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Neuritogenic effect of standardized extract of Centella asiatica ECa233 on human neuroblastoma cells.

Wanakhachornkrai O, Pongrakhananon V, Chunhacha P, Wanasuntronwong A, Vattanajun A, Tantisira B, Chanvorachote P, Tantisira MH - BMC Complement Altern Med (2013)

Bottom Line: While ECa 233 had only limited effects on cell viability, it significantly enhanced neurite outgrowth of IMR-32 cells at the concentrations of 1-100 μg/ml.ERK1/2 and Akt signaling pathways seemed to account for the neurotrophic effect observed.In conjunction with in vivo neuroprotective effect of ECa 233 previously reported, the results obtained support further development of ECa 233 for clinical use in neuronal injury or neurodegenerative diseases.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Background: In order to gain insight into neuroprotective effects of ECa 233, a standardized extract of Centella asiatica, previously demonstrated in animal models of memory impairment induced by transient global ischemia or intracerebroventricular injection of β-amyloid, the effect of ECa 233 on neurite outgrowth of human IMR-32 neuroblastoma cell line was investigated.

Methods: Cells were seeded and incubated with various concentrations of ECa 233. Morphometric analysis was carried out by a measurement of the longest neurite growth of cells at 24 and 48 h. Contributing signaling pathways possibly involved were subsequently elucidated by western blot analysis.

Results: While ECa 233 had only limited effects on cell viability, it significantly enhanced neurite outgrowth of IMR-32 cells at the concentrations of 1-100 μg/ml. Western blot analysis revealed that ECa 233 significantly upregulated the level of activated ERK1/2 and Akt of the treated cells suggesting their involvement in the neuritogenic effect observed, which was subsequently verified by the finding that an addition of their respective inhibitors could reverse the effect of ECa 233 on these cells.

Conclusions: The present study clearly demonstrated neurite outgrowth promoting activity of ECa 233. ERK1/2 and Akt signaling pathways seemed to account for the neurotrophic effect observed. In conjunction with in vivo neuroprotective effect of ECa 233 previously reported, the results obtained support further development of ECa 233 for clinical use in neuronal injury or neurodegenerative diseases.

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Morphometric analysis of IMR-32 cells affected by ECa 233. After cells cultured for 24 and 48 h in the presence of either ECa 233 (0.1, 1, 10 and 100 μg/ml) or BDNF (100 ng/ml). Morphometric analysis was carried out by measuring neurites from 60 living cells per treated condition, (n = 3). Average of the neurite length was significantly increased after being treated with ECa 233 (1, 10 and 100 μg/ml) or BDNF. The data presented as mean ± S.E. ** = p < 0.01, *** = p < 0.001 vs non-treated control.
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Figure 3: Morphometric analysis of IMR-32 cells affected by ECa 233. After cells cultured for 24 and 48 h in the presence of either ECa 233 (0.1, 1, 10 and 100 μg/ml) or BDNF (100 ng/ml). Morphometric analysis was carried out by measuring neurites from 60 living cells per treated condition, (n = 3). Average of the neurite length was significantly increased after being treated with ECa 233 (1, 10 and 100 μg/ml) or BDNF. The data presented as mean ± S.E. ** = p < 0.01, *** = p < 0.001 vs non-treated control.

Mentions: The morphometric analysis was carried out by measuring the longest neurite length per cell (Figure 3). After 24 and 48 h of treatment, the length of neurite obtain from cells treated with ECa 233 at the concentration of 0.1 μg/ml (23.02 ± 1.05 and 26.46 ± 0.87 μm) was not significantly different from those of non-treated cells (26.22 ± 1.19, 27.91 ± 1.01 μm). Whereas, neurite lengths exposed to ECa 233 at the concentrations of 1, 10 and 100 μg/ml were significantly enhanced in a time-dependent manner (35.58 ± 2.61, 37.39 ± 2.69 and 39.9 ± 2.73 μm at 24 h, and 47.57 ± 2.84, 44.03 ± 1.66 and 46.52 ± 2.71 μm at 48 h). Similar results were obtained when the cell were treated with BDNF 100 ng/ml (35.83 ± 2.17 and 54.38 ± 4.06 μm). Notably, the effect of ECa 233 at 1, 10 and 100 μg/ml was comparable to that of BDNF with no significant difference.


