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Downregulation of Endogenous Hydrogen Sulfide Pathway Is Involved in Mitochondrion-Related Endothelial Cell Apoptosis Induced by High Salt.

Zong Y, Huang Y, Chen S, Zhu M, Chen Q, Feng S, Sun Y, Zhang Q, Tang C, Du J, Jin H - Oxid Med Cell Longev (2015)

Bottom Line: The cell apoptosis was detected by cell death ELISA and TdT-mediated dUTP nick end labeling (TUNEL) methods.However, supplementation of H2S donor markedly inhibited VEC oxidative stress and mitochondria-related VEC apoptosis induced by high salt.H2S/CSE pathway is an important endogenous defensive system in endothelial cells antagonizing high-salt insult.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Peking University First Hospital, Beijing 100034, China.

ABSTRACT

Background: The study aimed to investigate whether endogenous H2S pathway was involved in high-salt-stimulated mitochondria-related vascular endothelial cell (VEC) apoptosis.

Methods: Cultured human umbilical vein endothelial cells (HUVECs) were used in the study. H2S content in the supernatant was detected. Western blot was used to detect expression of cystathionine gamma-lyase (CSE), cleaved-caspase-3, and mitochondrial and cytosolic cytochrome c (cytc). Fluorescent probes were used to quantitatively detect superoxide anion generation and measure the in situ superoxide anion generation in HUVEC. Mitochondrial membrane pore opening, mitochondrial membrane potential, and caspase-9 activities were measured. The cell apoptosis was detected by cell death ELISA and TdT-mediated dUTP nick end labeling (TUNEL) methods.

Results: High-salt treatment downregulated the endogenous VEC H2S/CSE pathway, in association with increased generation of oxygen free radicals, decreased mitochondrial membrane potential, enhanced the opening of mitochondrial membrane permeability transition pore and leakage of mitochondrial cytc, activated cytoplasmic caspase-9 and caspase-3 and subsequently induced VEC apoptosis. However, supplementation of H2S donor markedly inhibited VEC oxidative stress and mitochondria-related VEC apoptosis induced by high salt.

Conclusion: H2S/CSE pathway is an important endogenous defensive system in endothelial cells antagonizing high-salt insult. The protective mechanisms for VEC damage might involve inhibiting oxidative stress and protecting mitochondrial injury.

No MeSH data available.


Related in: MedlinePlus

Changes in mitochondrial membrane potential and mitochondrial permeability transition pore (MPTP) opening in human umbilical vein endothelial cells (HUVECs). (a) Change of mitochondrial membrane potential detected by JC-1 fluorescent probe and examined by laser confocal microscope, with red fluorescence presenting JC-1 aggregate and green JC-1 monomer. (b) Changes of MPTP opening in HUVEC detected by calcein-AM as a fluorescence indicator by laser confocal microscopy. The green fluorescence quenching represented MPTP opening. Control: the cell treated with 137 mmol/L sodium. HS: high salt, the cell treated with 200 mmol/L sodium for 6 h. HS + NaHS: the cell pretreated with 200 μmol/L NaHS for 30 min following 200 mmol/L sodium for 6 h.
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fig3: Changes in mitochondrial membrane potential and mitochondrial permeability transition pore (MPTP) opening in human umbilical vein endothelial cells (HUVECs). (a) Change of mitochondrial membrane potential detected by JC-1 fluorescent probe and examined by laser confocal microscope, with red fluorescence presenting JC-1 aggregate and green JC-1 monomer. (b) Changes of MPTP opening in HUVEC detected by calcein-AM as a fluorescence indicator by laser confocal microscopy. The green fluorescence quenching represented MPTP opening. Control: the cell treated with 137 mmol/L sodium. HS: high salt, the cell treated with 200 mmol/L sodium for 6 h. HS + NaHS: the cell pretreated with 200 μmol/L NaHS for 30 min following 200 mmol/L sodium for 6 h.

Mentions: In HS group, the mitochondrial membrane potential was significantly reduced (Figure 3(a)) and mitochondrial permeability transition pore significantly opened (Figure 3(b)) relative to that of the control group, whereas H2S donor increased mitochondrial membrane potential (Figure 3(a)) and closed mitochondria permeability transition pore in HS-treated HUVEC (Figure 3(b)).


