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Protective effects of SS31 on t‑BHP induced oxidative damage in 661W cells.

Ma W, Zhu X, Ding X, Li T, Hu Y, Hu X, Yuan L, Lei L, Hu A, Luo Y, Tang S - Mol Med Rep (2015)

Bottom Line: The viability of the cells improved following treatment with SS31 between 100 nM and 1 µM, compared with untreated control group.Compared with the t‑BHP treatment group (20.0±3.8%), the number of annexin V‑positive cells decreased dose‑dependently to 13.6±2.6, 9.8±0.5 and 7.4±2.0% in the SS‑31 treated group at concentrations of 10 nM, 100 nM and 1 µM, respectively.Treatment with SS‑31 significantly prevented the t‑BHP‑induced expression of nitrotyrosine and 8‑OHdG, decreased the quantity of mitochondrial ROS, increased mitochondrial potential, and prevented the release of cytochrome c from mitochondria into the cytoplasm.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat‑Sen University, Guangzhou, Guangdong 510060, P.R. China.

ABSTRACT
The present study aimed to investigate the ability of SS31, a novel mitochondria‑targeted peptide to protect against t‑BHP‑induced mitochondrial dysfunction and apoptosis in 661W cell lines. The 661W cells were treated with various concentrations of SS‑31 and an MTT assay was used to determine cell viability. The expression of nitrotyrosine and 8‑hydroxydeoxyguanosine (8‑OHdG) was detected using immunofluorescent staining. Apoptosis were assessed using Hoechst staining and an annexin V/propidium iodide flow cytometer. Reactive oxygen species (ROS) were detected using MitoSOXTM with confocal microscopy. Changes in mitochondrial membrane potential were analyzed using flow cytometry. In addition, the release of cytochrome c was analyzed using confocal microscopy. The viability of the cells improved following treatment with SS31 between 100 nM and 1 µM, compared with untreated control group. Compared with the t‑BHP treatment group (20.0±3.8%), the number of annexin V‑positive cells decreased dose‑dependently to 13.6±2.6, 9.8±0.5 and 7.4±2.0% in the SS‑31 treated group at concentrations of 10 nM, 100 nM and 1 µM, respectively. Treatment with SS‑31 significantly prevented the t‑BHP‑induced expression of nitrotyrosine and 8‑OHdG, decreased the quantity of mitochondrial ROS, increased mitochondrial potential, and prevented the release of cytochrome c from mitochondria into the cytoplasm. Therefore, the SS31 mitochondria‑targeted peptide protected the 661W cells from the sustained oxidative stress induced by t‑BHP.

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SS31 prevents the decrease in 661W cell viability induced by oxida-tive damage. (A) Concentration-dependent decrease in 661W cell viability with increasing t-BHP after 24 h incubation (*P<0.001, vs. control). In all cases, the control indicates untreated 661W cells. (B) Inhibition of the t-BHP -induced decrease in cell viability by SS31 in the 661W cells. Cell viability improved following treatment with SS31 (10 nM-1 µM) for 24 h (*P<0.01, vs. t-BHP). Data are presented as the mean ± standard error of the mean.
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f1-mmr-12-04-5026: SS31 prevents the decrease in 661W cell viability induced by oxida-tive damage. (A) Concentration-dependent decrease in 661W cell viability with increasing t-BHP after 24 h incubation (*P<0.001, vs. control). In all cases, the control indicates untreated 661W cells. (B) Inhibition of the t-BHP -induced decrease in cell viability by SS31 in the 661W cells. Cell viability improved following treatment with SS31 (10 nM-1 µM) for 24 h (*P<0.01, vs. t-BHP). Data are presented as the mean ± standard error of the mean.

Mentions: The viability of the 661W cells was reduced following exposure to t-BHP for 24 h in a dose-dependent manner. Marked cytotoxicity was observed at concentrations of ≥100 µM t-BHP, compared with the untreated control cells (Fig. 1A). As shown in Fig. 1B, treatment of the 661W cells with t-BHP at 100 µM for 24 h led to ~30% loss of cell viability (P<0.01; n=4). By contrast, treatment with SS31 led to increased cell survival (P<0.01; n-4), however, concentrations <10 nM did not improve 661W cell survival following oxidant injury.


