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Protective Effect of Total Phenolic Compounds from Inula helenium on Hydrogen Peroxide-induced Oxidative Stress in SH-SY5Y Cells.

Wang J, Zhao YM, Zhang B, Guo CY - Indian J Pharm Sci (2015 Mar-Apr)

Bottom Line: It was shown that hydrogen peroxide significantly induced the loss of cell viability, increment of apoptosis, formation of reactive oxygen species, reduction of superoxide dismutase activity, decrease in mitochondrial membrane potential and a decrease in adenosine triphosphate production.On the other hand, total phenolic compounds dose-dependently reversed these effects.This study suggests that total phenolic compounds exert neuroprotective effects against hydrogen peroxide-induced oxidative damage via blocking reactive oxygen species production and improving mitochondrial function.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, HeBei North University, Zhangjiakou, 075000, China.

ABSTRACT
Inula helenium has been reported to contain a large amount of phenolic compounds, which have shown promise in scavenging free radicals and prevention of neurodegenerative diseases. This study is to investigate the neuroprotective effects of total phenolic compounds from I. helenium on hydrogen peroxide-induced oxidative damage in human SH-SY5Y cells. Antioxidant capacity of total phenolic compounds was determined by radical scavenging activity, the level of intracellular reactive oxygen species and superoxide dismutase activity. The cytotoxicity of total phenolic compounds was determined using a cell counting kit-8 assay. The effect of total phenolic compounds on cell apoptosis due to hydrogen peroxide-induced oxidative damage was detected by Hoechst 33258 and Annexin-V/PI staining using fluorescence microscope and flow cytometry, respectively. Mitochondrial function was evaluated using the mitochondrial membrane potential and mitochondrial ATP synthesis by JC-1 dye and high performance liquid chromatography, respectively. It was shown that hydrogen peroxide significantly induced the loss of cell viability, increment of apoptosis, formation of reactive oxygen species, reduction of superoxide dismutase activity, decrease in mitochondrial membrane potential and a decrease in adenosine triphosphate production. On the other hand, total phenolic compounds dose-dependently reversed these effects. This study suggests that total phenolic compounds exert neuroprotective effects against hydrogen peroxide-induced oxidative damage via blocking reactive oxygen species production and improving mitochondrial function. The potential of total phenolic compounds and its neuroprotective mechanisms in attenuating hydrogen peroxide-induced oxidative stress-related cytotoxicity is worth further exploration.

No MeSH data available.


Related in: MedlinePlus

TPC suppresses the apoptosis of SH-SY5Y cells induced by H2O2 by flow cytometric analysis.The SH-SY5Y cells pre-treated with total phenolic compounds (TPC) for 1 h were exposed to 200 μM H2O2 for 24 h and then the cells were labeled with Annexin-V/PI staining and analyzed by flow cytometer. (a) Flow cytometric histograms of control, H2O2, and cells pretreated with 0.5 and 5 μg/ml TPC. (b) Fluorescence intensities of control group, H2O2 and H2O2 in cells pretreated with 0.5 and 5 μg/ml TPC. Each bar represents mean±SD of 3 estimations, **P<0.05 vs control group and ##P<0.05 vs H2O2 alone group.
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Figure 4: TPC suppresses the apoptosis of SH-SY5Y cells induced by H2O2 by flow cytometric analysis.The SH-SY5Y cells pre-treated with total phenolic compounds (TPC) for 1 h were exposed to 200 μM H2O2 for 24 h and then the cells were labeled with Annexin-V/PI staining and analyzed by flow cytometer. (a) Flow cytometric histograms of control, H2O2, and cells pretreated with 0.5 and 5 μg/ml TPC. (b) Fluorescence intensities of control group, H2O2 and H2O2 in cells pretreated with 0.5 and 5 μg/ml TPC. Each bar represents mean±SD of 3 estimations, **P<0.05 vs control group and ##P<0.05 vs H2O2 alone group.

Mentions: Apoptotic and necrotic changes in the cells were determined morphologically by flow cytometry with Annexin V-FITC/PI staining. The effects of TPC on early and late apoptosis/necrosis induced by H2O2 were detected using Annexin V/PI staining as previously described[27]. FITC-conjugated Annexin V is a marker for apoptosis and PI is an indicator of necrotic cells[28]. Treated cells with H2O2 significantly increased the percentage of apoptotic and necrotic cells. Simultaneous TPC pretreatment of cells significantly reduced the number of cells labeled with FITC-conjugated Annexin V and decreased markedly the percentage of apoptotic cells (fig. 4).