Downregulation of Endogenous Hydrogen Sulfide Pathway Is Involved in Mitochondrion-Related Endothelial Cell Apoptosis Induced by High Salt.

Zong Y, Huang Y, Chen S, Zhu M, Chen Q, Feng S, Sun Y, Zhang Q, Tang C, Du J, Jin H - Oxid Med Cell Longev (2015)

Changes in mitochondrial membrane potential and mitochondrial permeability transition pore (MPTP) opening in human umbilical vein endothelial cells (HUVECs). (a) Change of mitochondrial membrane potential detected by JC-1 fluorescent probe and examined by laser confocal microscope, with red fluorescence presenting JC-1 aggregate and green JC-1 monomer. (b) Changes of MPTP opening in HUVEC detected by calcein-AM as a fluorescence indicator by laser confocal microscopy. The green fluorescence quenching represented MPTP opening. Control: the cell treated with 137 mmol/L sodium. HS: high salt, the cell treated with 200 mmol/L sodium for 6 h. HS + NaHS: the cell pretreated with 200 μmol/L NaHS for 30 min following 200 mmol/L sodium for 6 h.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4442413&req=5

fig3: Changes in mitochondrial membrane potential and mitochondrial permeability transition pore (MPTP) opening in human umbilical vein endothelial cells (HUVECs). (a) Change of mitochondrial membrane potential detected by JC-1 fluorescent probe and examined by laser confocal microscope, with red fluorescence presenting JC-1 aggregate and green JC-1 monomer. (b) Changes of MPTP opening in HUVEC detected by calcein-AM as a fluorescence indicator by laser confocal microscopy. The green fluorescence quenching represented MPTP opening. Control: the cell treated with 137 mmol/L sodium. HS: high salt, the cell treated with 200 mmol/L sodium for 6 h. HS + NaHS: the cell pretreated with 200 μmol/L NaHS for 30 min following 200 mmol/L sodium for 6 h.
Mentions: In HS group, the mitochondrial membrane potential was significantly reduced (Figure 3(a)) and mitochondrial permeability transition pore significantly opened (Figure 3(b)) relative to that of the control group, whereas H2S donor increased mitochondrial membrane potential (Figure 3(a)) and closed mitochondria permeability transition pore in HS-treated HUVEC (Figure 3(b)).

Bottom Line: The cell apoptosis was detected by cell death ELISA and TdT-mediated dUTP nick end labeling (TUNEL) methods.However, supplementation of H2S donor markedly inhibited VEC oxidative stress and mitochondria-related VEC apoptosis induced by high salt.H2S/CSE pathway is an important endogenous defensive system in endothelial cells antagonizing high-salt insult.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Peking University First Hospital, Beijing 100034, China.

ABSTRACT

Background: The study aimed to investigate whether endogenous H2S pathway was involved in high-salt-stimulated mitochondria-related vascular endothelial cell (VEC) apoptosis.

Methods: Cultured human umbilical vein endothelial cells (HUVECs) were used in the study. H2S content in the supernatant was detected. Western blot was used to detect expression of cystathionine gamma-lyase (CSE), cleaved-caspase-3, and mitochondrial and cytosolic cytochrome c (cytc). Fluorescent probes were used to quantitatively detect superoxide anion generation and measure the in situ superoxide anion generation in HUVEC. Mitochondrial membrane pore opening, mitochondrial membrane potential, and caspase-9 activities were measured. The cell apoptosis was detected by cell death ELISA and TdT-mediated dUTP nick end labeling (TUNEL) methods.

Results: High-salt treatment downregulated the endogenous VEC H2S/CSE pathway, in association with increased generation of oxygen free radicals, decreased mitochondrial membrane potential, enhanced the opening of mitochondrial membrane permeability transition pore and leakage of mitochondrial cytc, activated cytoplasmic caspase-9 and caspase-3 and subsequently induced VEC apoptosis. However, supplementation of H2S donor markedly inhibited VEC oxidative stress and mitochondria-related VEC apoptosis induced by high salt.

Conclusion: H2S/CSE pathway is an important endogenous defensive system in endothelial cells antagonizing high-salt insult. The protective mechanisms for VEC damage might involve inhibiting oxidative stress and protecting mitochondrial injury.

No MeSH data available.


Related in: MedlinePlus