Protective effects of SS31 on t‑BHP induced oxidative damage in 661W cells.

Ma W, Zhu X, Ding X, Li T, Hu Y, Hu X, Yuan L, Lei L, Hu A, Luo Y, Tang S - Mol Med Rep (2015)

SS31 prevents the decrease in 661W cell viability induced by oxida-tive damage. (A) Concentration-dependent decrease in 661W cell viability with increasing t-BHP after 24 h incubation (*P<0.001, vs. control). In all cases, the control indicates untreated 661W cells. (B) Inhibition of the t-BHP -induced decrease in cell viability by SS31 in the 661W cells. Cell viability improved following treatment with SS31 (10 nM-1 µM) for 24 h (*P<0.01, vs. t-BHP). Data are presented as the mean ± standard error of the mean.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581771&req=5

f1-mmr-12-04-5026: SS31 prevents the decrease in 661W cell viability induced by oxida-tive damage. (A) Concentration-dependent decrease in 661W cell viability with increasing t-BHP after 24 h incubation (*P<0.001, vs. control). In all cases, the control indicates untreated 661W cells. (B) Inhibition of the t-BHP -induced decrease in cell viability by SS31 in the 661W cells. Cell viability improved following treatment with SS31 (10 nM-1 µM) for 24 h (*P<0.01, vs. t-BHP). Data are presented as the mean ± standard error of the mean.
Mentions: The viability of the 661W cells was reduced following exposure to t-BHP for 24 h in a dose-dependent manner. Marked cytotoxicity was observed at concentrations of ≥100 µM t-BHP, compared with the untreated control cells (Fig. 1A). As shown in Fig. 1B, treatment of the 661W cells with t-BHP at 100 µM for 24 h led to ~30% loss of cell viability (P<0.01; n=4). By contrast, treatment with SS31 led to increased cell survival (P<0.01; n-4), however, concentrations <10 nM did not improve 661W cell survival following oxidant injury.

Bottom Line: The viability of the cells improved following treatment with SS31 between 100 nM and 1 µM, compared with untreated control group.Compared with the t‑BHP treatment group (20.0±3.8%), the number of annexin V‑positive cells decreased dose‑dependently to 13.6±2.6, 9.8±0.5 and 7.4±2.0% in the SS‑31 treated group at concentrations of 10 nM, 100 nM and 1 µM, respectively.Treatment with SS‑31 significantly prevented the t‑BHP‑induced expression of nitrotyrosine and 8‑OHdG, decreased the quantity of mitochondrial ROS, increased mitochondrial potential, and prevented the release of cytochrome c from mitochondria into the cytoplasm.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat‑Sen University, Guangzhou, Guangdong 510060, P.R. China.

ABSTRACT
The present study aimed to investigate the ability of SS31, a novel mitochondria‑targeted peptide to protect against t‑BHP‑induced mitochondrial dysfunction and apoptosis in 661W cell lines. The 661W cells were treated with various concentrations of SS‑31 and an MTT assay was used to determine cell viability. The expression of nitrotyrosine and 8‑hydroxydeoxyguanosine (8‑OHdG) was detected using immunofluorescent staining. Apoptosis were assessed using Hoechst staining and an annexin V/propidium iodide flow cytometer. Reactive oxygen species (ROS) were detected using MitoSOXTM with confocal microscopy. Changes in mitochondrial membrane potential were analyzed using flow cytometry. In addition, the release of cytochrome c was analyzed using confocal microscopy. The viability of the cells improved following treatment with SS31 between 100 nM and 1 µM, compared with untreated control group. Compared with the t‑BHP treatment group (20.0±3.8%), the number of annexin V‑positive cells decreased dose‑dependently to 13.6±2.6, 9.8±0.5 and 7.4±2.0% in the SS‑31 treated group at concentrations of 10 nM, 100 nM and 1 µM, respectively. Treatment with SS‑31 significantly prevented the t‑BHP‑induced expression of nitrotyrosine and 8‑OHdG, decreased the quantity of mitochondrial ROS, increased mitochondrial potential, and prevented the release of cytochrome c from mitochondria into the cytoplasm. Therefore, the SS31 mitochondria‑targeted peptide protected the 661W cells from the sustained oxidative stress induced by t‑BHP.

Show MeSH
Related in: MedlinePlus