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Neuroprotective Effect of Brassica oleracea Sprouts Crude Juice in a Cellular Model of Alzheimer's Disease.

Masci A, Mattioli R, Costantino P, Baima S, Morelli G, Punzi P, Giordano C, Pinto A, Donini LM, d'Erme M, Mosca L - Oxid Med Cell Longev (2015)

Bottom Line: In the present study, we analyzed the protective effect on SH-SY5Y cells treated with the fragment Aβ 25-35 by two crude juices of broccoli sprouts containing different amounts of phenolic compounds as a result of different growth conditions.Furthermore, the juices upregulated the intracellular glutathione content and mRNA levels or activity of antioxidant enzymes such as heme oxygenase-1, thioredoxin, thioredoxin reductase, and quinone oxidoreductase 1 via activation of NF-E2-related factor 2 (Nrf2).Although the effects of the two juices were similar, the juice enriched in phenolic compounds showed a greater efficacy in inducing the activation of the Nrf2 signalling pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Experimental Medicine-Medical Physiopathology, Food Science and Endocrinology Section, Sapienza University, Rome, Italy.

ABSTRACT

Unlabelled: β-Amyloid peptide (Aβ) aberrant production and aggregation are major factors implicated in the pathogenesis of Alzheimer's disease (AD), causing neuronal death via oxidative stress. Several studies have highlighted the importance of polyphenolic antioxidant compounds in the treatment of AD, but complex food matrices, characterized by a different relative content of these phytochemicals, have been neglected. In the present study, we analyzed the protective effect on SH-SY5Y cells treated with the fragment Aβ 25-35 by two crude juices of broccoli sprouts containing different amounts of phenolic compounds as a result of different growth conditions. Both juices protected against Aβ-induced cytotoxicity and apoptotic cell death as evidenced by cell viability, nuclear chromatin condensation, and apoptotic body formation measurements. These effects were mediated by the modulation of the mitochondrial function and of the HSP70 gene transcription and expression. Furthermore, the juices upregulated the intracellular glutathione content and mRNA levels or activity of antioxidant enzymes such as heme oxygenase-1, thioredoxin, thioredoxin reductase, and

Nad(p)h: quinone oxidoreductase 1 via activation of NF-E2-related factor 2 (Nrf2). Although the effects of the two juices were similar, the juice enriched in phenolic compounds showed a greater efficacy in inducing the activation of the Nrf2 signalling pathway.

No MeSH data available.


Related in: MedlinePlus

Broccoli sprouts juices protect SH-SY5Y cells from Aβ25–35-mediated apoptosis through upregulation of HSP70 gene expression. (a) SH-SY5Y cells were incubated with 25 μM Aβ25–35 in the presence or in the absence of 10 μL/mL of juice A or juice B for 1, 4, 7, 24, 48, and 72 h. After treatment total RNA was extracted and used for qRT-PCR determination of HSP70 gene induction. The RSP27A housekeeping gene was used as a control for normalization of the data. The relative amounts of gene expression were calculated by the comparative threshold cycle (ΔΔCT) method. The intracellular levels of mRNA are expressed as fold induction compared to control cells at 0 h of incubation. (b) Cell lysates obtained after 7 h of treatment were subjected to western blot analysis with anti-Hsp70 antidoby. Actin levels were compared to ensure equal amount of protein loading. The intracellular Hsp70 protein levels in the samples were expressed as a percentage compared to the control. Data are represented as mean ± SEM (n = 3), aP < 0.05 and bP < 0.01 versus control; cP < 0.05 and dP < 0.01 versus Aβ25–35.
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fig4: Broccoli sprouts juices protect SH-SY5Y cells from Aβ25–35-mediated apoptosis through upregulation of HSP70 gene expression. (a) SH-SY5Y cells were incubated with 25 μM Aβ25–35 in the presence or in the absence of 10 μL/mL of juice A or juice B for 1, 4, 7, 24, 48, and 72 h. After treatment total RNA was extracted and used for qRT-PCR determination of HSP70 gene induction. The RSP27A housekeeping gene was used as a control for normalization of the data. The relative amounts of gene expression were calculated by the comparative threshold cycle (ΔΔCT) method. The intracellular levels of mRNA are expressed as fold induction compared to control cells at 0 h of incubation. (b) Cell lysates obtained after 7 h of treatment were subjected to western blot analysis with anti-Hsp70 antidoby. Actin levels were compared to ensure equal amount of protein loading. The intracellular Hsp70 protein levels in the samples were expressed as a percentage compared to the control. Data are represented as mean ± SEM (n = 3), aP < 0.05 and bP < 0.01 versus control; cP < 0.05 and dP < 0.01 versus Aβ25–35.

Mentions: To elucidate the molecular mechanisms underlying the antiapoptotic effects of broccoli sprouts juices in SH-SY5Y cells, expression of the HSP70 gene was examined. The heat shock protein Hsp70 plays a key role in the cellular response for the protection from oxidative stress [34–36]. The determination by qRT-PCR of Hsp70 mRNA levels in SH-SY5Y cells (Figure 4(a)) showed no significant changes upon exposure to 25 μM Aβ25–35. Conversely, both broccoli sprouts juices, either in the absence or in the presence of Aβ25–35, induced an increase of HSP70 gene expression starting from 4 h of incubation and with a peak at 7 h (3.5–4 times versus control and Aβ25–35, P < 0.01), still persistent up to 24 h.


