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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 reduced apoptosis induced by 6-OHDA in PC12 cells. Cells were stained with DNA-binding fluorescent dye Hoechst 33342. (A) Control: untreated group; (B) 6-OHDA-treated group (1 mM, 8 hours): chromatin condensation and DNA fragmentation were indicated by the white arrows; (C-F) EBE-pretreated groups (25, 50, 100 and 200 μg/mL respectively, 12 hours), followed by 6-OHDA exposure (1 mM, 8 hours): less apoptotic bodies were identified, colony reduction and cell shrinkage induced by 6-OHDA were also reversed.
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Figure 6: EBE reduced apoptosis induced by 6-OHDA in PC12 cells. Cells were stained with DNA-binding fluorescent dye Hoechst 33342. (A) Control: untreated group; (B) 6-OHDA-treated group (1 mM, 8 hours): chromatin condensation and DNA fragmentation were indicated by the white arrows; (C-F) EBE-pretreated groups (25, 50, 100 and 200 μg/mL respectively, 12 hours), followed by 6-OHDA exposure (1 mM, 8 hours): less apoptotic bodies were identified, colony reduction and cell shrinkage induced by 6-OHDA were also reversed.

Mentions: Apoptosis is morphologically characterized by cell shrinkage, chromatin condensation and nuclear fragmentation. To identify whether EBE reverses 6-OHDA-induced PC12 cell apoptosis, we used DNA staining with Hoechst 33342 to evaluate nuclear condensation. Normal untreated cells appeared circle or elliptical where no condensation of the nucleus was observable (Figure 6A). In contrast, bright condensed dots known as apoptotic bodies (indicated by arrows in Figure 6B) were clearly identified after exposure to 1 mM 6-OHDA for eight hours. Apoptotic bodies are generated when chromatin fragments are packaged in apoptotic cells, and are commonly accepted as a marker of apoptosis. Reductions in colony density and cell size were also observable when treated with 6-OHDA. These changes in nuclear characteristics of apoptosis were inhibited when the cells were pretreated with EBE of different concentrations (25, 50, 100, 200 μg/ml) (Figures 6C-F).


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 reduced apoptosis induced by 6-OHDA in PC12 cells. Cells were stained with DNA-binding fluorescent dye Hoechst 33342. (A) Control: untreated group; (B) 6-OHDA-treated group (1 mM, 8 hours): chromatin condensation and DNA fragmentation were indicated by the white arrows; (C-F) EBE-pretreated groups (25, 50, 100 and 200 μg/mL respectively, 12 hours), followed by 6-OHDA exposure (1 mM, 8 hours): less apoptotic bodies were identified, colony reduction and cell shrinkage induced by 6-OHDA were also reversed.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3108929&req=5

Figure 6: EBE reduced apoptosis induced by 6-OHDA in PC12 cells. Cells were stained with DNA-binding fluorescent dye Hoechst 33342. (A) Control: untreated group; (B) 6-OHDA-treated group (1 mM, 8 hours): chromatin condensation and DNA fragmentation were indicated by the white arrows; (C-F) EBE-pretreated groups (25, 50, 100 and 200 μg/mL respectively, 12 hours), followed by 6-OHDA exposure (1 mM, 8 hours): less apoptotic bodies were identified, colony reduction and cell shrinkage induced by 6-OHDA were also reversed.
Mentions: Apoptosis is morphologically characterized by cell shrinkage, chromatin condensation and nuclear fragmentation. To identify whether EBE reverses 6-OHDA-induced PC12 cell apoptosis, we used DNA staining with Hoechst 33342 to evaluate nuclear condensation. Normal untreated cells appeared circle or elliptical where no condensation of the nucleus was observable (Figure 6A). In contrast, bright condensed dots known as apoptotic bodies (indicated by arrows in Figure 6B) were clearly identified after exposure to 1 mM 6-OHDA for eight hours. Apoptotic bodies are generated when chromatin fragments are packaged in apoptotic cells, and are commonly accepted as a marker of apoptosis. Reductions in colony density and cell size were also observable when treated with 6-OHDA. These changes in nuclear characteristics of apoptosis were inhibited when the cells were pretreated with EBE of different concentrations (25, 50, 100, 200 μg/ml) (Figures 6C-F).

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