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Bee Venom Protects against Rotenone-Induced Cell Death in NSC34 Motor Neuron Cells.

Jung SY, Lee KW, Choi SM, Yang EJ - Toxins (Basel) (2015)

Bottom Line: In this study, we tested the effects of BV on neuronal cell death by examining rotenone-induced mitochondrial dysfunction.Pretreatment with 2.5 μg/mL BV had a neuroprotective effect against 10 μM rotenone-induced cell death in NSC34 motor neuron cells.Pre-treatment with BV significantly enhanced cell viability and ameliorated mitochondrial impairment in rotenone-treated cellular model.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Research, Korea Institute of Oriental Medicine, 483 Expo-ro, Yuseong-gu, Daejeon 305-811, Korea. syzzim84@gmail.com.

ABSTRACT
Rotenone, an inhibitor of mitochondrial complex I of the mitochondrial respiratory chain, is known to elevate mitochondrial reactive oxygen species and induce apoptosis via activation of the caspase-3 pathway. Bee venom (BV) extracted from honey bees has been widely used in oriental medicine and contains melittin, apamin, adolapin, mast cell-degranulating peptide, and phospholipase A₂. In this study, we tested the effects of BV on neuronal cell death by examining rotenone-induced mitochondrial dysfunction. NSC34 motor neuron cells were pretreated with 2.5 μg/mL BV and stimulated with 10 μM rotenone to induce cell toxicity. We assessed cell death by Western blotting using specific antibodies, such as phospho-ERK1/2, phospho-JNK, and cleaved capase-3 and performed an MTT assay for evaluation of cell death and mitochondria staining. Pretreatment with 2.5 μg/mL BV had a neuroprotective effect against 10 μM rotenone-induced cell death in NSC34 motor neuron cells. Pre-treatment with BV significantly enhanced cell viability and ameliorated mitochondrial impairment in rotenone-treated cellular model. Moreover, BV treatment inhibited the activation of JNK signaling and cleaved caspase-3 related to cell death and increased ERK phosphorylation involved in cell survival in rotenone-treated NSC34 motor neuron cells. Taken together, we suggest that BV treatment can be useful for protection of neurons against oxidative stress or neurotoxin-induced cell death.

No MeSH data available.


Related in: MedlinePlus

Confocal microscopy images of mitochondria using Mitotracker® Red. Mitochondria stained with Mitotracker® Red in NSC 34 cells untreated (A), treated with 2.5 µg/mL BV for 24 h (B) or 10 µM rotenone for 24 h (C), pretreated with 2.5 µg/mL BV for 24 h and 10 µM rotenone for 24 h (D). Arrows indicate aggregated mitochondria. BV pretreatment reduces mictochondria impairment induced by rotenone treatment in NSC34 cells. The bar indicates 200 μm. (E) Quantification of the number of aggregated mitochondria in a cell of 10 microscopic visual fields randomly selected. The values shown are the means ±S.E.M. of data obtained from three independent experiments. *** and ### indicate p < 0.001. *** compared to Con and ### compared to Ro. Con: control, BV: BV-treated cell, Ro: rotenone-treated cell, and Ro + BV: BV pretreated-cells prior to rotenone treatment.
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toxins-07-03715-f004: Confocal microscopy images of mitochondria using Mitotracker® Red. Mitochondria stained with Mitotracker® Red in NSC 34 cells untreated (A), treated with 2.5 µg/mL BV for 24 h (B) or 10 µM rotenone for 24 h (C), pretreated with 2.5 µg/mL BV for 24 h and 10 µM rotenone for 24 h (D). Arrows indicate aggregated mitochondria. BV pretreatment reduces mictochondria impairment induced by rotenone treatment in NSC34 cells. The bar indicates 200 μm. (E) Quantification of the number of aggregated mitochondria in a cell of 10 microscopic visual fields randomly selected. The values shown are the means ±S.E.M. of data obtained from three independent experiments. *** and ### indicate p < 0.001. *** compared to Con and ### compared to Ro. Con: control, BV: BV-treated cell, Ro: rotenone-treated cell, and Ro + BV: BV pretreated-cells prior to rotenone treatment.

Mentions: To evaluate the effect of BV pretreatment on rotenone-induced mitochondria alteration, we stained neuronal cells with Mitotracker® Red probes that passively diffuse across the plasma membrane and accumulate mitochondria of live cells and observed mitochondria using confocal microscopy. As shown in Figure 4, mitochondria showed broad cytoplasmic distribution in both control and BV-treated NSC34 neuronal cells. However, 10 μM rotenone treatment for 24 h induced aggregated mitochondria in neuronal cells (Figure 4C). Interestingly, pretreatment with 2.5 μg/mL BV inhibited mitochondrial aggregation resulting from rotenone treatment (Figure 4D,E). These findings suggest that BV pretreatment can block interference with the electron transport chain in mitochondria induced by rotenone.


