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We are analyzing https://link.springer.com/article/10.1007/s10571-004-1379-6.

Title:
Impaired Regulation of pH Homeostasis by Oxidative Stress in Rat Brain Capillary Endothelial Cells | Cellular and Molecular Neurobiology
Description:
1. Endothelial cells are permanently challenged by altering pH in the blood, and oxidative damage could also influence the intracellular pH (pHi) of the endothelium. Cerebral microvascular endothelial cells form the blood–brain barrier (BBB) and pHi regulation of brain capillary endothelial cells is important for the maintenance of BBB integrity. The aim of this study was to address the pH regulatory mechanisms and the effect of an acute exposure to hydrogen peroxide (H2O2) on the pH regulation in primary rat brain capillary endothelial (RBCE) cells. The RBCE monolayers were loaded with the fluorescent pH indicator BCECF and pHi was monitored by detecting the fluorescent changes. 2. The steady-state pHi of RBCE cells in HEPES-buffer (6.83 ± 0.1) did not differ significantly from that found in bicarbonate-buffered medium (6.90 ± 0.08). Cells were exposed to NH4Cl to induce intracellular acidification and then the recovery to resting pH was studied. Half-recovery time after NH4Cl prepulse-induced acid load was significantly less in the bicarbonate-buffered medium than in the HEPES-medium, suggesting that in addition to the Na+/H+ exchanger, HCO3 −/Cl− exchange mechanism is also involved in the restoration of pHi after an intracellular acid load in primary RBCE cells. We used RT-PCR-reactions to detect the isoforms of Na+/H+ exchanger gene family (NHE). NHE-1 -2, -3 and -4 were equally present, and there was no significant difference in the relative abundance of the four transcripts in these cells. 3. No pHi recovery was detected when the washout after an intracellular acid load occurred in nominally Na+-free HEPES-buffered medium or in the presence of 10 ÎŒM 5-(N-ethyl-N-isopropyl)amiloride (EIPA), a specific inhibitor of Na+/H+ exchanger. The new steady-state pHi were 6.37 ± 0.02 and 6.60 ± 0.02, respectively. 4. No detectable change was observed in the steady-state pHi in the presence of 100 ÎŒM H2O2; however, recovery from NH4Cl prepulse-induced intracellular acid load was inhibited when H2O2 was present in 50 or 100 ÎŒM concentration in the HEPES-buffered medium during NH4Cl washout. These data suggest that H2O2 is without effect on the activity of Na+/H+ exchanger at rest, but could inhibit the function of the exchanger after an intracellular acid load.
Website Age:
28 years and 1 months (reg. 1997-05-29).

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🌠 Phenomenal Traffic: 5M - 10M visitors per month