Neuritogenic effect of standardized extract of Centella asiatica ECa233 on human neuroblastoma cells.

Wanakhachornkrai O, Pongrakhananon V, Chunhacha P, Wanasuntronwong A, Vattanajun A, Tantisira B, Chanvorachote P, Tantisira MH - BMC Complement Altern Med (2013)

Morphometric analysis of IMR-32 cells affected by ECa 233. After cells cultured for 24 and 48 h in the presence of either ECa 233 (0.1, 1, 10 and 100 μg/ml) or BDNF (100 ng/ml). Morphometric analysis was carried out by measuring neurites from 60 living cells per treated condition, (n = 3). Average of the neurite length was significantly increased after being treated with ECa 233 (1, 10 and 100 μg/ml) or BDNF. The data presented as mean ± S.E. ** = p < 0.01, *** = p < 0.001 vs non-treated control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750251&req=5

Figure 3: Morphometric analysis of IMR-32 cells affected by ECa 233. After cells cultured for 24 and 48 h in the presence of either ECa 233 (0.1, 1, 10 and 100 μg/ml) or BDNF (100 ng/ml). Morphometric analysis was carried out by measuring neurites from 60 living cells per treated condition, (n = 3). Average of the neurite length was significantly increased after being treated with ECa 233 (1, 10 and 100 μg/ml) or BDNF. The data presented as mean ± S.E. ** = p < 0.01, *** = p < 0.001 vs non-treated control.
Mentions: The morphometric analysis was carried out by measuring the longest neurite length per cell (Figure 3). After 24 and 48 h of treatment, the length of neurite obtain from cells treated with ECa 233 at the concentration of 0.1 μg/ml (23.02 ± 1.05 and 26.46 ± 0.87 μm) was not significantly different from those of non-treated cells (26.22 ± 1.19, 27.91 ± 1.01 μm). Whereas, neurite lengths exposed to ECa 233 at the concentrations of 1, 10 and 100 μg/ml were significantly enhanced in a time-dependent manner (35.58 ± 2.61, 37.39 ± 2.69 and 39.9 ± 2.73 μm at 24 h, and 47.57 ± 2.84, 44.03 ± 1.66 and 46.52 ± 2.71 μm at 48 h). Similar results were obtained when the cell were treated with BDNF 100 ng/ml (35.83 ± 2.17 and 54.38 ± 4.06 μm). Notably, the effect of ECa 233 at 1, 10 and 100 μg/ml was comparable to that of BDNF with no significant difference.

Bottom Line: While ECa 233 had only limited effects on cell viability, it significantly enhanced neurite outgrowth of IMR-32 cells at the concentrations of 1-100 μg/ml.ERK1/2 and Akt signaling pathways seemed to account for the neurotrophic effect observed.In conjunction with in vivo neuroprotective effect of ECa 233 previously reported, the results obtained support further development of ECa 233 for clinical use in neuronal injury or neurodegenerative diseases.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Background: In order to gain insight into neuroprotective effects of ECa 233, a standardized extract of Centella asiatica, previously demonstrated in animal models of memory impairment induced by transient global ischemia or intracerebroventricular injection of β-amyloid, the effect of ECa 233 on neurite outgrowth of human IMR-32 neuroblastoma cell line was investigated.

Methods: Cells were seeded and incubated with various concentrations of ECa 233. Morphometric analysis was carried out by a measurement of the longest neurite growth of cells at 24 and 48 h. Contributing signaling pathways possibly involved were subsequently elucidated by western blot analysis.

Results: While ECa 233 had only limited effects on cell viability, it significantly enhanced neurite outgrowth of IMR-32 cells at the concentrations of 1-100 μg/ml. Western blot analysis revealed that ECa 233 significantly upregulated the level of activated ERK1/2 and Akt of the treated cells suggesting their involvement in the neuritogenic effect observed, which was subsequently verified by the finding that an addition of their respective inhibitors could reverse the effect of ECa 233 on these cells.

Conclusions: The present study clearly demonstrated neurite outgrowth promoting activity of ECa 233. ERK1/2 and Akt signaling pathways seemed to account for the neurotrophic effect observed. In conjunction with in vivo neuroprotective effect of ECa 233 previously reported, the results obtained support further development of ECa 233 for clinical use in neuronal injury or neurodegenerative diseases.

Show MeSH
Related in: MedlinePlus