Hemiplegia ; diagnosis ; genetics ; Humans ; Mutation ; Prognosis ; Sodium-Potassium-Exchanging ATPase ; genetics

Epilepsy ; etiology ; Hemiplegia ; complications ; genetics ; Humans ; Infant ; Male ; Mutation ; Sodium-Potassium-Exchanging ATPase ; genetics

The presence of serum in a culture medium makes it impossible to identify whether changed cellular functions are directly caused by a manipulation itself or mediated by a component in serum. Madin Darby canine kidney cells can survive in a serum-free medium for about 48 h. We took this advantage to examine whether low K(+)-induced up-regulation of Na,K-ATPase requires serum. We found that serum was essential for low K(+) to induce an increase in Na,K-ATPase binding sites as quantified by ouabain factor binding assays. In an attempt to identify which component was critical, we screened EGF, IGF1, PGE1 and transferrin to identify which one can replace serum. We discovered that transferrin was the single most important factor that mimicked about 80% to 90% of the effect of serum. Transferrin potentiated the effect of low K(+) on the Na,K-ATPase binding sites in a time- and dose-dependent manner. Furthermore, transferrin was also required for low K(+)-induced increase in alpha(1)-promoter activity, alpha(1)- and beta(1)-subunit protein abundance of the Na,K-ATPase. In the presence of transferrin, low K(+) enhanced cellular uptake of iron approximately by 70%. Inhibition of intracellular iron activity by deferoxamine (30 micromol/L) abrogated the effect of low K(+). We conclude that stimulation of the Na,K-ATPase by low K(+) is critically dependent on transferrin. The effect of transferrin is mediated by increased iron transport.
Animals ; Cell Line ; Culture Media, Serum-Free ; Dogs ; Kidney ; cytology ; Potassium ; metabolism ; pharmacology ; Sodium-Potassium-Exchanging ATPase ; genetics ; metabolism ; Transferrin ; metabolism ; pharmacology

OBJECTIVETo analyze the ATP1A3 mutations in patients with alternating hemiplegia of childhood (AHC) and recognize its value in diagnosing atypical cases.

METHODData of all AHC patients seen at Peking University First Hospital from August 2005 to November 2014 were prospectively collected. Clinical information of the AHC patients and their family members were collected and analyzed. Genomic DNAs were extracted from their peripheral blood. Mutations in ATP1A3 were screened by Sanger sequencing after PCR.

RESULTA total of 78 AHC patients were recruited, including 50 males and 28 females. Only three patients had family history of AHC. The first family case had affected mother with AHC; the second family case was the older one of a monozygotic male twins with AHC but their parents were normal; the third family case had a sister with AHC but their parents were normal. The age of onset ranged from six hours to eight years and six months (median: 4 months). According to the Aicardi's clinical diagnostic criteria, 72 patients were considered as typical AHC cases and the other six patients were considered as atypical AHC cases for their age of onset was older than 18 months. Twenty-seven different missense ATP1A3 mutations were detected in 71 (91.0%, 71/78) patients with AHC, including 66 typical and 5 atypical cases. 11 novel ATP1A3 mutations were first reported. ATP1A3 mutations were identified in the three AHC cases with family history. Parental analysis verified that the ATP1A3 mutation of 63 patients (95.5%, 63/66) were de novo origin except lack of five unavailable maternal or paternal genomic DNA. Mutation D801N was found in 20 cases (28.2%), and E815K in 12 cases (16.9%). In the six atypical AHC patients, ATP1A3 mutations were detected in five of them.

CONCLUSIONATP1A3 was the major causative gene of AHC, and mutations were identified as de novo mostly. ATP1A3 mutations in AHC had mutational hotspot, and the most common mutations were D801N and E815K. ATP1A3 mutation screening is helpful for the genetic and definite diagnosis of the atypical AHC cases.

Child ; Child, Preschool ; DNA Mutational Analysis ; Female ; Hemiplegia ; genetics ; Humans ; Infant ; Infant, Newborn ; Male ; Mutation, Missense ; Sodium-Potassium-Exchanging ATPase ; genetics

OBJECTIVETo assess the binding activity of polypeptide containing human Na+, K+-ATPase alpha1 subunit M1-M2 extracellular segment (HES1 derivative).

METHODSHES1 derivative was synthesized by Fmoc method and purified by high-performance liquid chromatography-mass spectrometry, and its binding activity was identified by radioligand binding assay.

RESULTS3H-ouabain and synthetic HES1 derivative showed some binding activity with the equilibrium dissociation constant (KD) of 24.58 nmol/L, with the the receptor density of 492.43 fmol x mg(-1) pro. and IC50 of 3.078 x 10(-7) mol/L.