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Keywords {🔍}

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Topics {✒}

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Schema {đŸ—ș}

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      mainEntity:
         headline:Impaired Regulation of pH Homeostasis by Oxidative Stress in Rat Brain Capillary Endothelial Cells
         description:1. Endothelial cells are permanently challenged by altering pH in the blood, and oxidative damage could also influence the intracellular pH (pHi) of the endothelium. Cerebral microvascular endothelial cells form the blood–brain barrier (BBB) and pHi regulation of brain capillary endothelial cells is important for the maintenance of BBB integrity. The aim of this study was to address the pH regulatory mechanisms and the effect of an acute exposure to hydrogen peroxide (H2O2) on the pH regulation in primary rat brain capillary endothelial (RBCE) cells. The RBCE monolayers were loaded with the fluorescent pH indicator BCECF and pHi was monitored by detecting the fluorescent changes. 2. The steady-state pHi of RBCE cells in HEPES-buffer (6.83 ± 0.1) did not differ significantly from that found in bicarbonate-buffered medium (6.90 ± 0.08). Cells were exposed to NH4Cl to induce intracellular acidification and then the recovery to resting pH was studied. Half-recovery time after NH4Cl prepulse-induced acid load was significantly less in the bicarbonate-buffered medium than in the HEPES-medium, suggesting that in addition to the Na+/H+ exchanger, HCO3 −/Cl− exchange mechanism is also involved in the restoration of pHi after an intracellular acid load in primary RBCE cells. We used RT-PCR-reactions to detect the isoforms of Na+/H+ exchanger gene family (NHE). NHE-1 -2, -3 and -4 were equally present, and there was no significant difference in the relative abundance of the four transcripts in these cells. 3. No pHi recovery was detected when the washout after an intracellular acid load occurred in nominally Na+-free HEPES-buffered medium or in the presence of 10 ÎŒM 5-(N-ethyl-N-isopropyl)amiloride (EIPA), a specific inhibitor of Na+/H+ exchanger. The new steady-state pHi were 6.37 ± 0.02 and 6.60 ± 0.02, respectively. 4. No detectable change was observed in the steady-state pHi in the presence of 100 ÎŒM H2O2; however, recovery from NH4Cl prepulse-induced intracellular acid load was inhibited when H2O2 was present in 50 or 100 ÎŒM concentration in the HEPES-buffered medium during NH4Cl washout. These data suggest that H2O2 is without effect on the activity of Na+/H+ exchanger at rest, but could inhibit the function of the exchanger after an intracellular acid load.
         datePublished:
         dateModified:
         pageStart:141
         pageEnd:151
         sameAs:https://doi.org/10.1007/s10571-004-1379-6
         keywords:
            blood–brain barrier
            intracellular pH
            rat brain capillary endothelial cells
            BCECF
            hydrogen peroxide
            sodium/hydrogen exchange
            Neurosciences
            Cell Biology
            Neurobiology
         image:
         isPartOf:
            name:Cellular and Molecular Neurobiology
            issn:
               1573-6830
               0272-4340
            volumeNumber:25
            type:
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               PublicationVolume
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            name:Kluwer Academic Publishers-Plenum Publishers
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         author:
               name:IldikĂł Sipos
               affiliation:
                     name:Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences
                     address:
                        name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences, Hungary
                        type:PostalAddress
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               name:BeĂĄta Tör’ócsik
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                     name:Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences
                     address:
                        name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences, Hungary
                        type:PostalAddress
                     type:Organization
               type:Person
               name:Laszlo Tretter
               affiliation:
                     name:Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences
                     address:
                        name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences, Hungary
                        type:PostalAddress
                     type:Organization
               type:Person
               name:Vera Adam-Vizi
               affiliation:
                     name:Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences
                     address:
                        name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences, Hungary
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      headline:Impaired Regulation of pH Homeostasis by Oxidative Stress in Rat Brain Capillary Endothelial Cells
      description:1. Endothelial cells are permanently challenged by altering pH in the blood, and oxidative damage could also influence the intracellular pH (pHi) of the endothelium. Cerebral microvascular endothelial cells form the blood–brain barrier (BBB) and pHi regulation of brain capillary endothelial cells is important for the maintenance of BBB integrity. The aim of this study was to address the pH regulatory mechanisms and the effect of an acute exposure to hydrogen peroxide (H2O2) on the pH regulation in primary rat brain capillary endothelial (RBCE) cells. The RBCE monolayers were loaded with the fluorescent pH indicator BCECF and pHi was monitored by detecting the fluorescent changes. 2. The steady-state pHi of RBCE cells in HEPES-buffer (6.83 ± 0.1) did not differ significantly from that found in bicarbonate-buffered medium (6.90 ± 0.08). Cells were exposed to NH4Cl to induce intracellular acidification and then the recovery to resting pH was studied. Half-recovery time after NH4Cl prepulse-induced acid load was significantly less in the bicarbonate-buffered medium than in the HEPES-medium, suggesting that in addition to the Na+/H+ exchanger, HCO3 −/Cl− exchange mechanism is also involved in the restoration of pHi after an intracellular acid load in primary RBCE cells. We used RT-PCR-reactions to detect the isoforms of Na+/H+ exchanger gene family (NHE). NHE-1 -2, -3 and -4 were equally present, and there was no significant difference in the relative abundance of the four transcripts in these cells. 3. No pHi recovery was detected when the washout after an intracellular acid load occurred in nominally Na+-free HEPES-buffered medium or in the presence of 10 ÎŒM 5-(N-ethyl-N-isopropyl)amiloride (EIPA), a specific inhibitor of Na+/H+ exchanger. The new steady-state pHi were 6.37 ± 0.02 and 6.60 ± 0.02, respectively. 4. No detectable change was observed in the steady-state pHi in the presence of 100 ÎŒM H2O2; however, recovery from NH4Cl prepulse-induced intracellular acid load was inhibited when H2O2 was present in 50 or 100 ÎŒM concentration in the HEPES-buffered medium during NH4Cl washout. These data suggest that H2O2 is without effect on the activity of Na+/H+ exchanger at rest, but could inhibit the function of the exchanger after an intracellular acid load.
      datePublished:
      dateModified:
      pageStart:141
      pageEnd:151
      sameAs:https://doi.org/10.1007/s10571-004-1379-6
      keywords:
         blood–brain barrier
         intracellular pH
         rat brain capillary endothelial cells
         BCECF
         hydrogen peroxide
         sodium/hydrogen exchange
         Neurosciences
         Cell Biology
         Neurobiology
      image:
      isPartOf:
         name:Cellular and Molecular Neurobiology
         issn:
            1573-6830
            0272-4340
         volumeNumber:25
         type:
            Periodical
            PublicationVolume
      publisher:
         name:Kluwer Academic Publishers-Plenum Publishers
         logo:
            url:https://www.springernature.com/app-sn/public/images/logo-springernature.png
            type:ImageObject
         type:Organization
      author:
            name:IldikĂł Sipos
            affiliation:
                  name:Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences
                  address:
                     name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences, Hungary
                     type:PostalAddress
                  type:Organization
            type:Person
            name:BeĂĄta Tör’ócsik
            affiliation:
                  name:Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences
                  address:
                     name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences, Hungary
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Laszlo Tretter
            affiliation:
                  name:Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences
                  address:
                     name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences, Hungary
                     type:PostalAddress
                  type:Organization
            type:Person
            name:Vera Adam-Vizi
            affiliation:
                  name:Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences
                  address:
                     name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences, Hungary
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                  name:Semmelweis University
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         name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences, Hungary
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         name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences, Hungary
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      name:Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences
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         name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences, Hungary
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            address:
               name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences, Hungary
               type:PostalAddress
            type:Organization
      name:BeĂĄta Tör’ócsik
      affiliation:
            name:Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences
            address:
               name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences, Hungary
               type:PostalAddress
            type:Organization
      name:Laszlo Tretter
      affiliation:
            name:Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences
            address:
               name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences, Hungary
               type:PostalAddress
            type:Organization
      name:Vera Adam-Vizi
      affiliation:
            name:Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences
            address:
               name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences, Hungary
               type:PostalAddress
            type:Organization
            name:Semmelweis University
            address:
               name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary
               type:PostalAddress
            type:Organization
      email:[email protected]
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      name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences, Hungary
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      name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences, Hungary
      name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary and Neurochemistry Group, Hungarian Academy of Sciences, Hungary
      name:Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary
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