 5.5, 37°C and 30 min respectively. The ASA activity of protein and AS from a commercial source was studied with respect to the sensitivity to different metal ions, vanadate, carbonyl compounds and ascorbate. Inhibition of AS activity of P70 by silver nitrate suggested that it was related to ASA. Comparable effects of different polyunsaturated fatty acids (eicosapentaenoic and docosahexaenoic acids) and purified anti P70-antibody on P70 and AS from commercial source were observed. The findings suggested that protein was novel in nature, having both regulatory and catalytic functions and showed similarities with the ASA reported from different sources.
Subject(s)
Acrosome Reaction , Animals , Cerebroside-Sulfatase/chemistry , Cerebroside-Sulfatase/genetics , Cerebroside-Sulfatase/metabolism , Enzyme Inhibitors/metabolism , Epididymis/cytology , Goats , Membrane Proteins/chemistry , Membrane Proteins/genetics , Membrane Proteins/metabolism , Molecular Weight , Sodium-Potassium-Exchanging ATPase/antagonists & inhibitors , Sodium-Potassium-Exchanging ATPase/chemistry , Sodium-Potassium-Exchanging ATPase/genetics , Sodium-Potassium-Exchanging ATPase/metabolism , Sperm Capacitation , Spermatozoa/chemistry , Spermatozoa/cytology , Spermatozoa/metabolismABSTRACT
The present study aimed at investigating the influence of increased dietary calcium on Na(+)-K(+)-ATPase activity in heart and aorta of female Sprague-Dawley rats treated with oral contraceptive (OC) steroids. Rats were grouped as control (CR), OC-treated and OC+calcium-treated. OC-treated and OC+calcium-treated received a combination of OC steriods (ethinyloestradiol and norgestrel; ig). OC+calcium-treated rats were fed with 2.5% calcium diet, while OC-treated and CR groups were fed on 0.9% calcium diet. The activity of Na(+)-K(+)-ATPase in heart and aorta was significantly lower in OC-treated rats than those in the other groups. OC treatment caused significant increase in plasma glucose and significant decrease in plasma K+ as compared to control group. Decrease in Na(+)-K(+)-ATPase activity and plasma K+ was abrogated by increased calcium intake, while increase in plasma glucose was not normalized by calcium supplementation. Plasma levels of Na+, lipid peroxidation index and ascorbic acid were comparable among the three groups. These results showed that OC treatment could lead to impaired activity of cardiac and vascular Na(+)-K(+)-ATPase, possibly due to reduced plasma K+ level and these effects could be abolished by high calcium diet.
Subject(s)
Animals , Aorta, Thoracic/drug effects , Blood Glucose/analysis , Blood Vessels/drug effects , Calcium, Dietary/pharmacology , Contraceptives, Oral/pharmacology , Female , Heart/drug effects , Malondialdehyde/blood , Myocardium/enzymology , Oxidative Stress/drug effects , Potassium/blood , Rats , Rats, Sprague-Dawley , Sodium/blood , Sodium-Potassium-Exchanging ATPase/metabolismABSTRACT
Short term effects of insulin on total brain and branchial Na+K+ ATPase, Ca2+ ATPase and Na+, K+ and Ca2+ ions were investigated in A. testudineus. The increase in brain Ca2+ ATPase after alloxan treatment may account for an increased amount of intracellular calcium required for biochemical events taking place inside the cells. Branchial Na+K+ATPase was significantly stimulated while Ca2+ ATPase significantly inhibited after alloxan treatment. This suggests that alloxan exerts its inhibitory effect on the ATP-driven Ca2+ transport via; its action on the Ca2+ pump protein rather than the membrane permeability to Ca2+. The increased activity of brain Na+K+ ATPase at 3 and 24 hr by insulin to alloxan pretreated fish may account for the stimulated co-transport of glucose and its utilization for energy requirements and the excitatory action on neurons in the brain. The elevated brain Ca2+ ATPase may be due to the role of calcium as a second messenger in hormone action. At 24 hr, the activity of branchial Na+K+ ATPase and Ca2+ ATPase in alloxan pretreated specimens was significantly stimulated by insulin. This may be due to increased synthesis of these enzyme units. Administration of insulin (lU/fish) in normal fish significantly inhibited the activity of brain and branchial Na+K+ ATPase while brain Ca2+ ATPase showed a stimulatory effect at 3 and 24 hr compared to control. Inhibition of total branchial Ca2+ ATPase activity by insulin may be due to increased Ca2+ concentration. Higher plasma glucose level in alloxan treated groups confirms the diabetic effect of alloxan. Insulin reverses this effect. The possible mechanism by which insulin controls Na+K+ ATPase activity appears to be tissue specific. The results seem to be the first report on the effect of insulin on ATPase activity in a teleost. These data are consistent with the hypothesis that insulin performs a role in hydro mineral regulation in freshwater teleosts.