Bee Venom Protects against Rotenone-Induced Cell Death in NSC34 Motor Neuron Cells.

Jung SY, Lee KW, Choi SM, Yang EJ - Toxins (Basel) (2015)

Confocal microscopy images of mitochondria using Mitotracker® Red. Mitochondria stained with Mitotracker® Red in NSC 34 cells untreated (A), treated with 2.5 µg/mL BV for 24 h (B) or 10 µM rotenone for 24 h (C), pretreated with 2.5 µg/mL BV for 24 h and 10 µM rotenone for 24 h (D). Arrows indicate aggregated mitochondria. BV pretreatment reduces mictochondria impairment induced by rotenone treatment in NSC34 cells. The bar indicates 200 μm. (E) Quantification of the number of aggregated mitochondria in a cell of 10 microscopic visual fields randomly selected. The values shown are the means ±S.E.M. of data obtained from three independent experiments. *** and ### indicate p < 0.001. *** compared to Con and ### compared to Ro. Con: control, BV: BV-treated cell, Ro: rotenone-treated cell, and Ro + BV: BV pretreated-cells prior to rotenone treatment.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4591667&req=5

toxins-07-03715-f004: Confocal microscopy images of mitochondria using Mitotracker® Red. Mitochondria stained with Mitotracker® Red in NSC 34 cells untreated (A), treated with 2.5 µg/mL BV for 24 h (B) or 10 µM rotenone for 24 h (C), pretreated with 2.5 µg/mL BV for 24 h and 10 µM rotenone for 24 h (D). Arrows indicate aggregated mitochondria. BV pretreatment reduces mictochondria impairment induced by rotenone treatment in NSC34 cells. The bar indicates 200 μm. (E) Quantification of the number of aggregated mitochondria in a cell of 10 microscopic visual fields randomly selected. The values shown are the means ±S.E.M. of data obtained from three independent experiments. *** and ### indicate p < 0.001. *** compared to Con and ### compared to Ro. Con: control, BV: BV-treated cell, Ro: rotenone-treated cell, and Ro + BV: BV pretreated-cells prior to rotenone treatment.
Mentions: To evaluate the effect of BV pretreatment on rotenone-induced mitochondria alteration, we stained neuronal cells with Mitotracker® Red probes that passively diffuse across the plasma membrane and accumulate mitochondria of live cells and observed mitochondria using confocal microscopy. As shown in Figure 4, mitochondria showed broad cytoplasmic distribution in both control and BV-treated NSC34 neuronal cells. However, 10 μM rotenone treatment for 24 h induced aggregated mitochondria in neuronal cells (Figure 4C). Interestingly, pretreatment with 2.5 μg/mL BV inhibited mitochondrial aggregation resulting from rotenone treatment (Figure 4D,E). These findings suggest that BV pretreatment can block interference with the electron transport chain in mitochondria induced by rotenone.

Bottom Line: In this study, we tested the effects of BV on neuronal cell death by examining rotenone-induced mitochondrial dysfunction.Pretreatment with 2.5 μg/mL BV had a neuroprotective effect against 10 μM rotenone-induced cell death in NSC34 motor neuron cells.Pre-treatment with BV significantly enhanced cell viability and ameliorated mitochondrial impairment in rotenone-treated cellular model.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Research, Korea Institute of Oriental Medicine, 483 Expo-ro, Yuseong-gu, Daejeon 305-811, Korea. syzzim84@gmail.com.

ABSTRACT
Rotenone, an inhibitor of mitochondrial complex I of the mitochondrial respiratory chain, is known to elevate mitochondrial reactive oxygen species and induce apoptosis via activation of the caspase-3 pathway. Bee venom (BV) extracted from honey bees has been widely used in oriental medicine and contains melittin, apamin, adolapin, mast cell-degranulating peptide, and phospholipase A₂. In this study, we tested the effects of BV on neuronal cell death by examining rotenone-induced mitochondrial dysfunction. NSC34 motor neuron cells were pretreated with 2.5 μg/mL BV and stimulated with 10 μM rotenone to induce cell toxicity. We assessed cell death by Western blotting using specific antibodies, such as phospho-ERK1/2, phospho-JNK, and cleaved capase-3 and performed an MTT assay for evaluation of cell death and mitochondria staining. Pretreatment with 2.5 μg/mL BV had a neuroprotective effect against 10 μM rotenone-induced cell death in NSC34 motor neuron cells. Pre-treatment with BV significantly enhanced cell viability and ameliorated mitochondrial impairment in rotenone-treated cellular model. Moreover, BV treatment inhibited the activation of JNK signaling and cleaved caspase-3 related to cell death and increased ERK phosphorylation involved in cell survival in rotenone-treated NSC34 motor neuron cells. Taken together, we suggest that BV treatment can be useful for protection of neurons against oxidative stress or neurotoxin-induced cell death.

No MeSH data available.


Related in: MedlinePlus