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Eriocaulon buergerianum extract protects PC12 cells and neurons in zebrafish against 6-hydroxydopamine-induced damage.

Wang M, Zhang Z, Cheang LC, Lin Z, Lee SM - Chin Med (2011)

Bottom Line: EBE inhibited the 6-OHDA-induced decrease in total distance of movement in zebrafish.EBE exhibited significant neuroprotective activities in zebrafish, including recovery of dopaminergic neuron loss caused by 6-OHDA in a dose-dependent manner in vivo, inhibition of 6-OHDA-induced decrease of total distance in movement in zebrafish.The iNOS-NO pathway may be involved.

View Article: PubMed Central - HTML - PubMed

Affiliation: State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Av, Padre Tomás Pereira, Taipa, Macao, China. SimonLee@umac.mo.

ABSTRACT

Background: Ericaulon buergerianum (Gujingcao) is an ophthalmic, anti-inflammatory and antimicrobial Chinese medicinal herb. This study aims to investigate the neuroprotective effects of Ericaulon buergerianum ethanol extract (EBE) and to elucidate its underlying action mechanism.

Methods: The viability of dopaminergic (DA) neuron in zebrafish was examined by anti-tyrosine hydroxylase (TH) immunostaining. The locomotor activity of zebrafish was assessed with a digital video tracking system. The viability and cellular damage of the PC12 cells were determined by MTT and LDH assays respectively. The nuclear morphological changes in apoptotic cells were evaluated with DNA staining by Hoechst 33342 dye. Intracellular nitric oxide (NO) was quantified by DAF-FM diacetate staining. The expression of inducible nitric oxide synthase (iNOS) was determined by Western blot.

Results: EBE inhibited the 6-OHDA-induced decrease in total distance of movement in zebrafish. Pretreatments of EBE (25, 50, 100 and 200 μg/ml) increased the viability of 6-OHDA-damaged PC12 cells in a dose dependent manner. Protection against 6-OHDA-induced nuclear fragmentation and accumulation of apoptotic bodies was also observed in EBE pretreated cells. Anti-oxidative (inhibition of NO production and iNOS expression in PC12 cells in vitro) activities of EBE are related to its neuroprotective effects in 6-OHDA-induced DA neuron damage.

Conclusion: EBE exhibited significant neuroprotective activities in zebrafish, including recovery of dopaminergic neuron loss caused by 6-OHDA in a dose-dependent manner in vivo, inhibition of 6-OHDA-induced decrease of total distance in movement in zebrafish. The iNOS-NO pathway may be involved.

No MeSH data available.


Related in: MedlinePlus

Quantitative analysis of area of TH+ neuron in zebrafish brain. All data expressed as percentage of control group, each bar represents mean ± SD. +++ P < 0.0001 versus control group (without 6-OHDA treatment); * P = 0.046 versus 6-OHDA-treated group; *** P < 0.0001 versus 6-OHDA-treated group. All experiments were repeated 3 times.
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Figure 3: Quantitative analysis of area of TH+ neuron in zebrafish brain. All data expressed as percentage of control group, each bar represents mean ± SD. +++ P < 0.0001 versus control group (without 6-OHDA treatment); * P = 0.046 versus 6-OHDA-treated group; *** P < 0.0001 versus 6-OHDA-treated group. All experiments were repeated 3 times.

Mentions: All the tyrosine hydroxylase (TH)-positive neurons in zebrafish diencephalons are dopaminergic neurons [21]. Anti-TH immunostaining was used to compare the viability of dopaminergic neurons in zebrafish receiving different drug treatments (Figure 2). 2 dpf zebrafish were exposed to 250 μM 6-OHDA for 24 hours, dopaminergic neurons in the ventral diencephalic clusters (indicated by white bracket) were significantly reduced when compared to the control. Co-treatment with 6-OHDA and EBE for 24 hours recovered 6-OHDA-induced dopaminergic neuron loss in a dose-dependent manner (P = 0.046 at 25 μg/ml, *** P < 0.0001 at 25 and 50 μg/ml compared with 6-OHDA treatment alone) (Figure 3). Nomifensin (DAT inhibitor) and L-NAME were used as positive controls, both significantly reversed the loss of dopaminergic neurons caused by 6-OHDA (P < 0.0001).


