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Artemisinin protects PC12 cells against β -amyloid-induced apoptosis through activation of the ERK1/2 signaling pathway

View Article: PubMed Central - PubMed

ABSTRACT

Accumulating evidence displays that an abnormal deposition of amyloid beta-peptide (Aβ) is the primary cause of the pathogenesis of Alzheimer's disease (AD). And therefore the elimination of Aβ is regarded as an important strategy for AD treatment. The discovery of drug candidates using culture neuronal cells against Aβ peptide toxicity is believed to be an effective approach to develop drug for the treatment of AD patients. We have previously showed that artemisinin, a FDA-approved anti-malaria drug, has neuroprotective effects recently. In the present study, we aimed to investigate the effects and potential mechanism of artemisinin in protecting neuronal PC12 cells from toxicity of β amyloid peptide. Our studies revealed that artemisinin, in clinical relevant concentration, protected and rescued PC12 cells from Aβ25–35-induced cell death. Further study showed that artemisinin significantly ameliorated cell death due to Aβ25–35 insult by restoring abnormal changes in nuclear morphology, lactate dehydrogenase, intracellular ROS, mitochondrial membrane potential and activity of apoptotic caspase. Western blotting analysis demonstrated that artemisinin activated extracellular regulated kinase ERK1/2 but not Akt survival signaling. Consistent with the role of ERK1/2, preincubation of cells with ERK1/2 pathway inhibitor PD98059 blocked the effect of artemisinin while PI3K inhibitor LY294002 has no effect. Moreover, Aβ1-42 also caused cells death of PC12 cells while artemisinin suppressed Aβ1-42 cytotoxicity in PC12 cells. Taken together, these results, at the first time, suggest that artemisinin is a potential protectant against β amyloid insult through activation of the ERK1/2 pathway. Our finding provides a potential application of artemisinin in prevention and treatment of AD.

No MeSH data available.


Artemisinin concentration-dependently suppressed Aβ25-35-induced cell viability lose in PC12 cells. (A) The structure of artemisinin. (B) Cells were treated with Aβ25-35 (0.03–10 μM) or 0.1% DMSO (vehicle control) for 24 h and cell viability was measured using the MTT assay(N=3). (C) Cells were pre-treated with artemisinin (3,125–100 μM) or 0.1% DMSO (vehicle control) for 1 h and then incubated with or without 0.3 μM Aβ25-35 for a further 24 h and cell viability were measured by MTT assay (N=3). (D) PC12 cells were incubated with 0.1, 0.3, 1 μM Aβ25-35 for 30 min and post-treated with artemisinin (25 or 50 μM) for 24 h and cell viability were measured by MTT assay (N=3). *P<0.05, **P<0.01,***P<0.005 versus the control or Aβ25-35 treated group as indicated; ###P<0.005 versus control group was considered statistically significant differences.
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f0005: Artemisinin concentration-dependently suppressed Aβ25-35-induced cell viability lose in PC12 cells. (A) The structure of artemisinin. (B) Cells were treated with Aβ25-35 (0.03–10 μM) or 0.1% DMSO (vehicle control) for 24 h and cell viability was measured using the MTT assay(N=3). (C) Cells were pre-treated with artemisinin (3,125–100 μM) or 0.1% DMSO (vehicle control) for 1 h and then incubated with or without 0.3 μM Aβ25-35 for a further 24 h and cell viability were measured by MTT assay (N=3). (D) PC12 cells were incubated with 0.1, 0.3, 1 μM Aβ25-35 for 30 min and post-treated with artemisinin (25 or 50 μM) for 24 h and cell viability were measured by MTT assay (N=3). *P<0.05, **P<0.01,***P<0.005 versus the control or Aβ25-35 treated group as indicated; ###P<0.005 versus control group was considered statistically significant differences.

Mentions: Aβ-induced apoptosis in PC12 cells was a common and reliable cellular toxicity model for AD related studies in vitro [23], [24], [25]. We first tested the cytotoxicity of the most usual used peptide Aβ25–35 (active component of Aβ peptides) on PC12 cells by MTT assay in our laboratory conditions. As shown in Fig. 1B, exposure of cells to different concentrations of Aβ25–35 for 24 h resulted in a notable decrease of the cell viability in a concentration-dependent manner, which indicated that Aβ25–35 could induce toxicity in PC12 cells. The toxicity was observed in the lowest dose of 0.03 μM and maximal in 3–10 μM. 0.3 μM Aβ25–35 was chosen in following experiment because of the 30–40% cell death was observed.