Protective Effect of Total Phenolic Compounds from Inula helenium on Hydrogen Peroxide-induced Oxidative Stress in SH-SY5Y Cells.

Wang J, Zhao YM, Zhang B, Guo CY - Indian J Pharm Sci (2015 Mar-Apr)

TPC suppresses the apoptosis of SH-SY5Y cells induced by H2O2 by flow cytometric analysis.The SH-SY5Y cells pre-treated with total phenolic compounds (TPC) for 1 h were exposed to 200 μM H2O2 for 24 h and then the cells were labeled with Annexin-V/PI staining and analyzed by flow cytometer. (a) Flow cytometric histograms of control, H2O2, and cells pretreated with 0.5 and 5 μg/ml TPC. (b) Fluorescence intensities of control group, H2O2 and H2O2 in cells pretreated with 0.5 and 5 μg/ml TPC. Each bar represents mean±SD of 3 estimations, **P<0.05 vs control group and ##P<0.05 vs H2O2 alone group.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4442464&req=5

Figure 4: TPC suppresses the apoptosis of SH-SY5Y cells induced by H2O2 by flow cytometric analysis.The SH-SY5Y cells pre-treated with total phenolic compounds (TPC) for 1 h were exposed to 200 μM H2O2 for 24 h and then the cells were labeled with Annexin-V/PI staining and analyzed by flow cytometer. (a) Flow cytometric histograms of control, H2O2, and cells pretreated with 0.5 and 5 μg/ml TPC. (b) Fluorescence intensities of control group, H2O2 and H2O2 in cells pretreated with 0.5 and 5 μg/ml TPC. Each bar represents mean±SD of 3 estimations, **P<0.05 vs control group and ##P<0.05 vs H2O2 alone group.
Mentions: Apoptotic and necrotic changes in the cells were determined morphologically by flow cytometry with Annexin V-FITC/PI staining. The effects of TPC on early and late apoptosis/necrosis induced by H2O2 were detected using Annexin V/PI staining as previously described[27]. FITC-conjugated Annexin V is a marker for apoptosis and PI is an indicator of necrotic cells[28]. Treated cells with H2O2 significantly increased the percentage of apoptotic and necrotic cells. Simultaneous TPC pretreatment of cells significantly reduced the number of cells labeled with FITC-conjugated Annexin V and decreased markedly the percentage of apoptotic cells (fig. 4).

Bottom Line: It was shown that hydrogen peroxide significantly induced the loss of cell viability, increment of apoptosis, formation of reactive oxygen species, reduction of superoxide dismutase activity, decrease in mitochondrial membrane potential and a decrease in adenosine triphosphate production.On the other hand, total phenolic compounds dose-dependently reversed these effects.This study suggests that total phenolic compounds exert neuroprotective effects against hydrogen peroxide-induced oxidative damage via blocking reactive oxygen species production and improving mitochondrial function.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacy, HeBei North University, Zhangjiakou, 075000, China.

ABSTRACT
Inula helenium has been reported to contain a large amount of phenolic compounds, which have shown promise in scavenging free radicals and prevention of neurodegenerative diseases. This study is to investigate the neuroprotective effects of total phenolic compounds from I. helenium on hydrogen peroxide-induced oxidative damage in human SH-SY5Y cells. Antioxidant capacity of total phenolic compounds was determined by radical scavenging activity, the level of intracellular reactive oxygen species and superoxide dismutase activity. The cytotoxicity of total phenolic compounds was determined using a cell counting kit-8 assay. The effect of total phenolic compounds on cell apoptosis due to hydrogen peroxide-induced oxidative damage was detected by Hoechst 33258 and Annexin-V/PI staining using fluorescence microscope and flow cytometry, respectively. Mitochondrial function was evaluated using the mitochondrial membrane potential and mitochondrial ATP synthesis by JC-1 dye and high performance liquid chromatography, respectively. It was shown that hydrogen peroxide significantly induced the loss of cell viability, increment of apoptosis, formation of reactive oxygen species, reduction of superoxide dismutase activity, decrease in mitochondrial membrane potential and a decrease in adenosine triphosphate production. On the other hand, total phenolic compounds dose-dependently reversed these effects. This study suggests that total phenolic compounds exert neuroprotective effects against hydrogen peroxide-induced oxidative damage via blocking reactive oxygen species production and improving mitochondrial function. The potential of total phenolic compounds and its neuroprotective mechanisms in attenuating hydrogen peroxide-induced oxidative stress-related cytotoxicity is worth further exploration.

No MeSH data available.


Related in: MedlinePlus