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Neurite outgrowth stimulatory effects of culinary-medicinal mushrooms and their toxicity assessment using differentiating Neuro-2a and embryonic fibroblast BALB/3T3.

Phan CW, David P, Naidu M, Wong KH, Sabaratnam V - BMC Complement Altern Med (2013)

Bottom Line: However, little toxicological information is available regarding their safety.The IC50 values obtained from tetrazolium (MTT), neutral red uptake (NRU) and lactate dehydrogenase (LDH) release assays showed no toxic effects following 24 h exposure of N2a and 3T3 cells to mushroom extracts.Our results indicate that G. lucidum, L. rhinocerotis, P. giganteus, G. frondosa and C. militaris may be developed as safe and healthy dietary supplements for brain and cognitive health.

View Article: PubMed Central - HTML - PubMed

Affiliation: Mushroom Research Centre, Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia. rosiepamela@um.edu.my.

ABSTRACT

Background: Mushrooms are not only regarded as gourmet cuisine but also as therapeutic agent to promote cognition health. However, little toxicological information is available regarding their safety. Therefore, the aim of this study was to screen selected ethno-pharmacologically important mushrooms for stimulatory effects on neurite outgrowth and to test for any cytotoxicity.

Methods: The stimulatory effect of mushrooms on neurite outgrowth was assessed in differentiating mouse neuroblastoma (N2a) cells. Neurite length was measured using Image-Pro Insight processor system. Neuritogenesis activity was further validated by fluorescence immunocytochemical staining of neurofilaments. In vitro cytotoxicity was investigated by using mouse embryonic fibroblast (BALB/3T3) and N2a cells for any embryo- and neuro-toxic effects; respectively.

Results: Aqueous extracts of Ganoderma lucidum, Lignosus rhinocerotis, Pleurotus giganteus and Grifola frondosa; as well as an ethanol extract of Cordyceps militaris significantly (p < 0.05) promoted the neurite outgrowth in N2a cells by 38.4 ± 4.2%, 38.1 ± 2.6%, 33.4 ± 4.6%, 33.7 ± 1.5%, and 35.8 ± 3.4%; respectively. The IC50 values obtained from tetrazolium (MTT), neutral red uptake (NRU) and lactate dehydrogenase (LDH) release assays showed no toxic effects following 24 h exposure of N2a and 3T3 cells to mushroom extracts.

Conclusion: Our results indicate that G. lucidum, L. rhinocerotis, P. giganteus, G. frondosa and C. militaris may be developed as safe and healthy dietary supplements for brain and cognitive health.

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Related in: MedlinePlus

The mean neurite length of N2a treated with different mushroom extracts at 20 μg/ml. The results shown represent the mean ± SD; n = 3. Means not sharing a common letter were significantly different at p < 0.05. S = sclerotium, aq = aqueous, EtOH = ethanol.
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Figure 3: The mean neurite length of N2a treated with different mushroom extracts at 20 μg/ml. The results shown represent the mean ± SD; n = 3. Means not sharing a common letter were significantly different at p < 0.05. S = sclerotium, aq = aqueous, EtOH = ethanol.

Mentions: The mean diameter of N2a cell body was found to be 19.45 ± 0.72 μm (Figure 3). To qualify as a “neurite”, the axon-like extension needs to be double or more than the cell body length of N2a. The average neurite length of NGF-stimulated cells was 78.58 ± 18.6 μm, which is approximately 4-time longer than the cell body. Cells treated with aqueous extract of G. lucidum were found to develop the longest mean neurite length i.e. 121.51 ± 28.6 μm (6.25-time longer than cell body), followed by aqueous extract of P. giganteus which recorded mean neurite length of 116.72 ± 29.5 μm (5.99-time longer than cell body). Figure 4 shows the morphology of differentiating N2a cells with neurites after 48 h of treatment with 50 ng/ml NGF (a) and 20 μg/ml of aqueous extracts of G. lucidum (b), L. rhinocerotis (c), P. giganteus (d) and G. frondosa (e); as well as ethanol extract of C. militaris (f).