Artemisinin protects PC12 cells against β -amyloid-induced apoptosis through activation of the ERK1/2 signaling pathway
Artemisinin concentration-dependently suppressed Aβ25-35-induced cell viability lose in PC12 cells. (A) The structure of artemisinin. (B) Cells were treated with Aβ25-35 (0.03–10 μM) or 0.1% DMSO (vehicle control) for 24 h and cell viability was measured using the MTT assay(N=3). (C) Cells were pre-treated with artemisinin (3,125–100 μM) or 0.1% DMSO (vehicle control) for 1 h and then incubated with or without 0.3 μM Aβ25-35 for a further 24 h and cell viability were measured by MTT assay (N=3). (D) PC12 cells were incubated with 0.1, 0.3, 1 μM Aβ25-35 for 30 min and post-treated with artemisinin (25 or 50 μM) for 24 h and cell viability were measured by MTT assay (N=3). *P<0.05, **P<0.01,***P<0.005 versus the control or Aβ25-35 treated group as indicated; ###P<0.005 versus control group was considered statistically significant differences.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

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Show All Figures
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f0005: Artemisinin concentration-dependently suppressed Aβ25-35-induced cell viability lose in PC12 cells. (A) The structure of artemisinin. (B) Cells were treated with Aβ25-35 (0.03–10 μM) or 0.1% DMSO (vehicle control) for 24 h and cell viability was measured using the MTT assay(N=3). (C) Cells were pre-treated with artemisinin (3,125–100 μM) or 0.1% DMSO (vehicle control) for 1 h and then incubated with or without 0.3 μM Aβ25-35 for a further 24 h and cell viability were measured by MTT assay (N=3). (D) PC12 cells were incubated with 0.1, 0.3, 1 μM Aβ25-35 for 30 min and post-treated with artemisinin (25 or 50 μM) for 24 h and cell viability were measured by MTT assay (N=3). *P<0.05, **P<0.01,***P<0.005 versus the control or Aβ25-35 treated group as indicated; ###P<0.005 versus control group was considered statistically significant differences.
Mentions: Aβ-induced apoptosis in PC12 cells was a common and reliable cellular toxicity model for AD related studies in vitro [23], [24], [25]. We first tested the cytotoxicity of the most usual used peptide Aβ25–35 (active component of Aβ peptides) on PC12 cells by MTT assay in our laboratory conditions. As shown in Fig. 1B, exposure of cells to different concentrations of Aβ25–35 for 24 h resulted in a notable decrease of the cell viability in a concentration-dependent manner, which indicated that Aβ25–35 could induce toxicity in PC12 cells. The toxicity was observed in the lowest dose of 0.03 μM and maximal in 3–10 μM. 0.3 μM Aβ25–35 was chosen in following experiment because of the 30–40% cell death was observed.

View Article: PubMed Central - PubMed

ABSTRACT

Accumulating evidence displays that an abnormal deposition of amyloid beta-peptide (A&beta;) is the primary cause of the pathogenesis of Alzheimer's disease (AD). And therefore the elimination of A&beta; is regarded as an important strategy for AD treatment. The discovery of drug candidates using culture neuronal cells against A&beta; peptide toxicity is believed to be an effective approach to develop drug for the treatment of AD patients. We have previously showed that artemisinin, a FDA-approved anti-malaria drug, has neuroprotective effects recently. In the present study, we aimed to investigate the effects and potential mechanism of artemisinin in protecting neuronal PC12 cells from toxicity of &beta; amyloid peptide. Our studies revealed that artemisinin, in clinical relevant concentration, protected and rescued PC12 cells from A&beta;25&ndash;35-induced cell death. Further study showed that artemisinin significantly ameliorated cell death due to A&beta;25&ndash;35 insult by restoring abnormal changes in nuclear morphology, lactate dehydrogenase, intracellular ROS, mitochondrial membrane potential and activity of apoptotic caspase. Western blotting analysis demonstrated that artemisinin activated extracellular regulated kinase ERK1/2 but not Akt survival signaling. Consistent with the role of ERK1/2, preincubation of cells with ERK1/2 pathway inhibitor PD98059 blocked the effect of artemisinin while PI3K inhibitor LY294002 has no effect. Moreover, A&beta;1-42 also caused cells death of PC12 cells while artemisinin suppressed A&beta;1-42 cytotoxicity in PC12 cells. Taken together, these results, at the first time, suggest that artemisinin is a potential protectant against &beta; amyloid insult through activation of the ERK1/2 pathway. Our finding provides a potential application of artemisinin in prevention and treatment of AD.

No MeSH data available.