Subject(s)
Alloxan/pharmacology , Animals , Blood Glucose/analysis , Brain/drug effects , Calcium/metabolism , Calcium-Transporting ATPases/metabolism , Fish Proteins/metabolism , Gills/drug effects , Insulin/pharmacology , Ions/metabolism , Perciformes/metabolism , Potassium/metabolism , Sodium/metabolism , Sodium-Potassium-Exchanging ATPase/metabolismABSTRACT
BACKGROUND & OBJECTIVES: The biochemical mechanisms underlying the development of sensitization-induced airway hyperresponsiveness (AHR) in asthma are poorly defined. Alterations in the regulation of intracellular calcium may play an important role in its pathogenesis. We carried out this study to see the effect of sensitization with ovalbumin on membrane ion fluxes and intracellular calcium in a guinea pig model. METHODS: Airway reactivity to inhaled histamine was measured initially and after sensitization with ovalbumin in 28 guineapigs. Intracellular calcium [Ca(2+)]i was measured in tracheal smooth muscle cells and peripheral leukocytes using fluorescent dye FURA 2AM. Calcium and sodium ion influx across the cell membrane was measured in leukocytes. Ouabain-sensitive Rubidium ((86)Rb) influx was measured in tracheal smooth muscles cells. The activities of Na(+), K(+) ATPase and Ca(2+) ATPase were measured in tracheal smooth muscle cells. Lipid peroxides were measured in plasma. RESULTS: Airway responsiveness was significantly (P<0.001) increased after sensitization along with an increase in [Ca2+]i levels in leukocytes and tracheal smooth muscle cells, higher rates of (45)Ca and (22)Na influx in leukocytes and higher (86)Rb influx rates in tracheal smooth muscle cells, and increased levels of lipid peroxides in plasma. INTERPRETATION & CONCLUSION: In guineapig model of asthma sensitization to allergen increased the membrane permeability to calcium and sodium, and intracellular calcium levels. These alterations may play a role in the pathogenesis of airway hyper-responsiveness following sensitization.
Subject(s)
Animals , Bronchial Hyperreactivity/metabolism , Calcium/chemistry , Cell Membrane/metabolism , Fluorescent Dyes/metabolism , Fura-2/analogs & derivatives , Guinea Pigs , Histamine/metabolism , Ions/metabolism , Leukocytes/cytology , Lipid Peroxidation , Male , Myocytes, Smooth Muscle/cytology , Ovalbumin/metabolism , Rubidium Radioisotopes/metabolism , Sodium Radioisotopes/metabolism , Sodium-Potassium-Exchanging ATPase/metabolism , Trachea/cytologyABSTRACT
The effect of chronic phostoxin administration on some tissue ATPases, hematology and tissue histopathology was investigated using a combination of gravimetric, enzymatic, colorimetric and histological procedures in New Zealand White rabbits after 2 weeks administration of 0.8mg phostoxin/kg body weight/day, po. The phostoxin treatment led to significant decreases in Na(+)-K+ ATPase activities in renal, hepatic and cardiac tissues. Similar decreases were obtained in the activities of Ca(2+)-ATPase and Mg(2+)-ATPase in liver. In addition, the phostoxin-toxified rabbits manifested significant decreases in hematocrit, red blood cell count, hemoglobin and platelets. Histological examination of the tissues revealed pronounced degenerative changes in liver, heart and kidney.
Subject(s)
Aluminum Compounds/toxicity , Animals , Ca(2+) Mg(2+)-ATPase/metabolism , Calcium-Transporting ATPases/metabolism , Erythrocyte Count , Heart/drug effects , Kidney/drug effects , Liver/drug effects , Pesticides/toxicity , Phosphines/toxicity , Rabbits , Sodium-Potassium-Exchanging ATPase/metabolism , Water Pollutants, Chemical/toxicityABSTRACT
Vitamin E administration prevented DEHP induced deleterious effects like (i) degenerative changes in the brain and thyroid, (ii) decrease in the activity of neuronal membrane Na+ - K+ ATPase, (iii) decrease in the concentration of insulin, cortisol and TSH, and (iv) the increase in T3 and T4 in female Albino rats. The results suggest use of vitamin E to prevent harmful effects of repeated transfusion of DEHP containing blood as in thalassemia patient. The possibility of using vitamin E to prevent the harmful effects of repeated transfusion of DEHP containing blood, as in thalassemia patients, is discussed.
Subject(s)
Animals , Blood Glucose/metabolism , Blood Preservation/methods , Blood Transfusion/methods , Diethylhexyl Phthalate/pharmacology , Female , Hydrocortisone/metabolism , Insulin/metabolism , Plasticizers/chemistry , Polyvinyl Chloride/chemistry , Rats , Rats, Wistar , Sodium-Potassium-Exchanging ATPase/metabolism , Thyrotropin/metabolism , Vitamin E/therapeutic useABSTRACT
The effects of administration of cortisol, corticosterone, testosterone, progesterone and a synthetic estrogen. diethylstilbestrol (DES) on total brain Na(+)-K+- ATPase were investigated in tilapia, O. mossambicus. Exogenous administration of 0.125 and 0.25 microg/g body weight of glucocorticoids and 0.125, 0.25 and 0.5 microg/g body weight of DES for 5 days significantly stimulated Na+(-) K+ ATPase activity by 14-41% in the brain, while 0.5 microg/g body weight of glucocorticoids did not evoke any response on the activity of the enzyme. Progesterone (0.125 and 0.25 microg/g body weight) administration significantly decreased the enzyme activity by 21-36% and high dose (0.5 microg/g body weight) was ineffective. Testosterone exhibited a biphasic effect on Na(+)-K+ ATPase activity--a low dose stimulated by 14% while middle and high doses inhibited it by 19-24%. The results seem to be the first report on the effect of steroids on brain ATPase activity in a teleost. When 0.25microg/g body weight of actinomycin D or puromycin was administered prior to the treatment of similar doses of hormones, the inhibitors significantly inhibited the effect of the hormones by 24-52%. This clearly shows that the effect of the hormones was sensitive to the action of inhibitors suggesting a possible genomic mode of action under long-term treatment. The results suggest that cortisol, corticosterone and DES may possibly stimulate the co-transport of glucose and excitation of membrane potential while progesterone and testosterone inhibit them in the brain of O. mossambicus by regulating the activity of Na(+)-K+ ATPase.