Neurite outgrowth stimulatory effects of culinary-medicinal mushrooms and their toxicity assessment using differentiating Neuro-2a and embryonic fibroblast BALB/3T3.

Phan CW, David P, Naidu M, Wong KH, Sabaratnam V - BMC Complement Altern Med (2013)

The mean neurite length of N2a treated with different mushroom extracts at 20 μg/ml. The results shown represent the mean ± SD; n = 3. Means not sharing a common letter were significantly different at p < 0.05. S = sclerotium, aq = aqueous, EtOH = ethanol.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3852280&req=5

Figure 3: The mean neurite length of N2a treated with different mushroom extracts at 20 μg/ml. The results shown represent the mean ± SD; n = 3. Means not sharing a common letter were significantly different at p < 0.05. S = sclerotium, aq = aqueous, EtOH = ethanol.
Mentions: The mean diameter of N2a cell body was found to be 19.45 ± 0.72 μm (Figure 3). To qualify as a “neurite”, the axon-like extension needs to be double or more than the cell body length of N2a. The average neurite length of NGF-stimulated cells was 78.58 ± 18.6 μm, which is approximately 4-time longer than the cell body. Cells treated with aqueous extract of G. lucidum were found to develop the longest mean neurite length i.e. 121.51 ± 28.6 μm (6.25-time longer than cell body), followed by aqueous extract of P. giganteus which recorded mean neurite length of 116.72 ± 29.5 μm (5.99-time longer than cell body). Figure 4 shows the morphology of differentiating N2a cells with neurites after 48 h of treatment with 50 ng/ml NGF (a) and 20 μg/ml of aqueous extracts of G. lucidum (b), L. rhinocerotis (c), P. giganteus (d) and G. frondosa (e); as well as ethanol extract of C. militaris (f).

Bottom Line: However, little toxicological information is available regarding their safety.The IC50 values obtained from tetrazolium (MTT), neutral red uptake (NRU) and lactate dehydrogenase (LDH) release assays showed no toxic effects following 24 h exposure of N2a and 3T3 cells to mushroom extracts.Our results indicate that G. lucidum, L. rhinocerotis, P. giganteus, G. frondosa and C. militaris may be developed as safe and healthy dietary supplements for brain and cognitive health.

View Article: PubMed Central - HTML - PubMed

Affiliation: Mushroom Research Centre, Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia. rosiepamela@um.edu.my.

ABSTRACT

Background: Mushrooms are not only regarded as gourmet cuisine but also as therapeutic agent to promote cognition health. However, little toxicological information is available regarding their safety. Therefore, the aim of this study was to screen selected ethno-pharmacologically important mushrooms for stimulatory effects on neurite outgrowth and to test for any cytotoxicity.

Methods: The stimulatory effect of mushrooms on neurite outgrowth was assessed in differentiating mouse neuroblastoma (N2a) cells. Neurite length was measured using Image-Pro Insight processor system. Neuritogenesis activity was further validated by fluorescence immunocytochemical staining of neurofilaments. In vitro cytotoxicity was investigated by using mouse embryonic fibroblast (BALB/3T3) and N2a cells for any embryo- and neuro-toxic effects; respectively.

Results: Aqueous extracts of Ganoderma lucidum, Lignosus rhinocerotis, Pleurotus giganteus and Grifola frondosa; as well as an ethanol extract of Cordyceps militaris significantly (p < 0.05) promoted the neurite outgrowth in N2a cells by 38.4 ± 4.2%, 38.1 ± 2.6%, 33.4 ± 4.6%, 33.7 ± 1.5%, and 35.8 ± 3.4%; respectively. The IC50 values obtained from tetrazolium (MTT), neutral red uptake (NRU) and lactate dehydrogenase (LDH) release assays showed no toxic effects following 24 h exposure of N2a and 3T3 cells to mushroom extracts.

Conclusion: Our results indicate that G. lucidum, L. rhinocerotis, P. giganteus, G. frondosa and C. militaris may be developed as safe and healthy dietary supplements for brain and cognitive health.

Show MeSH
Related in: MedlinePlus