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Stanniocalcin-1 protects retinal ganglion cells by inhibiting apoptosis and oxidative damage.

Kim SJ, Ko JH, Yun JH, Kim JA, Kim TE, Lee HJ, Kim SH, Park KH, Oh JY - PLoS ONE (2013)

Bottom Line: We found that intravitreal injection of STC-1 increased the number of RGCs in the retina at days 7 and 14 after ONT, and decreased apoptosis and oxidative damage.In cultures, treatment with STC-1 dose-dependently increased cell viability, and decreased apoptosis and levels of reactive oxygen species in RGC-5 cells that were exposed to CoCl2.The expression of HIF-1α that was up-regulated by injury was significantly suppressed in the retina and in RGC-5 cells by STC-1 treatment.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Gangnam-gu, Seoul, Korea.

ABSTRACT
Optic neuropathy including glaucoma is one of the leading causes of irreversible vision loss, and there are currently no effective therapies. The hallmark of pathophysiology of optic neuropathy is oxidative stress and apoptotic death of retinal ganglion cells (RGCs), a population of neurons in the central nervous system with their soma in the inner retina and axons in the optic nerve. We here tested that an anti-apoptotic protein stanniocalcin-1 (STC-1) can prevent loss of RGCs in the rat retina with optic nerve transection (ONT) and in cultures of RGC-5 cells with CoCl2 injury. We found that intravitreal injection of STC-1 increased the number of RGCs in the retina at days 7 and 14 after ONT, and decreased apoptosis and oxidative damage. In cultures, treatment with STC-1 dose-dependently increased cell viability, and decreased apoptosis and levels of reactive oxygen species in RGC-5 cells that were exposed to CoCl2. The expression of HIF-1α that was up-regulated by injury was significantly suppressed in the retina and in RGC-5 cells by STC-1 treatment. The results suggested that intravitreal injection of STC-1 might be a useful therapy for optic nerve diseases in which RGCs undergo apoptosis through oxidative stress.

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STC-1 inhibited ROS levels in RGC-5 cells exposed to CoCl2.(A, B) Flow cytometry showed that CoCl2 significantly increased the percentage of cells positive for both CellROXTM and MitoTracker Green, a marker for oxidative stress, and treatment with either 100 or 500 ng/mL of STC-1 significantly decreased the percentages of CellROX+ MitoTracker Green+ cells in RGC-5 cells. (C) ELISA analysis for nitrotyrosine indicated that levels of nitrotyrosine were markedly increased in CoCl2-injured RGC-5 cells, and significantly decreased by STC-1 or N-acetylcysteine treatment. (D) Real time RT-PCR analysis indicated that expression of HIF-1α was induced in RGC-5 cells by CoCl2, and was significantly down-regulated by STC-1 (100 or 500 ng/mL). However, UCP2 transcripts were not increased by STC-1. (E) Western blot analysis for HIF-1α showed that HIF-1α protein was increased in RGC-5 cells after CoCl2 injury, and was decreased by STC-1 treatment. (F) ELISA showed that the levels of UCP2 protein were not increased in CoCl2-injured RGC-5 cells by STC-1 treatment, whereas N-acetylcysteine treatment significantly increased levels of UCP2. The values are presented as the mean ± SEM.
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pone-0063749-g004: STC-1 inhibited ROS levels in RGC-5 cells exposed to CoCl2.(A, B) Flow cytometry showed that CoCl2 significantly increased the percentage of cells positive for both CellROXTM and MitoTracker Green, a marker for oxidative stress, and treatment with either 100 or 500 ng/mL of STC-1 significantly decreased the percentages of CellROX+ MitoTracker Green+ cells in RGC-5 cells. (C) ELISA analysis for nitrotyrosine indicated that levels of nitrotyrosine were markedly increased in CoCl2-injured RGC-5 cells, and significantly decreased by STC-1 or N-acetylcysteine treatment. (D) Real time RT-PCR analysis indicated that expression of HIF-1α was induced in RGC-5 cells by CoCl2, and was significantly down-regulated by STC-1 (100 or 500 ng/mL). However, UCP2 transcripts were not increased by STC-1. (E) Western blot analysis for HIF-1α showed that HIF-1α protein was increased in RGC-5 cells after CoCl2 injury, and was decreased by STC-1 treatment. (F) ELISA showed that the levels of UCP2 protein were not increased in CoCl2-injured RGC-5 cells by STC-1 treatment, whereas N-acetylcysteine treatment significantly increased levels of UCP2. The values are presented as the mean ± SEM.

Mentions: We next evaluated the effect of STC-1 on ROS production in RGC-5 cells that were exposed to CoCl2. The percentage of cells that were both positive for CellROXTM and MitoTracker Green indicating production of mitochondrial ROS was increased by CoCl2, and reduced significantly by STC-1 treatment (Fig. 4A, B). Similarly, levels of nitrotyrosine, a marker of oxidative stress, were markedly increased in the cells by CoCl2 and significantly decreased by STC-1 or N-acetylcysteine (Fig. 4C). Together, data suggested that hypoxia induced by CoCl2 increased oxidative stress in RGC-5 cells, and STC-1 decreased oxidative stress. Also, similar to in vivo data (Fig 2D, E, F), the expression of HIF-1α was induced in RGC-5 cells by CoCl2 and significantly reduced by STC-1 both at transcript and protein levels (Fig. 4D, E). However, STC-1 treatment did not change the expression of UCP2 either at transcript or protein levels in RGC-5 cells, whereas N-acetylcysteine significantly increased UCP2 levels (Fig. 4D, F).


