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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

EBE dose-dependently reduced PC12 cell death induced by 6-OHDA. PC12 cells were pretreated with or without EBE for 12 hours, pretreatment with 250 μM L-NAME for 12 hours served as positive control. The cells were exposed to 1 mM 6-OHDA for another 12 hours after pretreatment. (A) Cell viability was measured by MTT assay and results were expressed as percentage of control group (without 6-OHDA treatment). (B) Cell viability was measured as the percentage of released LDH. +++ P < 0.0001 versus control group (without 6-OHDA treatment); * P = 0.028 versus 6-OHDA group; *** P < 0.0001 versus 6-OHDA group. Each experiment was repeated 3 times.
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Figure 5: EBE dose-dependently reduced PC12 cell death induced by 6-OHDA. PC12 cells were pretreated with or without EBE for 12 hours, pretreatment with 250 μM L-NAME for 12 hours served as positive control. The cells were exposed to 1 mM 6-OHDA for another 12 hours after pretreatment. (A) Cell viability was measured by MTT assay and results were expressed as percentage of control group (without 6-OHDA treatment). (B) Cell viability was measured as the percentage of released LDH. +++ P < 0.0001 versus control group (without 6-OHDA treatment); * P = 0.028 versus 6-OHDA group; *** P < 0.0001 versus 6-OHDA group. Each experiment was repeated 3 times.

Mentions: The cell viability of PC12 cells exposed to 1 mM 6-OHDA for 12 hours was significantly decreased (46.2% ± 9.2%; P < 0.0001) compared with the control group (Figure 5A). Pretreatment with EBE of various concentrations (25, 50, 100 and 200 μg/ml) for 12 hours protected PC 12 cells against 6-OHDA-induced cellular damage in a dose-dependent manner. Compared with the control, the survival rates of the EBE treatment groups (25, 50, 100, 200 μg/ml) were 63.4% ± 7.3%, 77.1% ± 4.5%, 88.8% and 102.3% respectively. Toxicity was not observed when the cells were treated with 200 μg/mL alone. LDH is released from the cells following membrane damage, as a sign of cell death. Evaluation of LDH release revealed a significant increase after 6-OHDA exposure in PC12 cells while pretreatment with EBE suppressed the 6-OHDA-induced LDH release in a dose dependent manner (Figure 5B). L-NAME is an inhibitor of NOS and served as a positive control. Significant protective effects were found in the positive control groups with both MTT and LDH assays (Figures 5A and 5B).


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)

EBE dose-dependently reduced PC12 cell death induced by 6-OHDA. PC12 cells were pretreated with or without EBE for 12 hours, pretreatment with 250 μM L-NAME for 12 hours served as positive control. The cells were exposed to 1 mM 6-OHDA for another 12 hours after pretreatment. (A) Cell viability was measured by MTT assay and results were expressed as percentage of control group (without 6-OHDA treatment). (B) Cell viability was measured as the percentage of released LDH. +++ P < 0.0001 versus control group (without 6-OHDA treatment); * P = 0.028 versus 6-OHDA group; *** P < 0.0001 versus 6-OHDA group. Each experiment was repeated 3 times.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3108929&req=5

Figure 5: EBE dose-dependently reduced PC12 cell death induced by 6-OHDA. PC12 cells were pretreated with or without EBE for 12 hours, pretreatment with 250 μM L-NAME for 12 hours served as positive control. The cells were exposed to 1 mM 6-OHDA for another 12 hours after pretreatment. (A) Cell viability was measured by MTT assay and results were expressed as percentage of control group (without 6-OHDA treatment). (B) Cell viability was measured as the percentage of released LDH. +++ P < 0.0001 versus control group (without 6-OHDA treatment); * P = 0.028 versus 6-OHDA group; *** P < 0.0001 versus 6-OHDA group. Each experiment was repeated 3 times.
Mentions: The cell viability of PC12 cells exposed to 1 mM 6-OHDA for 12 hours was significantly decreased (46.2% ± 9.2%; P < 0.0001) compared with the control group (Figure 5A). Pretreatment with EBE of various concentrations (25, 50, 100 and 200 μg/ml) for 12 hours protected PC 12 cells against 6-OHDA-induced cellular damage in a dose-dependent manner. Compared with the control, the survival rates of the EBE treatment groups (25, 50, 100, 200 μg/ml) were 63.4% ± 7.3%, 77.1% ± 4.5%, 88.8% and 102.3% respectively. Toxicity was not observed when the cells were treated with 200 μg/mL alone. LDH is released from the cells following membrane damage, as a sign of cell death. Evaluation of LDH release revealed a significant increase after 6-OHDA exposure in PC12 cells while pretreatment with EBE suppressed the 6-OHDA-induced LDH release in a dose dependent manner (Figure 5B). L-NAME is an inhibitor of NOS and served as a positive control. Significant protective effects were found in the positive control groups with both MTT and LDH assays (Figures 5A and 5B).

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