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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

BV inhibits the expression of cleaved caspase-3 in NSC34 cells. (A) Expression of cleaved caspase-3 proteins was detected in rotenone-induced NSC34 cells. NSC34 cells incubated with 10 µM rotenone for the indicated time periods. The loading control for the cell lysates was determined by re-probing the membranes with α-tubulin antibody; (B) BV attenuates the expression level of cleaved caspase-3 protein in NSC34 neuronal cells. NSC34 cells were pretreated with 2.5 µg/mL BV for 24 h and then stimulated with 10 μM rotenone for 24 h in the presence or absence of BV; (C) BV inhibits the expression of cleaved caspase-3 protein in rotenone-treated in primary cortical neuronal cells system. BV pretreated with 2.5 µg/mL for 24 h and stimulated with 10 μM rotenone for 24 h in the presence or absence of BV in primary cortical neuronal cells. The loading control for the cell lysates was determined by re-probing the membranes with α-tubulin antibody. The values shown are the means ±S.E.M. of data obtained from three independent experiments. *p < 0.05, **p < 0.01, *** compared to * and ** and indicates p < 0.001.
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toxins-07-03715-f003: BV inhibits the expression of cleaved caspase-3 in NSC34 cells. (A) Expression of cleaved caspase-3 proteins was detected in rotenone-induced NSC34 cells. NSC34 cells incubated with 10 µM rotenone for the indicated time periods. The loading control for the cell lysates was determined by re-probing the membranes with α-tubulin antibody; (B) BV attenuates the expression level of cleaved caspase-3 protein in NSC34 neuronal cells. NSC34 cells were pretreated with 2.5 µg/mL BV for 24 h and then stimulated with 10 μM rotenone for 24 h in the presence or absence of BV; (C) BV inhibits the expression of cleaved caspase-3 protein in rotenone-treated in primary cortical neuronal cells system. BV pretreated with 2.5 µg/mL for 24 h and stimulated with 10 μM rotenone for 24 h in the presence or absence of BV in primary cortical neuronal cells. The loading control for the cell lysates was determined by re-probing the membranes with α-tubulin antibody. The values shown are the means ±S.E.M. of data obtained from three independent experiments. *p < 0.05, **p < 0.01, *** compared to * and ** and indicates p < 0.001.

Mentions: To investigate the effects of BV on oxidative stress-induced mitochondria dysfunction, capase-3 activation was assessed after exposure to rotenone in NSC34 neuronal cells. The expression level of cleaved caspase-3 was significantly increased after 6 h of 10 μM rotenone treatment compared to untreated NSC34 neuronal cells (Figure 3A). However, pretreatment with 2.5 μg/mL BV suppressed by two-fold the increase of caspase-3 activation induced by rotenone treatment compared with rotenone-treated NSC34 cells (Figure 3B,C). To confirm BV effects against rotenone, we performed the same experiment with primary cortical neuronal cells. Consistent with the result in NSC34 cell line, 10 μM rotenone treatment induced the activation of caspase-3 but 2.5 μg/mL BV treatment reduced rotenone-induced cleaved caspase-3 expression. Those results suggest that BV pretreatment could prevent oxidative stress-induced mitochondria dysfunction.


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

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

BV inhibits the expression of cleaved caspase-3 in NSC34 cells. (A) Expression of cleaved caspase-3 proteins was detected in rotenone-induced NSC34 cells. NSC34 cells incubated with 10 µM rotenone for the indicated time periods. The loading control for the cell lysates was determined by re-probing the membranes with α-tubulin antibody; (B) BV attenuates the expression level of cleaved caspase-3 protein in NSC34 neuronal cells. NSC34 cells were pretreated with 2.5 µg/mL BV for 24 h and then stimulated with 10 μM rotenone for 24 h in the presence or absence of BV; (C) BV inhibits the expression of cleaved caspase-3 protein in rotenone-treated in primary cortical neuronal cells system. BV pretreated with 2.5 µg/mL for 24 h and stimulated with 10 μM rotenone for 24 h in the presence or absence of BV in primary cortical neuronal cells. The loading control for the cell lysates was determined by re-probing the membranes with α-tubulin antibody. The values shown are the means ±S.E.M. of data obtained from three independent experiments. *p < 0.05, **p < 0.01, *** compared to * and ** and indicates p < 0.001.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591667&req=5

toxins-07-03715-f003: BV inhibits the expression of cleaved caspase-3 in NSC34 cells. (A) Expression of cleaved caspase-3 proteins was detected in rotenone-induced NSC34 cells. NSC34 cells incubated with 10 µM rotenone for the indicated time periods. The loading control for the cell lysates was determined by re-probing the membranes with α-tubulin antibody; (B) BV attenuates the expression level of cleaved caspase-3 protein in NSC34 neuronal cells. NSC34 cells were pretreated with 2.5 µg/mL BV for 24 h and then stimulated with 10 μM rotenone for 24 h in the presence or absence of BV; (C) BV inhibits the expression of cleaved caspase-3 protein in rotenone-treated in primary cortical neuronal cells system. BV pretreated with 2.5 µg/mL for 24 h and stimulated with 10 μM rotenone for 24 h in the presence or absence of BV in primary cortical neuronal cells. The loading control for the cell lysates was determined by re-probing the membranes with α-tubulin antibody. The values shown are the means ±S.E.M. of data obtained from three independent experiments. *p < 0.05, **p < 0.01, *** compared to * and ** and indicates p < 0.001.
Mentions: To investigate the effects of BV on oxidative stress-induced mitochondria dysfunction, capase-3 activation was assessed after exposure to rotenone in NSC34 neuronal cells. The expression level of cleaved caspase-3 was significantly increased after 6 h of 10 μM rotenone treatment compared to untreated NSC34 neuronal cells (Figure 3A). However, pretreatment with 2.5 μg/mL BV suppressed by two-fold the increase of caspase-3 activation induced by rotenone treatment compared with rotenone-treated NSC34 cells (Figure 3B,C). To confirm BV effects against rotenone, we performed the same experiment with primary cortical neuronal cells. Consistent with the result in NSC34 cell line, 10 μM rotenone treatment induced the activation of caspase-3 but 2.5 μg/mL BV treatment reduced rotenone-induced cleaved caspase-3 expression. Those results suggest that BV pretreatment could prevent oxidative stress-induced mitochondria dysfunction.

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