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Carnosic Acid, a Rosemary Phenolic Compound, Induces Apoptosis Through Reactive Oxygen Species-Mediated p38 Activation in Human Neuroblastoma IMR-32 Cells

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Abstract

Carnosic acid (CA), a rosemary phenolic compound, has been shown to display anti-cancer activity. We examined the apoptotic effect of CA in human neuroblastoma IMR-32 cells and elucidated the role of the reactive oxygen species (ROS) and mitogen-activated protein kinase (MAPK) associated with carcinogenesis. The result indicated that CA decreased the cell viability in a dose-dependent manner. Further investigation in IMR-32 cells revealed that cell apoptosis following CA treatment is the mechanism as confirmed by flow cytometry, hoechst 33258, and caspase-3/-9 and poly(ADP-ribose) polymerase (PARP) activation. Immunoblotting suggested a down-regulation of anti-apoptotic Bcl-2 protein in the CA-treated cells. In flow cytometric analysis, CA caused the generation of reactive oxygen species (ROS); however, pretreatment with the antioxidant N-acetylcysteine (NAC) attenuated the CA-induced generation of ROS and apoptosis. This effect was accompanied by increased activation of p38 and by decreased activation of extracellular signal-regulated kinase (ERK) as well as activation of c-Jun NH2-terminal kinase (JNK). Moreover, NAC attenuated the CA-induced phosphorylation of p38. Silencing of p38 by siRNA gene knockdown reduced the CA-induced activation of caspase-3. In conclusion, ROS-mediated p38 MAPK activation plays a critical role in CA-induced apoptosis in IMR-32 cells.

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Abbreviations

CA:

Carnosic acid

JNK:

c-Jun NH2-terminal kinase

ERK:

Extracellular signal-regulated kinase

MAPK:

Mitogen-activated protein kinase

MTT:

3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolim bromide

NAC:

N-acetylcysteine

PARP:

poly(ADP-ribose) polymerase

PI:

Propidium iodide

ROS:

Reactive oxygen species

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Acknowledgment

This work was supported by China Medical University (CMU) (grant no. CMU97-246).

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Correspondence to Chia-Wen Tsai.

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Tsai, CW., Lin, CY., Lin, HH. et al. Carnosic Acid, a Rosemary Phenolic Compound, Induces Apoptosis Through Reactive Oxygen Species-Mediated p38 Activation in Human Neuroblastoma IMR-32 Cells. Neurochem Res 36, 2442–2451 (2011). https://doi.org/10.1007/s11064-011-0573-4

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