Limits...
Neuroprotective effects of erucin against 6-hydroxydopamine-induced oxidative damage in a dopaminergic-like neuroblastoma cell line.

Tarozzi A, Morroni F, Bolondi C, Sita G, Hrelia P, Djemil A, Cantelli-Forti G - Int J Mol Sci (2012)

Bottom Line: The increase of GSH levels was associated with an increase in the resistance of SH-SY5Y cells to neuronal death, in terms of apoptosis, induced by 6-hydroxydopamine (6-OHDA).Last, the antiapoptotic and antioxidant effects of ER were abolished by buthionine sulfoximine, supporting the main role of GSH in the neuroprotective effects recorded by ER.These results suggest that ER may prevent the oxidative damage induced by 6-OHDA.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Alma Mater Studiorum-University of Bologna, via Irnerio 48, 40126 Bologna, Italy; E-Mails: fabiana.morroni@unibo.it (F.M.); cecilia.bolondi2@unibo.it (C.B.); giulia.sita2@unibo.it (G.S.); patrizia.hrelia@unibo.it (P.H.); alice.djemil@studio.unibo.it (A.D.); giorgio.cantelliforti@unibo.it (G.C.-F.).

ABSTRACT
Oxidative stress (OS) contributes to the cascade leading to the dysfunction or death of dopaminergic neurons during Parkinson's disease (PD). A strategy to prevent the OS of dopaminergic neurons may be the use of phytochemicals as inducers of endogenous antioxidants and phase 2 enzymes. In this study, we demonstrated that treatment of the dopaminergic-like neuroblastoma SH-SY5Y cell line with isothiocyanate erucin (ER), a compound of cruciferous vegetables, resulted in significant increases of both total glutathione (GSH) levels and total antioxidant capacity at the cytosolic level. The increase of GSH levels was associated with an increase in the resistance of SH-SY5Y cells to neuronal death, in terms of apoptosis, induced by 6-hydroxydopamine (6-OHDA). The pretreatment of SH-SY5Y cells with ER was also shown to prevent the redox status impairment, in terms of intracellular ROS and O(2) (•-) formation, and loss of mitochondrial membrane potential, early events that are initiators of the apoptotic process, induced by 6-OHDA. Last, the antiapoptotic and antioxidant effects of ER were abolished by buthionine sulfoximine, supporting the main role of GSH in the neuroprotective effects recorded by ER. These results suggest that ER may prevent the oxidative damage induced by 6-OHDA.

No MeSH data available.


Related in: MedlinePlus

Isothiocyanate erucin (ER) enhances the total glutathione (GSH) level and the total antioxidant capacity (TAC) of SH-SY5Y cells. (a) SH-SY5Y cells were incubated with various concentrations of ER for 24 h. At the end of the incubation, GSH levels were measured as described in the Experimental Section. The values are shown as mean ± SEM of four independent experiments. *p < 0.05, **p < 0.01 versus untreated cells at ANOVA with Dunnett post hoc test; (b) SH-SY5Y cells were treated with 5 μmol/L of ER for 24 h and cytosolic and membrane fractions were then separated as described in the Experimental Section. The cellular fractions were submitted to the ABTS•+ decolorization assay and the TAC of the fractions was expressed as μmol of trolox equivalent (TE) Antioxidant Activity per mg of protein (TEAA μmol/mg protein). The values are shown as mean ± SEM of three independent experiments. *p < 0.05 versus untreated cells at Student’s t-test.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC3472719&req=5

f1-ijms-13-10899: Isothiocyanate erucin (ER) enhances the total glutathione (GSH) level and the total antioxidant capacity (TAC) of SH-SY5Y cells. (a) SH-SY5Y cells were incubated with various concentrations of ER for 24 h. At the end of the incubation, GSH levels were measured as described in the Experimental Section. The values are shown as mean ± SEM of four independent experiments. *p < 0.05, **p < 0.01 versus untreated cells at ANOVA with Dunnett post hoc test; (b) SH-SY5Y cells were treated with 5 μmol/L of ER for 24 h and cytosolic and membrane fractions were then separated as described in the Experimental Section. The cellular fractions were submitted to the ABTS•+ decolorization assay and the TAC of the fractions was expressed as μmol of trolox equivalent (TE) Antioxidant Activity per mg of protein (TEAA μmol/mg protein). The values are shown as mean ± SEM of three independent experiments. *p < 0.05 versus untreated cells at Student’s t-test.

Mentions: Preliminary experiments showed that the treatment of SH-SY5Y cells with ER concentrations up to 5 μmol/L for 24 h did not affect intracellular redox state, MMP, cell growth and viability (data not shown). We first determined the ability of ER (1.25–5 μmol/L) to increase the total GSH levels in SH-SY5Y cells. Treatment of SH-SY5Y cells with 2.5 and 5 μmol/L of ER for 24 h resulted in significant increases (~35% and ~45%, respectively) in cellular GSH content (Figure 1a). We then evaluated whether the higher levels of GSH induced by ER effectively improves the TAC of SH-SY5Y cells at different subcellular levels, such as the membrane and cytosol. TAC was measured for its ability to quench the 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical cation (ABTS•+) and the results are expressed as trolox equivalent (TE) μmol/mg protein. The cytosolic, but not the membrane, fractions obtained from SH-SY5Y cells treated with 5 μmol/L of ER for 24 h showed significant increases of TAC compared to untreated cells (Figure 1b).