Stanniocalcin-1 protects retinal ganglion cells by inhibiting apoptosis and oxidative damage.

Kim SJ, Ko JH, Yun JH, Kim JA, Kim TE, Lee HJ, Kim SH, Park KH, Oh JY - PLoS ONE (2013)

STC-1 inhibited ROS levels in RGC-5 cells exposed to CoCl2.(A, B) Flow cytometry showed that CoCl2 significantly increased the percentage of cells positive for both CellROXTM and MitoTracker Green, a marker for oxidative stress, and treatment with either 100 or 500 ng/mL of STC-1 significantly decreased the percentages of CellROX+ MitoTracker Green+ cells in RGC-5 cells. (C) ELISA analysis for nitrotyrosine indicated that levels of nitrotyrosine were markedly increased in CoCl2-injured RGC-5 cells, and significantly decreased by STC-1 or N-acetylcysteine treatment. (D) Real time RT-PCR analysis indicated that expression of HIF-1α was induced in RGC-5 cells by CoCl2, and was significantly down-regulated by STC-1 (100 or 500 ng/mL). However, UCP2 transcripts were not increased by STC-1. (E) Western blot analysis for HIF-1α showed that HIF-1α protein was increased in RGC-5 cells after CoCl2 injury, and was decreased by STC-1 treatment. (F) ELISA showed that the levels of UCP2 protein were not increased in CoCl2-injured RGC-5 cells by STC-1 treatment, whereas N-acetylcysteine treatment significantly increased levels of UCP2. The values are presented as the mean ± SEM.
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Related In: Results  -  Collection

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pone-0063749-g004: STC-1 inhibited ROS levels in RGC-5 cells exposed to CoCl2.(A, B) Flow cytometry showed that CoCl2 significantly increased the percentage of cells positive for both CellROXTM and MitoTracker Green, a marker for oxidative stress, and treatment with either 100 or 500 ng/mL of STC-1 significantly decreased the percentages of CellROX+ MitoTracker Green+ cells in RGC-5 cells. (C) ELISA analysis for nitrotyrosine indicated that levels of nitrotyrosine were markedly increased in CoCl2-injured RGC-5 cells, and significantly decreased by STC-1 or N-acetylcysteine treatment. (D) Real time RT-PCR analysis indicated that expression of HIF-1α was induced in RGC-5 cells by CoCl2, and was significantly down-regulated by STC-1 (100 or 500 ng/mL). However, UCP2 transcripts were not increased by STC-1. (E) Western blot analysis for HIF-1α showed that HIF-1α protein was increased in RGC-5 cells after CoCl2 injury, and was decreased by STC-1 treatment. (F) ELISA showed that the levels of UCP2 protein were not increased in CoCl2-injured RGC-5 cells by STC-1 treatment, whereas N-acetylcysteine treatment significantly increased levels of UCP2. The values are presented as the mean ± SEM.
Mentions: We next evaluated the effect of STC-1 on ROS production in RGC-5 cells that were exposed to CoCl2. The percentage of cells that were both positive for CellROXTM and MitoTracker Green indicating production of mitochondrial ROS was increased by CoCl2, and reduced significantly by STC-1 treatment (Fig. 4A, B). Similarly, levels of nitrotyrosine, a marker of oxidative stress, were markedly increased in the cells by CoCl2 and significantly decreased by STC-1 or N-acetylcysteine (Fig. 4C). Together, data suggested that hypoxia induced by CoCl2 increased oxidative stress in RGC-5 cells, and STC-1 decreased oxidative stress. Also, similar to in vivo data (Fig 2D, E, F), the expression of HIF-1α was induced in RGC-5 cells by CoCl2 and significantly reduced by STC-1 both at transcript and protein levels (Fig. 4D, E). However, STC-1 treatment did not change the expression of UCP2 either at transcript or protein levels in RGC-5 cells, whereas N-acetylcysteine significantly increased UCP2 levels (Fig. 4D, F).

Bottom Line: We found that intravitreal injection of STC-1 increased the number of RGCs in the retina at days 7 and 14 after ONT, and decreased apoptosis and oxidative damage.In cultures, treatment with STC-1 dose-dependently increased cell viability, and decreased apoptosis and levels of reactive oxygen species in RGC-5 cells that were exposed to CoCl2.The expression of HIF-1α that was up-regulated by injury was significantly suppressed in the retina and in RGC-5 cells by STC-1 treatment.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Gangnam-gu, Seoul, Korea.

ABSTRACT
Optic neuropathy including glaucoma is one of the leading causes of irreversible vision loss, and there are currently no effective therapies. The hallmark of pathophysiology of optic neuropathy is oxidative stress and apoptotic death of retinal ganglion cells (RGCs), a population of neurons in the central nervous system with their soma in the inner retina and axons in the optic nerve. We here tested that an anti-apoptotic protein stanniocalcin-1 (STC-1) can prevent loss of RGCs in the rat retina with optic nerve transection (ONT) and in cultures of RGC-5 cells with CoCl2 injury. We found that intravitreal injection of STC-1 increased the number of RGCs in the retina at days 7 and 14 after ONT, and decreased apoptosis and oxidative damage. In cultures, treatment with STC-1 dose-dependently increased cell viability, and decreased apoptosis and levels of reactive oxygen species in RGC-5 cells that were exposed to CoCl2. The expression of HIF-1α that was up-regulated by injury was significantly suppressed in the retina and in RGC-5 cells by STC-1 treatment. The results suggested that intravitreal injection of STC-1 might be a useful therapy for optic nerve diseases in which RGCs undergo apoptosis through oxidative stress.

Show MeSH
Related in: MedlinePlus