CONCLUSIONHES1 derivative can bind to ouabain and has the potential of becoming an effective therapeutic agent.

Binding Sites ; drug effects ; Extracellular Space ; metabolism ; Humans ; Ouabain ; chemistry ; pharmacology ; Peptides ; chemistry ; Protein Binding ; Sodium-Potassium-Exchanging ATPase ; chemistry ; genetics ; metabolism

OBJECTIVECardiotonic steroids (CTS) can bind to Na+, K+ -ATPase to activate complex intracellular signaling cascades regulating the proliferation and apoptosis of cells. The aim of this study was to investigate the effects of ouabain at different concentrations on growth regulation in various kinds of leukemia cell lines and explore the pathogenesis of leukemia, the functions of Na+, K+ -ATPase as a signal transduction conductor and its effects on cell growth.

METHODSUsing the MTT assay, the survival rates of leukemia cell lines were observed 6, 12 and 24 hrs after treatment with 1 or 10 nmol/L ouabain. The expression of Na+, K+ -ATPase alpha1 subunit of leukemia cells was detected by Western blot.

RESULTSThe MTT results showed that ouabain at 1 nmol/L or 10 nmol/L induced proliferation of lymphocytic leukemia B95 and Jhhan cell lines, as well as megakaryocytic leukemia M07e and Meg01 cell lines. Ouabain at 1 nmol/L or 10 nmol/L increased the expression of Na+, K+ -ATPase alpha1 subunit. There were significant differences in the proliferation and the expression of Na+, K+ -ATPase alpha1 subunit of the leukemia cell lines between the ouabain treatment and the blank control groups 24 hrs after ouabain treatment (P<0.05). The proliferation effect of leukemia cell lines was in a direct proportion with the ouabain concentration and incubation time.

CONCLUSIONSNa+, K+ -ATPase plays an important role in signal transductions. Through binding to ouabain, Na+, K+ -ATPase may regulate proliferation of leukemia cell lines of different origins. Ouabain at 1 nmol/L or 10 nmol/L may induce proliferation of lymphocytic leukemia cell lines (B95, Jhhan) and megakaryocytic leukemia cell lines (M07e, Meg01), and the proliferation effect was in a direct proportion with the concentration and incubation time of ouabain.

Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Leukemia ; pathology ; Ouabain ; pharmacology ; Signal Transduction ; Sodium-Potassium-Exchanging ATPase ; analysis ; genetics ; physiology

CAPOS syndrome is an autosomal dominant neurological disorder caused by mutations in the ATP1A3 gene. Initial symptoms, often fever-induced, include recurrent acute ataxic encephalopathy in childhood, featuring cerebellar ataxia, optic atrophy, areflflexia, sensorineural hearing loss, and in some cases, pes cavus. This report details a case of CAPOS syndrome resulting from a maternal ATP1A3 gene mutation. Both the child and her mother exhibited symptoms post-febrile induction,including severe sensorineural hearing loss in both ears, ataxia, areflexia, and decreased vision. Additionally, the patient's mother presented with pes cavus. Genetic testing revealed a c. 2452G>A（Glu818Lys） heterozygous mutation in theATP1A3 gene in the patient . This article aims to enhance clinicians' understanding of CAPOS syndrome, emphasizing the case's clinical characteristics, diagnostic process, treatment, and its correlation with genotypeic findings.
Humans ; Child ; Female ; Cerebellar Ataxia/diagnosis* ; Talipes Cavus ; Hearing Loss, Sensorineural/diagnosis* ; Optic Atrophy/diagnosis* ; Mutation ; Phenotype ; Sodium-Potassium-Exchanging ATPase/genetics* ; Foot Deformities, Congenital ; Reflex, Abnormal

OBJECTIVETo investigate the association of the polymorphisms of rs4961 in alpha-adducin (ADD1) and rs28933400 in Na+/K+ -ATPase a2 (ATP1A2) genes, the products of which are important for sodium transport, with essential hypertension.

METHODSMutagenically separated PCR (MS-PCR) was used to detect the genotypes of the two loci. The subjects were recruited randomly including 196 patients of essential hypertension and 192 healthy controls.

RESULTSThe frequencies of genotypes and alleles of in the ADD1 gene were significantly different between the patients and controls respectively (P=0.03, P=0.04). There was significant relationship between the genotypes of rs4961 and systolic blood pressure and blood sodium concentration. However, there was no significant relationship between the rs4961 genotypes and diastolic blood pressure, body mass index, blood kalium and chlorine concentrations. There was no polymorphism at the rs28933400 locus in the subjects analyzed.

