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Neuroprotective Effects of Alpha-Mangostin on MPP(+)-Induced Apoptotic Cell Death in Neuroblastoma SH-SY5Y Cells.

Janhom P, Dharmasaroja P - J Toxicol (2015)

Bottom Line: Alpha-mangostin reduced ROS formation induced by MPP(+).The cotreated cells showed a significant decrease in activated caspase-3 compared with MPP(+) treatment alone.Our data suggest that cytoprotection of alpha-mangostin against MPP(+)-induced apoptosis may be associated with the reduction of ROS production, modulating the balance of pro- and antiapoptotic genes, and suppression of caspase-3 activation.

View Article: PubMed Central - PubMed

Affiliation: Toxicology Graduate Program, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.

ABSTRACT
In vitro studies have shown that extracts from mangosteen (Garcinia mangostana Linn.) act as antioxidants and cytoprotective agents against oxidative damage. The protective effect of alpha-mangostin, the major xanthone found in the pericarp of the mangosteen, in cellular models of Parkinson's disease (PD), has not been investigated. This study aims to investigate whether alpha-mangostin could protect SH-SY5Y neuroblastoma cells from MPP(+)-induced apoptosis. The effects of alpha-mangostin on MPP(+)-induced cell death were evaluated with a cell viability assay, staining for nuclear DNA morphology, flow cytometry for apoptotic cells and reactive oxygen species (ROS) production, quantitative real-time PCR for the expression of p53, Bax, and Bcl-2, and western blot analysis for cleaved caspase-3. Concomitant treatment with alpha-mangostin attenuated the effect of MPP(+) on cell viability and apoptotic cell death. Alpha-mangostin reduced ROS formation induced by MPP(+). Bax/Bcl-2 expression ratio and expression of p53 were significantly lower in cells cocultured with alpha-mangostin and MPP(+). The cotreated cells showed a significant decrease in activated caspase-3 compared with MPP(+) treatment alone. Our data suggest that cytoprotection of alpha-mangostin against MPP(+)-induced apoptosis may be associated with the reduction of ROS production, modulating the balance of pro- and antiapoptotic genes, and suppression of caspase-3 activation.

No MeSH data available.


Related in: MedlinePlus

Effect of alpha-mangostin on Bax, Bcl-2, and p53 mRNA expression in MPP+-treated SH-SY5Y cells. Cells were treated with 10 μM alpha-mangostin (α-M), 1000 μM MPP+, or the combination of α-M and MPP+ for 24 hours. Expression of Bax (a), Bcl-2 (b), Bax/Bcl-2 ratio (c), and p53 mRNA (d) was analyzed with quantitative real-time RT-PCR, relatively compared to their respective untreated controls. The expression levels of the target gene were estimated by the 2−ΔΔCt method after normalizing to the expression level of β-actin. Data are expressed as mean ± SEM (n = 3).  ∗P < 0.01;  ∗∗P < 0.001.
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fig7: Effect of alpha-mangostin on Bax, Bcl-2, and p53 mRNA expression in MPP+-treated SH-SY5Y cells. Cells were treated with 10 μM alpha-mangostin (α-M), 1000 μM MPP+, or the combination of α-M and MPP+ for 24 hours. Expression of Bax (a), Bcl-2 (b), Bax/Bcl-2 ratio (c), and p53 mRNA (d) was analyzed with quantitative real-time RT-PCR, relatively compared to their respective untreated controls. The expression levels of the target gene were estimated by the 2−ΔΔCt method after normalizing to the expression level of β-actin. Data are expressed as mean ± SEM (n = 3).  ∗P < 0.01;  ∗∗P < 0.001.

Mentions: Expression of mRNA of Bax and Bcl-2 was investigated using real-time quantitative RT-PCR analysis. Bax expression significantly increased in 1000 μm MPP+-treated cells, compared to that of control cells (Figure 7(a)). Cotreatment of 10 μm alpha-mangostin and 1000 μm MPP+ for 24 hours significantly decreased the Bax expression, compared to MPP+ treatment alone (P < 0.01). Treatment with 1000 μm MPP+ significantly decreased Bcl-2 expression, while cotreatment with 10 μm alpha-mangostin significantly increased Bcl-2 expression (P < 0.01; Figure 7(b)). As a result, Bax/Bcl-2 expression was significantly increased in SH-SY5Y cells treated with 1000 μm MPP+ for 24 hours (P < 0.001; Figure 7(c)), whereas Bax/Bcl-2 expression was significantly lowered in cells cocultured in 10 μm alpha-mangostin and 1000 μm MPP+ (P < 0.001). An upstream regulation of Bax and Bcl-2 was then investigated. The expression of p53 mRNA was significantly increased in 1000 μm MPP+-treated SH-SY5Y cells after 24-hour exposure when compared to unexposed cells (P < 0.001; Figure 7(d)). Cotreatment of 10 μm alpha-mangostin and 1000 μm MPP+ significantly reduced p53 expression when compared to cells cultured in MPP+ alone (P < 0.01).