Neuroprotective effects of erucin against 6-hydroxydopamine-induced oxidative damage in a dopaminergic-like neuroblastoma cell line.

Tarozzi A, Morroni F, Bolondi C, Sita G, Hrelia P, Djemil A, Cantelli-Forti G - Int J Mol Sci (2012)

Isothiocyanate erucin (ER) enhances the total glutathione (GSH) level and the total antioxidant capacity (TAC) of SH-SY5Y cells. (a) SH-SY5Y cells were incubated with various concentrations of ER for 24 h. At the end of the incubation, GSH levels were measured as described in the Experimental Section. The values are shown as mean ± SEM of four independent experiments. *p < 0.05, **p < 0.01 versus untreated cells at ANOVA with Dunnett post hoc test; (b) SH-SY5Y cells were treated with 5 μmol/L of ER for 24 h and cytosolic and membrane fractions were then separated as described in the Experimental Section. The cellular fractions were submitted to the ABTS•+ decolorization assay and the TAC of the fractions was expressed as μmol of trolox equivalent (TE) Antioxidant Activity per mg of protein (TEAA μmol/mg protein). The values are shown as mean ± SEM of three independent experiments. *p < 0.05 versus untreated cells at Student’s t-test.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3472719&req=5

f1-ijms-13-10899: Isothiocyanate erucin (ER) enhances the total glutathione (GSH) level and the total antioxidant capacity (TAC) of SH-SY5Y cells. (a) SH-SY5Y cells were incubated with various concentrations of ER for 24 h. At the end of the incubation, GSH levels were measured as described in the Experimental Section. The values are shown as mean ± SEM of four independent experiments. *p < 0.05, **p < 0.01 versus untreated cells at ANOVA with Dunnett post hoc test; (b) SH-SY5Y cells were treated with 5 μmol/L of ER for 24 h and cytosolic and membrane fractions were then separated as described in the Experimental Section. The cellular fractions were submitted to the ABTS•+ decolorization assay and the TAC of the fractions was expressed as μmol of trolox equivalent (TE) Antioxidant Activity per mg of protein (TEAA μmol/mg protein). The values are shown as mean ± SEM of three independent experiments. *p < 0.05 versus untreated cells at Student’s t-test.
Mentions: Preliminary experiments showed that the treatment of SH-SY5Y cells with ER concentrations up to 5 μmol/L for 24 h did not affect intracellular redox state, MMP, cell growth and viability (data not shown). We first determined the ability of ER (1.25–5 μmol/L) to increase the total GSH levels in SH-SY5Y cells. Treatment of SH-SY5Y cells with 2.5 and 5 μmol/L of ER for 24 h resulted in significant increases (~35% and ~45%, respectively) in cellular GSH content (Figure 1a). We then evaluated whether the higher levels of GSH induced by ER effectively improves the TAC of SH-SY5Y cells at different subcellular levels, such as the membrane and cytosol. TAC was measured for its ability to quench the 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) radical cation (ABTS•+) and the results are expressed as trolox equivalent (TE) μmol/mg protein. The cytosolic, but not the membrane, fractions obtained from SH-SY5Y cells treated with 5 μmol/L of ER for 24 h showed significant increases of TAC compared to untreated cells (Figure 1b).

Bottom Line: The increase of GSH levels was associated with an increase in the resistance of SH-SY5Y cells to neuronal death, in terms of apoptosis, induced by 6-hydroxydopamine (6-OHDA).Last, the antiapoptotic and antioxidant effects of ER were abolished by buthionine sulfoximine, supporting the main role of GSH in the neuroprotective effects recorded by ER.These results suggest that ER may prevent the oxidative damage induced by 6-OHDA.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, Alma Mater Studiorum-University of Bologna, via Irnerio 48, 40126 Bologna, Italy; E-Mails: fabiana.morroni@unibo.it (F.M.); cecilia.bolondi2@unibo.it (C.B.); giulia.sita2@unibo.it (G.S.); patrizia.hrelia@unibo.it (P.H.); alice.djemil@studio.unibo.it (A.D.); giorgio.cantelliforti@unibo.it (G.C.-F.).

ABSTRACT
Oxidative stress (OS) contributes to the cascade leading to the dysfunction or death of dopaminergic neurons during Parkinson's disease (PD). A strategy to prevent the OS of dopaminergic neurons may be the use of phytochemicals as inducers of endogenous antioxidants and phase 2 enzymes. In this study, we demonstrated that treatment of the dopaminergic-like neuroblastoma SH-SY5Y cell line with isothiocyanate erucin (ER), a compound of cruciferous vegetables, resulted in significant increases of both total glutathione (GSH) levels and total antioxidant capacity at the cytosolic level. The increase of GSH levels was associated with an increase in the resistance of SH-SY5Y cells to neuronal death, in terms of apoptosis, induced by 6-hydroxydopamine (6-OHDA). The pretreatment of SH-SY5Y cells with ER was also shown to prevent the redox status impairment, in terms of intracellular ROS and O(2) (•-) formation, and loss of mitochondrial membrane potential, early events that are initiators of the apoptotic process, induced by 6-OHDA. Last, the antiapoptotic and antioxidant effects of ER were abolished by buthionine sulfoximine, supporting the main role of GSH in the neuroprotective effects recorded by ER. These results suggest that ER may prevent the oxidative damage induced by 6-OHDA.

No MeSH data available.


Related in: MedlinePlus