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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 inhibited 6-OHDA-induced nitric oxide (NO) over-production in PC12 cells. PC12 cells were pretreated with or without 250 μM L-NAME, 100 μM SNP and 25, 50, 100 and 200 μg/mL EBE for 12 hours, then exposed to1 mM 6-OHDA for another hour. (A-H) Intracellular NO was identified using fluorescent indicator, DAF-FM diacetate; (I) The NO fluorescent intensity was quantified by a multi-label counter. ++ P = 0.008 versus control group (without 6-OHDA treatment); * P = 0.044 versus OHDA group; ** P = 0.005 versus OHDA group. All experiments were repeated 3 times.
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Figure 7: EBE inhibited 6-OHDA-induced nitric oxide (NO) over-production in PC12 cells. PC12 cells were pretreated with or without 250 μM L-NAME, 100 μM SNP and 25, 50, 100 and 200 μg/mL EBE for 12 hours, then exposed to1 mM 6-OHDA for another hour. (A-H) Intracellular NO was identified using fluorescent indicator, DAF-FM diacetate; (I) The NO fluorescent intensity was quantified by a multi-label counter. ++ P = 0.008 versus control group (without 6-OHDA treatment); * P = 0.044 versus OHDA group; ** P = 0.005 versus OHDA group. All experiments were repeated 3 times.

Mentions: Figure 7 shows that 6-OHDA exposure led to a roughly 1.5-fold increase in NO production compared with the control group; similar elevation was also observed when SNP (sodium nitroprusside dehydrate, NO generator) was added. This increase in NO level was reduced in a concentration-dependent manner by pretreatment with EBE in a 25-200 μg/mL range for 12 hours (Figure 7E-H). Pretreatment with 250 μM L-NAME for 12 hours also significantly (P = 0.044) reduced this 6-OHDA-induced NO over-production. Pretreatment with higher concentrations (50, 100, 200 μg/ml) of EBE suppressed the elevated NO production more efficiently than L-NAME (Figure 7D). Furthermore, DAF-FM diacetate staining was photographed with a fluorescent microscope (Figure 7A-H). The expression of iNOS was up-regulated by 6-OHDA exposure in Western blot analysis and such up-regulation was prevented by pretreatment with EBE (Figure 8A).


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 inhibited 6-OHDA-induced nitric oxide (NO) over-production in PC12 cells. PC12 cells were pretreated with or without 250 μM L-NAME, 100 μM SNP and 25, 50, 100 and 200 μg/mL EBE for 12 hours, then exposed to1 mM 6-OHDA for another hour. (A-H) Intracellular NO was identified using fluorescent indicator, DAF-FM diacetate; (I) The NO fluorescent intensity was quantified by a multi-label counter. ++ P = 0.008 versus control group (without 6-OHDA treatment); * P = 0.044 versus OHDA group; ** P = 0.005 versus OHDA group. All experiments were repeated 3 times.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3108929&req=5

Figure 7: EBE inhibited 6-OHDA-induced nitric oxide (NO) over-production in PC12 cells. PC12 cells were pretreated with or without 250 μM L-NAME, 100 μM SNP and 25, 50, 100 and 200 μg/mL EBE for 12 hours, then exposed to1 mM 6-OHDA for another hour. (A-H) Intracellular NO was identified using fluorescent indicator, DAF-FM diacetate; (I) The NO fluorescent intensity was quantified by a multi-label counter. ++ P = 0.008 versus control group (without 6-OHDA treatment); * P = 0.044 versus OHDA group; ** P = 0.005 versus OHDA group. All experiments were repeated 3 times.
Mentions: Figure 7 shows that 6-OHDA exposure led to a roughly 1.5-fold increase in NO production compared with the control group; similar elevation was also observed when SNP (sodium nitroprusside dehydrate, NO generator) was added. This increase in NO level was reduced in a concentration-dependent manner by pretreatment with EBE in a 25-200 μg/mL range for 12 hours (Figure 7E-H). Pretreatment with 250 μM L-NAME for 12 hours also significantly (P = 0.044) reduced this 6-OHDA-induced NO over-production. Pretreatment with higher concentrations (50, 100, 200 μg/ml) of EBE suppressed the elevated NO production more efficiently than L-NAME (Figure 7D). Furthermore, DAF-FM diacetate staining was photographed with a fluorescent microscope (Figure 7A-H). The expression of iNOS was up-regulated by 6-OHDA exposure in Western blot analysis and such up-regulation was prevented by pretreatment with EBE (Figure 8A).

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