Mitochondrial complex i defect induces ROS release and degeneration in trabecular meshwork cells of POAG patients: Protection by antioxidants

Yuan He, Kar Wah Leung, Yue Hong Zhang, Shan Duan, Xiu Feng Zhong, Ru Zhang Jiang, Zhan Peng, Joyce Tombran-Tink, Jian Ge

Research output: Contribution to journalArticle

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Abstract

Purpose. There is growing evidence that oxidative stress contributes to the progression of primary open-angle glaucoma (POAG), a leading cause of irreversible blindness worldwide. The authors provide evidence that mitochondrial dysfunction is a possible mechanism for the loss of trabecular meshwork (TM) cells in persons with POAG. Methods. TM from patients with POAG (GTM) and age-matched subjects without disease (NTM) were obtained by standard surgical trabeculectomy. Primary TM cultures were treated with one of the following mitochondrial respiratory chain inhibitors: rotenone (ROT, complex I inhibitor), thenoyl-trifluoroacetone (TTFA, complex II inhibitor), myxothiazol or antimycin A (MYX, AM-complex III inhibitors); mitochondrial permeability transition (MPT) inhibitor cyclosporine A (CsA); and antioxidants vitamin E (Vit E) or N-acetylcysteine (NAC). Mitochondrial function was determined by changes in mito-chondrial membrane potential (δψm) and adenosine triphosphate (ATP) production with the fluorescent probes 5,5′6,6′ tetrachloro-1,1′ 3,3′-tetraethylbenzimid azolocarbocyanine iodide (JC-1) and a luciferin/luciferase-based ATP assay, respectively. Reactive oxygen species (ROS) level, determined by H 2-DCF-DA, and cell death' measured by lactate dehydrogenase activity and Annexin V-FITC labeling, were also examined. Results. GTM cells have higher endogenous ROS levels, lower ATP levels, and decreased Aψm and they are more sensitive to mitochondrial complex I inhibition than their normal counterparts. ROT induces a further increase in ROS production, the release of cytochrome c and decreases in ATP level and δψm in GTM cells, eventually leading to apoptosis. Complex II and III inhibition had little effect on the cells. Antioxidants protect against ROT-induced death by inhibiting ROS generation and cytochrome c release. Conclusions. The authors propose that a mitochondrial complex I defect is associated with the degeneration of TM cells in patients with POAG, and antioxidants and MPT inhibitors can reduce the progression of this condition.

Original languageEnglish (US)
Pages (from-to)1447-1458
Number of pages12
JournalInvestigative Ophthalmology and Visual Science
Volume49
Issue number4
DOIs
StatePublished - Apr 1 2008

Fingerprint

Trabecular Meshwork
Reactive Oxygen Species
Antioxidants
Adenosine Triphosphate
Cytochromes c
Permeability
Antimycin A
Rotenone
Trabeculectomy
Fluorescein-5-isothiocyanate
Electron Transport Complex III
Annexin A5
Acetylcysteine
Iodides
Blindness
Electron Transport
Luciferases
Vitamin E
Fluorescent Dyes
L-Lactate Dehydrogenase

All Science Journal Classification (ASJC) codes

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Cite this

He, Yuan ; Leung, Kar Wah ; Zhang, Yue Hong ; Duan, Shan ; Zhong, Xiu Feng ; Jiang, Ru Zhang ; Peng, Zhan ; Tombran-Tink, Joyce ; Ge, Jian. / Mitochondrial complex i defect induces ROS release and degeneration in trabecular meshwork cells of POAG patients : Protection by antioxidants. In: Investigative Ophthalmology and Visual Science. 2008 ; Vol. 49, No. 4. pp. 1447-1458.
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abstract = "Purpose. There is growing evidence that oxidative stress contributes to the progression of primary open-angle glaucoma (POAG), a leading cause of irreversible blindness worldwide. The authors provide evidence that mitochondrial dysfunction is a possible mechanism for the loss of trabecular meshwork (TM) cells in persons with POAG. Methods. TM from patients with POAG (GTM) and age-matched subjects without disease (NTM) were obtained by standard surgical trabeculectomy. Primary TM cultures were treated with one of the following mitochondrial respiratory chain inhibitors: rotenone (ROT, complex I inhibitor), thenoyl-trifluoroacetone (TTFA, complex II inhibitor), myxothiazol or antimycin A (MYX, AM-complex III inhibitors); mitochondrial permeability transition (MPT) inhibitor cyclosporine A (CsA); and antioxidants vitamin E (Vit E) or N-acetylcysteine (NAC). Mitochondrial function was determined by changes in mito-chondrial membrane potential (δψm) and adenosine triphosphate (ATP) production with the fluorescent probes 5,5′6,6′ tetrachloro-1,1′ 3,3′-tetraethylbenzimid azolocarbocyanine iodide (JC-1) and a luciferin/luciferase-based ATP assay, respectively. Reactive oxygen species (ROS) level, determined by H 2-DCF-DA, and cell death' measured by lactate dehydrogenase activity and Annexin V-FITC labeling, were also examined. Results. GTM cells have higher endogenous ROS levels, lower ATP levels, and decreased Aψm and they are more sensitive to mitochondrial complex I inhibition than their normal counterparts. ROT induces a further increase in ROS production, the release of cytochrome c and decreases in ATP level and δψm in GTM cells, eventually leading to apoptosis. Complex II and III inhibition had little effect on the cells. Antioxidants protect against ROT-induced death by inhibiting ROS generation and cytochrome c release. Conclusions. The authors propose that a mitochondrial complex I defect is associated with the degeneration of TM cells in patients with POAG, and antioxidants and MPT inhibitors can reduce the progression of this condition.",
author = "Yuan He and Leung, {Kar Wah} and Zhang, {Yue Hong} and Shan Duan and Zhong, {Xiu Feng} and Jiang, {Ru Zhang} and Zhan Peng and Joyce Tombran-Tink and Jian Ge",
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Mitochondrial complex i defect induces ROS release and degeneration in trabecular meshwork cells of POAG patients : Protection by antioxidants. / He, Yuan; Leung, Kar Wah; Zhang, Yue Hong; Duan, Shan; Zhong, Xiu Feng; Jiang, Ru Zhang; Peng, Zhan; Tombran-Tink, Joyce; Ge, Jian.

In: Investigative Ophthalmology and Visual Science, Vol. 49, No. 4, 01.04.2008, p. 1447-1458.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Mitochondrial complex i defect induces ROS release and degeneration in trabecular meshwork cells of POAG patients

T2 - Protection by antioxidants

AU - He, Yuan

AU - Leung, Kar Wah

AU - Zhang, Yue Hong

AU - Duan, Shan

AU - Zhong, Xiu Feng

AU - Jiang, Ru Zhang

AU - Peng, Zhan

AU - Tombran-Tink, Joyce

AU - Ge, Jian

PY - 2008/4/1

Y1 - 2008/4/1

N2 - Purpose. There is growing evidence that oxidative stress contributes to the progression of primary open-angle glaucoma (POAG), a leading cause of irreversible blindness worldwide. The authors provide evidence that mitochondrial dysfunction is a possible mechanism for the loss of trabecular meshwork (TM) cells in persons with POAG. Methods. TM from patients with POAG (GTM) and age-matched subjects without disease (NTM) were obtained by standard surgical trabeculectomy. Primary TM cultures were treated with one of the following mitochondrial respiratory chain inhibitors: rotenone (ROT, complex I inhibitor), thenoyl-trifluoroacetone (TTFA, complex II inhibitor), myxothiazol or antimycin A (MYX, AM-complex III inhibitors); mitochondrial permeability transition (MPT) inhibitor cyclosporine A (CsA); and antioxidants vitamin E (Vit E) or N-acetylcysteine (NAC). Mitochondrial function was determined by changes in mito-chondrial membrane potential (δψm) and adenosine triphosphate (ATP) production with the fluorescent probes 5,5′6,6′ tetrachloro-1,1′ 3,3′-tetraethylbenzimid azolocarbocyanine iodide (JC-1) and a luciferin/luciferase-based ATP assay, respectively. Reactive oxygen species (ROS) level, determined by H 2-DCF-DA, and cell death' measured by lactate dehydrogenase activity and Annexin V-FITC labeling, were also examined. Results. GTM cells have higher endogenous ROS levels, lower ATP levels, and decreased Aψm and they are more sensitive to mitochondrial complex I inhibition than their normal counterparts. ROT induces a further increase in ROS production, the release of cytochrome c and decreases in ATP level and δψm in GTM cells, eventually leading to apoptosis. Complex II and III inhibition had little effect on the cells. Antioxidants protect against ROT-induced death by inhibiting ROS generation and cytochrome c release. Conclusions. The authors propose that a mitochondrial complex I defect is associated with the degeneration of TM cells in patients with POAG, and antioxidants and MPT inhibitors can reduce the progression of this condition.

AB - Purpose. There is growing evidence that oxidative stress contributes to the progression of primary open-angle glaucoma (POAG), a leading cause of irreversible blindness worldwide. The authors provide evidence that mitochondrial dysfunction is a possible mechanism for the loss of trabecular meshwork (TM) cells in persons with POAG. Methods. TM from patients with POAG (GTM) and age-matched subjects without disease (NTM) were obtained by standard surgical trabeculectomy. Primary TM cultures were treated with one of the following mitochondrial respiratory chain inhibitors: rotenone (ROT, complex I inhibitor), thenoyl-trifluoroacetone (TTFA, complex II inhibitor), myxothiazol or antimycin A (MYX, AM-complex III inhibitors); mitochondrial permeability transition (MPT) inhibitor cyclosporine A (CsA); and antioxidants vitamin E (Vit E) or N-acetylcysteine (NAC). Mitochondrial function was determined by changes in mito-chondrial membrane potential (δψm) and adenosine triphosphate (ATP) production with the fluorescent probes 5,5′6,6′ tetrachloro-1,1′ 3,3′-tetraethylbenzimid azolocarbocyanine iodide (JC-1) and a luciferin/luciferase-based ATP assay, respectively. Reactive oxygen species (ROS) level, determined by H 2-DCF-DA, and cell death' measured by lactate dehydrogenase activity and Annexin V-FITC labeling, were also examined. Results. GTM cells have higher endogenous ROS levels, lower ATP levels, and decreased Aψm and they are more sensitive to mitochondrial complex I inhibition than their normal counterparts. ROT induces a further increase in ROS production, the release of cytochrome c and decreases in ATP level and δψm in GTM cells, eventually leading to apoptosis. Complex II and III inhibition had little effect on the cells. Antioxidants protect against ROT-induced death by inhibiting ROS generation and cytochrome c release. Conclusions. The authors propose that a mitochondrial complex I defect is associated with the degeneration of TM cells in patients with POAG, and antioxidants and MPT inhibitors can reduce the progression of this condition.

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