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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 suppressed Aβ25-35-induced LDH release and apoptosis in PC12 cells. After pre-treatment with 25 μM artemisinin or 0.1% DMSO (vehicle control) for 1 h, PC12 cells were incubated with or without 0.3 μM Aβ25-35 for another 24 h the release of LDH (A) was measured by LDH assay and the apoptosis cells (N=3) (B) was detected by staining with Hoechst 33342 and visualized by fluorescence microscopy. The number of apoptotic nuclei with condensed chromatin (C) was counted from the photomicrographs and presented as a percentage of the total number of nuclei (N=3). ###P<0.005 versus control group; *P<0.05, **P<0.005 versus the Aβ-treated group were considered statistically significant differences.
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f0010: Artemisinin suppressed Aβ25-35-induced LDH release and apoptosis in PC12 cells. After pre-treatment with 25 μM artemisinin or 0.1% DMSO (vehicle control) for 1 h, PC12 cells were incubated with or without 0.3 μM Aβ25-35 for another 24 h the release of LDH (A) was measured by LDH assay and the apoptosis cells (N=3) (B) was detected by staining with Hoechst 33342 and visualized by fluorescence microscopy. The number of apoptotic nuclei with condensed chromatin (C) was counted from the photomicrographs and presented as a percentage of the total number of nuclei (N=3). ###P<0.005 versus control group; *P<0.05, **P<0.005 versus the Aβ-treated group were considered statistically significant differences.

Mentions: To evaluate the protective effects of artemisinin, PC12 cells were treated with artemisinin for 1 h before exposure to Aβ25-35 for 24 h. The result of MTT assay (Fig. 1C) revealed that the treatment of 0.3 μM Aβ25-35 resulted in dominant cell death (43%), whereas pretreatment with 12.5 and 25 μM of artemisinin significantly attenuated Aβ25-35- induced cell death. To clarify whether artemisinin could rescue cell from toxicity of Aβ25-35, 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. The results (Fig. 1D) demonstrated that artemisinin not only can protected but also rescue PC12 cells against Aβ25-35-induced cell death. The protective activity of artemisinin was also confirmed by the lactate dehydrogenase (LDH) assay. As shown in Fig. 2A, pre-treated cells with 25 μM of artemisinin for 1 h significantly reduced Aβ25-35-induced LDH leakage (from 160% to 135%). Nuclei condensation was observed in PC12 cells after exposure to Aβ25-35 in Hoechst 33342 staining assay (Fig. 2B). However, a pretreatment of 25 μM artemisinin definitely improved these changes (from 36% to 23%) (Fig. 2C).


Artemisinin protects PC12 cells against β -amyloid-induced apoptosis through activation of the ERK1/2 signaling pathway
Artemisinin suppressed Aβ25-35-induced LDH release and apoptosis in PC12 cells. After pre-treatment with 25 μM artemisinin or 0.1% DMSO (vehicle control) for 1 h, PC12 cells were incubated with or without 0.3 μM Aβ25-35 for another 24 h the release of LDH (A) was measured by LDH assay and the apoptosis cells (N=3) (B) was detected by staining with Hoechst 33342 and visualized by fluorescence microscopy. The number of apoptotic nuclei with condensed chromatin (C) was counted from the photomicrographs and presented as a percentage of the total number of nuclei (N=3). ###P<0.005 versus control group; *P<0.05, **P<0.005 versus the Aβ-treated group were considered statistically significant differences.
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Related In: Results  -  Collection

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Show All Figures
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f0010: Artemisinin suppressed Aβ25-35-induced LDH release and apoptosis in PC12 cells. After pre-treatment with 25 μM artemisinin or 0.1% DMSO (vehicle control) for 1 h, PC12 cells were incubated with or without 0.3 μM Aβ25-35 for another 24 h the release of LDH (A) was measured by LDH assay and the apoptosis cells (N=3) (B) was detected by staining with Hoechst 33342 and visualized by fluorescence microscopy. The number of apoptotic nuclei with condensed chromatin (C) was counted from the photomicrographs and presented as a percentage of the total number of nuclei (N=3). ###P<0.005 versus control group; *P<0.05, **P<0.005 versus the Aβ-treated group were considered statistically significant differences.
Mentions: To evaluate the protective effects of artemisinin, PC12 cells were treated with artemisinin for 1 h before exposure to Aβ25-35 for 24 h. The result of MTT assay (Fig. 1C) revealed that the treatment of 0.3 μM Aβ25-35 resulted in dominant cell death (43%), whereas pretreatment with 12.5 and 25 μM of artemisinin significantly attenuated Aβ25-35- induced cell death. To clarify whether artemisinin could rescue cell from toxicity of Aβ25-35, 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. The results (Fig. 1D) demonstrated that artemisinin not only can protected but also rescue PC12 cells against Aβ25-35-induced cell death. The protective activity of artemisinin was also confirmed by the lactate dehydrogenase (LDH) assay. As shown in Fig. 2A, pre-treated cells with 25 μM of artemisinin for 1 h significantly reduced Aβ25-35-induced LDH leakage (from 160% to 135%). Nuclei condensation was observed in PC12 cells after exposure to Aβ25-35 in Hoechst 33342 staining assay (Fig. 2B). However, a pretreatment of 25 μM artemisinin definitely improved these changes (from 36% to 23%) (Fig. 2C).

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.