Neuroprotective Effects of Alpha-Mangostin on MPP(+)-Induced Apoptotic Cell Death in Neuroblastoma SH-SY5Y Cells.

Janhom P, Dharmasaroja P - J Toxicol (2015)

Effect of alpha-mangostin on Bax, Bcl-2, and p53 mRNA expression in MPP+-treated SH-SY5Y cells. Cells were treated with 10 μM alpha-mangostin (α-M), 1000 μM MPP+, or the combination of α-M and MPP+ for 24 hours. Expression of Bax (a), Bcl-2 (b), Bax/Bcl-2 ratio (c), and p53 mRNA (d) was analyzed with quantitative real-time RT-PCR, relatively compared to their respective untreated controls. The expression levels of the target gene were estimated by the 2−ΔΔCt method after normalizing to the expression level of β-actin. Data are expressed as mean ± SEM (n = 3).  ∗P < 0.01;  ∗∗P < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4556078&req=5

fig7: Effect of alpha-mangostin on Bax, Bcl-2, and p53 mRNA expression in MPP+-treated SH-SY5Y cells. Cells were treated with 10 μM alpha-mangostin (α-M), 1000 μM MPP+, or the combination of α-M and MPP+ for 24 hours. Expression of Bax (a), Bcl-2 (b), Bax/Bcl-2 ratio (c), and p53 mRNA (d) was analyzed with quantitative real-time RT-PCR, relatively compared to their respective untreated controls. The expression levels of the target gene were estimated by the 2−ΔΔCt method after normalizing to the expression level of β-actin. Data are expressed as mean ± SEM (n = 3).  ∗P < 0.01;  ∗∗P < 0.001.
Mentions: Expression of mRNA of Bax and Bcl-2 was investigated using real-time quantitative RT-PCR analysis. Bax expression significantly increased in 1000 μm MPP+-treated cells, compared to that of control cells (Figure 7(a)). Cotreatment of 10 μm alpha-mangostin and 1000 μm MPP+ for 24 hours significantly decreased the Bax expression, compared to MPP+ treatment alone (P < 0.01). Treatment with 1000 μm MPP+ significantly decreased Bcl-2 expression, while cotreatment with 10 μm alpha-mangostin significantly increased Bcl-2 expression (P < 0.01; Figure 7(b)). As a result, Bax/Bcl-2 expression was significantly increased in SH-SY5Y cells treated with 1000 μm MPP+ for 24 hours (P < 0.001; Figure 7(c)), whereas Bax/Bcl-2 expression was significantly lowered in cells cocultured in 10 μm alpha-mangostin and 1000 μm MPP+ (P < 0.001). An upstream regulation of Bax and Bcl-2 was then investigated. The expression of p53 mRNA was significantly increased in 1000 μm MPP+-treated SH-SY5Y cells after 24-hour exposure when compared to unexposed cells (P < 0.001; Figure 7(d)). Cotreatment of 10 μm alpha-mangostin and 1000 μm MPP+ significantly reduced p53 expression when compared to cells cultured in MPP+ alone (P < 0.01).

Bottom Line: Alpha-mangostin reduced ROS formation induced by MPP(+).The cotreated cells showed a significant decrease in activated caspase-3 compared with MPP(+) treatment alone.Our data suggest that cytoprotection of alpha-mangostin against MPP(+)-induced apoptosis may be associated with the reduction of ROS production, modulating the balance of pro- and antiapoptotic genes, and suppression of caspase-3 activation.

View Article: PubMed Central - PubMed

Affiliation: Toxicology Graduate Program, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.

ABSTRACT
In vitro studies have shown that extracts from mangosteen (Garcinia mangostana Linn.) act as antioxidants and cytoprotective agents against oxidative damage. The protective effect of alpha-mangostin, the major xanthone found in the pericarp of the mangosteen, in cellular models of Parkinson's disease (PD), has not been investigated. This study aims to investigate whether alpha-mangostin could protect SH-SY5Y neuroblastoma cells from MPP(+)-induced apoptosis. The effects of alpha-mangostin on MPP(+)-induced cell death were evaluated with a cell viability assay, staining for nuclear DNA morphology, flow cytometry for apoptotic cells and reactive oxygen species (ROS) production, quantitative real-time PCR for the expression of p53, Bax, and Bcl-2, and western blot analysis for cleaved caspase-3. Concomitant treatment with alpha-mangostin attenuated the effect of MPP(+) on cell viability and apoptotic cell death. Alpha-mangostin reduced ROS formation induced by MPP(+). Bax/Bcl-2 expression ratio and expression of p53 were significantly lower in cells cocultured with alpha-mangostin and MPP(+). The cotreated cells showed a significant decrease in activated caspase-3 compared with MPP(+) treatment alone. Our data suggest that cytoprotection of alpha-mangostin against MPP(+)-induced apoptosis may be associated with the reduction of ROS production, modulating the balance of pro- and antiapoptotic genes, and suppression of caspase-3 activation.

No MeSH data available.


Related in: MedlinePlus