Yi Li1, Yi Jia, Jun Zhou, Kaixun Huang. 1. Hubei Key Laboratory of Bioinorganic Chemistry & Materia Medica, School of Chemistry and Chemical Engineering, Huazhong University of Science and Technology, Hubei 430074, PR China; Department of Biochemistry and Molecular Biology, Chongqing Medical University, Chongqing, 400016, PR China.
Abstract
AIMS: To determine roles of methionine sulfoxide reductase B1 (MsrB1) in protecting lens mitochondria against oxidative damage, the influences of MsrB1 gene silencing on high-glucose-induced apoptosis in human lens epithelial (HLE) cells were studied. MAIN METHODS: Our study used four groups of cells: normal control, MsrB1 gene silenced, high glucose (30mM) exposed and MsrB1 gene silenced cells followed with high glucose exposure. In all cases we detected cell viability, cell apoptosis rate, intracellular reactive oxygen species (ROS) and malondialdehyde (MDA) levels, alteration of mitochondrial membrane potential, release of mitochondrial cytochrome c as well as an increase in activity of caspase-3. KEY FINDINGS: The results showed that MsrB1 gene silencing by short interfering RNA (siRNA) in HLE cells clearly resulted in oxidative stress, decrease in mitochondrial membrane potential and release of mitochondrial cytochrome c as well as an increase in activity of caspase-3 and the percentage of apoptotic cells. When MsrB1-silenced HLE cells were exposed to high glucose, characteristic of high-glucose-induced mitochondrial dysfunctions were further exacerbated. SIGNIFICANCE: MsrB1 plays important roles in protecting HLE cell mitochondria against oxidative damage and inhibits oxidative stress-induced apoptosis in diabetic cataracts by scavenging ROS.
AIMS: To determine roles of methionine sulfoxide reductase B1 (MsrB1) in protecting lens mitochondria against oxidative damage, the influences of MsrB1 gene silencing on high-glucose-induced apoptosis in human lens epithelial (HLE) cells were studied. MAIN METHODS: Our study used four groups of cells: normal control, MsrB1 gene silenced, high glucose (30mM) exposed and MsrB1 gene silenced cells followed with high glucose exposure. In all cases we detected cell viability, cell apoptosis rate, intracellular reactive oxygen species (ROS) and malondialdehyde (MDA) levels, alteration of mitochondrial membrane potential, release of mitochondrial cytochrome c as well as an increase in activity of caspase-3. KEY FINDINGS: The results showed that MsrB1 gene silencing by short interfering RNA (siRNA) in HLE cells clearly resulted in oxidative stress, decrease in mitochondrial membrane potential and release of mitochondrial cytochrome c as well as an increase in activity of caspase-3 and the percentage of apoptotic cells. When MsrB1-silenced HLE cells were exposed to high glucose, characteristic of high-glucose-induced mitochondrial dysfunctions were further exacerbated. SIGNIFICANCE: MsrB1 plays important roles in protecting HLE cell mitochondria against oxidative damage and inhibits oxidative stress-induced apoptosis in diabetic cataracts by scavenging ROS.