CONCLUSIONThe rs4961 polymorphism of the ADD1 gene is associated with essential hypertension, but the rs28933400 locus in the ATP1A2 gene may have no association with essential hypertension in the studied population.

Calmodulin-Binding Proteins ; genetics ; Case-Control Studies ; Female ; Gene Frequency ; Genotype ; Humans ; Hypertension ; genetics ; pathology ; physiopathology ; Ion Transport ; Male ; Middle Aged ; Polymorphism, Genetic ; Sodium ; metabolism ; Sodium-Potassium-Exchanging ATPase ; genetics

Paeoniflorin is the main active ingredient of Chinese herbaceous peony. This study is to investigate the protective effect of paeoniflorin (Pae) on acute brain damage induced by lipopolysaccharide (LPS) in mice. The mice were randomly assigned to the normal control, model control (LPS), as well as groups of paeoniflorin and lipopolysaccharide (Pae + LPS). Then the mice were administered intraperitioneally with normal saline or Pae (10, 30 mg · kg(-1)) once daily for 6 d. One hour after intrapertioneally treatment on the seventh day, each group were injected LPS (5 mg · kg(-1)) to establish the endotoxin lipopolysaccharide inflammation model except the normal group. The mice were sacrificed after 6 h and the brain homogenates were prepared and measured. The malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), total antioxidant capacity (T-AOC), hydrogen peroxide (H2O2), succinatedehydrogenase (SDH), Na(+)-K(+)-ATPase and Ca(2+)-Mg(2+)-ATPase were dectected by the colorimetric method. The levels of HO-1 and Nrf2 protein in subcellular fractions of brain tissue were detected by Western blot. The results demonstrated that the administration with paeoniflorin reduced the levels of the MDA production; significantly increase the activities of antioxidant enzyme (SOD and GSH-PX). In addition, paeoniflorin could enhance the total antioxidant capacity, decrease the level of H2O2, and increase the activities of SDH, Na(+)-K(+)-ATPase and Ca(2+)-Mg(2+)-ATPase. Furthermore, paeoniflorin can increase the expression of HO-1 and activate the nuclear transfer of Nrf2. Taking together, these findings suggest that paeoniflorin alleviate the acute inflammation in mice brain damage induced by LPS, which is related with its antioxidant effect and improvement of energy metabolism.
Animals ; Energy Metabolism ; drug effects ; Glucosides ; pharmacology ; Heme Oxygenase-1 ; genetics ; Lipopolysaccharides ; pharmacology ; Male ; Membrane Proteins ; genetics ; Mice ; Mice, Inbred BALB C ; Monoterpenes ; pharmacology ; Oxidative Stress ; drug effects ; Sodium-Potassium-Exchanging ATPase ; metabolism

AIMTo investigate the mechanisms of Naoyikang (Traditional Chinese Medicine) on the Alzheimer's Disease (AD) model mice induced by D-galactose (D-gal) and NaNO2.

METHODSThe mouse model was established by intraperitoneal injection of D-gal and NaNO2. The capacity of learning and memory was tested on mice with electrical maze; the content of nitric oxide (NO) and the activity of monoamine oxidase-B (MAO-B), glutathione peroxidase (GSH-PX), Na(+) -K(+) -ATP enzyme and Ca(2+) -ATP enzyme in cerebral cortex and hippocampus were assayed by biochemical methods; expression of Bax and Bcl-2 mRNA was detested by RT-PCR.

RESULTSNaoyikang could ameliorate the capacity of learning and memory of AD model mice and reduce MAO-B activity in the brain tissue and activate the activity of Na(+) -K(+) -ATP enzyme and Ca(2+) -ATP enzyme in the brain tissue and decrease the expression of Bax mRNA, but increase the expression of Bcl-2 mRNA in the model brain tissue.

CONCLUSIONNaoyikang could protect AD model mice induced by D-gal and NaNO2. It could modify the metabolism of monoamine neurotransmitter in brain through reducing MAO-B activity and protect neurons by activating the activity of Na(+) -K(+) -ATP enzyme and Ca(2+) -ATP enzyme and decrease Bax expression and increase Bcl-2 expression in the model brain tissue.

Alzheimer Disease ; chemically induced ; drug therapy ; Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Galactose ; Male ; Maze Learning ; Mice ; Mice, Inbred ICR ; Neuroprotective Agents ; therapeutic use ; Phytotherapy ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Sodium Nitrite ; Sodium-Potassium-Exchanging ATPase ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism