Mitochondria-Targeted Peptide MTP-131 Alleviates Mitochondrial Dysfunction and Oxidative Damage in Human Trabecular Meshwork Cells

Abstract

PURPOSE.To investigate the antioxidative ability of a novel mitochondria-targeted peptide MTP-131 in immortalized human trabecular meshwork (iHTM) and glaucomatous human trabecular meshwork (GTM 3 ) cell lines. METHODS. Cultured iHTM and GTM 3 cells were pretreated with MTP-131 for 1 hour, and sustained oxidative stress was induced by subjecting TM cells to 200 M hydrogen peroxide (H 2 O 2 ) for 24 hours. Untreated cells and cells incubated with H 2 O 2 alone were used as controls. Lactate dehydrogenase (LDH) assay was used to determine cell viability. Changes of mitochondrial membrane potential (⌬⌿m) and generation of intracellular reactive oxygen species (ROS) were analyzed by flow cytometry and confocal microscopy. Activation of caspase 3 was quantified by Western blotting, and apoptosis was measured by flow cytometry. Release of cytochrome c and changes in cytoskeleton were analyzed by confocal microscopy. Data were analyzed with commercial data analysis software and P Ͻ 0.05 was considered to be statistically significant. RESULTS. In both iHTM and GTM 3 cells, decrease of ⌬⌿m and elevation of intracellular ROS were detected after sustained oxidative stress induced by H 2 O 2 . When cells were pretreated with MTP-131, the H 2 O 2 -induced mitochondrial depolarization was prevented; intracellular ROS, LDH release, and apoptosis were significantly decreased; release of cytochrome c from mitochondria to cytoplasm and activation of caspase 3 were inhibited. In addition, cytoskeleton changes caused by H 2 O 2 were also alleviated by MTP-131. CONCLUSIONS. Mitochondria-targeted peptide MTP-131 could prevent both iHTM and GTM 3 cells from sustained oxidative stress induced by H 2 O 2 . (Invest Ophthalmol Vis Sci. 2011;52: 7027-7037)

Mitochondria-targeted peptides prevent on contrast-induced acute kidney injury in the rats with hypercholesterolemia

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Endoplasmic reticulum stress promotes amyloid-beta peptides production in RGC-5 cells

“…Intracellular ROS was detected by using oxidation-sensitive fluorescent probe 2′,7′-dichlorodihydrofluorescein diacetate (H 2 DCFDA), as described previously (Chen et al 2011). Cells were harvested at 12, 24, and 48 h after tunicamycin or vehicle treatment, incubated with 1 μM H 2 DCFDA at 37ºC for 30 min in the dark, rinsed twice with PBS, and analyzed immediately by Becton Dickinson FACSAria™ flow cytometer (BD Biosciences, San Jose, CA, USA) using excitation and emission wavelengths of 488 and 530 nm, respectively.…”

Mitochondria-Targeted Antioxidant Peptide SS31 Protects Cultured Human Lens Epithelial Cells against Oxidative Stress

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