Eriocaulon buergerianum extract protects PC12 cells and neurons in zebrafish against 6-hydroxydopamine-induced damage.

Wang M, Zhang Z, Cheang LC, Lin Z, Lee SM - Chin Med (2011)

Quantitative analysis of area of TH+ neuron in zebrafish brain. All data expressed as percentage of control group, each bar represents mean ± SD. +++ P < 0.0001 versus control group (without 6-OHDA treatment); * P = 0.046 versus 6-OHDA-treated group; *** P < 0.0001 versus 6-OHDA-treated group. All experiments were repeated 3 times.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3108929&req=5

Figure 3: Quantitative analysis of area of TH+ neuron in zebrafish brain. All data expressed as percentage of control group, each bar represents mean ± SD. +++ P < 0.0001 versus control group (without 6-OHDA treatment); * P = 0.046 versus 6-OHDA-treated group; *** P < 0.0001 versus 6-OHDA-treated group. All experiments were repeated 3 times.
Mentions: All the tyrosine hydroxylase (TH)-positive neurons in zebrafish diencephalons are dopaminergic neurons [21]. Anti-TH immunostaining was used to compare the viability of dopaminergic neurons in zebrafish receiving different drug treatments (Figure 2). 2 dpf zebrafish were exposed to 250 μM 6-OHDA for 24 hours, dopaminergic neurons in the ventral diencephalic clusters (indicated by white bracket) were significantly reduced when compared to the control. Co-treatment with 6-OHDA and EBE for 24 hours recovered 6-OHDA-induced dopaminergic neuron loss in a dose-dependent manner (P = 0.046 at 25 μg/ml, *** P < 0.0001 at 25 and 50 μg/ml compared with 6-OHDA treatment alone) (Figure 3). Nomifensin (DAT inhibitor) and L-NAME were used as positive controls, both significantly reversed the loss of dopaminergic neurons caused by 6-OHDA (P < 0.0001).

Bottom Line: EBE inhibited the 6-OHDA-induced decrease in total distance of movement in zebrafish.EBE exhibited significant neuroprotective activities in zebrafish, including recovery of dopaminergic neuron loss caused by 6-OHDA in a dose-dependent manner in vivo, inhibition of 6-OHDA-induced decrease of total distance in movement in zebrafish.The iNOS-NO pathway may be involved.

View Article: PubMed Central - HTML - PubMed

Affiliation: State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Av, Padre Tomás Pereira, Taipa, Macao, China. SimonLee@umac.mo.

ABSTRACT

Background: Ericaulon buergerianum (Gujingcao) is an ophthalmic, anti-inflammatory and antimicrobial Chinese medicinal herb. This study aims to investigate the neuroprotective effects of Ericaulon buergerianum ethanol extract (EBE) and to elucidate its underlying action mechanism.

Methods: The viability of dopaminergic (DA) neuron in zebrafish was examined by anti-tyrosine hydroxylase (TH) immunostaining. The locomotor activity of zebrafish was assessed with a digital video tracking system. The viability and cellular damage of the PC12 cells were determined by MTT and LDH assays respectively. The nuclear morphological changes in apoptotic cells were evaluated with DNA staining by Hoechst 33342 dye. Intracellular nitric oxide (NO) was quantified by DAF-FM diacetate staining. The expression of inducible nitric oxide synthase (iNOS) was determined by Western blot.

Results: EBE inhibited the 6-OHDA-induced decrease in total distance of movement in zebrafish. Pretreatments of EBE (25, 50, 100 and 200 μg/ml) increased the viability of 6-OHDA-damaged PC12 cells in a dose dependent manner. Protection against 6-OHDA-induced nuclear fragmentation and accumulation of apoptotic bodies was also observed in EBE pretreated cells. Anti-oxidative (inhibition of NO production and iNOS expression in PC12 cells in vitro) activities of EBE are related to its neuroprotective effects in 6-OHDA-induced DA neuron damage.

Conclusion: EBE exhibited significant neuroprotective activities in zebrafish, including recovery of dopaminergic neuron loss caused by 6-OHDA in a dose-dependent manner in vivo, inhibition of 6-OHDA-induced decrease of total distance in movement in zebrafish. The iNOS-NO pathway may be involved.

No MeSH data available.


Related in: MedlinePlus