Neuroprotective Effect of Brassica oleracea Sprouts Crude Juice in a Cellular Model of Alzheimer's Disease.

Masci A, Mattioli R, Costantino P, Baima S, Morelli G, Punzi P, Giordano C, Pinto A, Donini LM, d'Erme M, Mosca L - Oxid Med Cell Longev (2015)

Broccoli sprouts juices protect SH-SY5Y cells from Aβ25–35-mediated apoptosis through upregulation of HSP70 gene expression. (a) SH-SY5Y cells were incubated with 25 μM Aβ25–35 in the presence or in the absence of 10 μL/mL of juice A or juice B for 1, 4, 7, 24, 48, and 72 h. After treatment total RNA was extracted and used for qRT-PCR determination of HSP70 gene induction. The RSP27A housekeeping gene was used as a control for normalization of the data. The relative amounts of gene expression were calculated by the comparative threshold cycle (ΔΔCT) method. The intracellular levels of mRNA are expressed as fold induction compared to control cells at 0 h of incubation. (b) Cell lysates obtained after 7 h of treatment were subjected to western blot analysis with anti-Hsp70 antidoby. Actin levels were compared to ensure equal amount of protein loading. The intracellular Hsp70 protein levels in the samples were expressed as a percentage compared to the control. Data are represented as mean ± SEM (n = 3), aP < 0.05 and bP < 0.01 versus control; cP < 0.05 and dP < 0.01 versus Aβ25–35.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4477226&req=5

fig4: Broccoli sprouts juices protect SH-SY5Y cells from Aβ25–35-mediated apoptosis through upregulation of HSP70 gene expression. (a) SH-SY5Y cells were incubated with 25 μM Aβ25–35 in the presence or in the absence of 10 μL/mL of juice A or juice B for 1, 4, 7, 24, 48, and 72 h. After treatment total RNA was extracted and used for qRT-PCR determination of HSP70 gene induction. The RSP27A housekeeping gene was used as a control for normalization of the data. The relative amounts of gene expression were calculated by the comparative threshold cycle (ΔΔCT) method. The intracellular levels of mRNA are expressed as fold induction compared to control cells at 0 h of incubation. (b) Cell lysates obtained after 7 h of treatment were subjected to western blot analysis with anti-Hsp70 antidoby. Actin levels were compared to ensure equal amount of protein loading. The intracellular Hsp70 protein levels in the samples were expressed as a percentage compared to the control. Data are represented as mean ± SEM (n = 3), aP < 0.05 and bP < 0.01 versus control; cP < 0.05 and dP < 0.01 versus Aβ25–35.
Mentions: To elucidate the molecular mechanisms underlying the antiapoptotic effects of broccoli sprouts juices in SH-SY5Y cells, expression of the HSP70 gene was examined. The heat shock protein Hsp70 plays a key role in the cellular response for the protection from oxidative stress [34–36]. The determination by qRT-PCR of Hsp70 mRNA levels in SH-SY5Y cells (Figure 4(a)) showed no significant changes upon exposure to 25 μM Aβ25–35. Conversely, both broccoli sprouts juices, either in the absence or in the presence of Aβ25–35, induced an increase of HSP70 gene expression starting from 4 h of incubation and with a peak at 7 h (3.5–4 times versus control and Aβ25–35, P < 0.01), still persistent up to 24 h.

Bottom Line: In the present study, we analyzed the protective effect on SH-SY5Y cells treated with the fragment Aβ 25-35 by two crude juices of broccoli sprouts containing different amounts of phenolic compounds as a result of different growth conditions.Furthermore, the juices upregulated the intracellular glutathione content and mRNA levels or activity of antioxidant enzymes such as heme oxygenase-1, thioredoxin, thioredoxin reductase, and quinone oxidoreductase 1 via activation of NF-E2-related factor 2 (Nrf2).Although the effects of the two juices were similar, the juice enriched in phenolic compounds showed a greater efficacy in inducing the activation of the Nrf2 signalling pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Experimental Medicine-Medical Physiopathology, Food Science and Endocrinology Section, Sapienza University, Rome, Italy.

ABSTRACT

Unlabelled: β-Amyloid peptide (Aβ) aberrant production and aggregation are major factors implicated in the pathogenesis of Alzheimer's disease (AD), causing neuronal death via oxidative stress. Several studies have highlighted the importance of polyphenolic antioxidant compounds in the treatment of AD, but complex food matrices, characterized by a different relative content of these phytochemicals, have been neglected. In the present study, we analyzed the protective effect on SH-SY5Y cells treated with the fragment Aβ 25-35 by two crude juices of broccoli sprouts containing different amounts of phenolic compounds as a result of different growth conditions. Both juices protected against Aβ-induced cytotoxicity and apoptotic cell death as evidenced by cell viability, nuclear chromatin condensation, and apoptotic body formation measurements. These effects were mediated by the modulation of the mitochondrial function and of the HSP70 gene transcription and expression. Furthermore, the juices upregulated the intracellular glutathione content and mRNA levels or activity of antioxidant enzymes such as heme oxygenase-1, thioredoxin, thioredoxin reductase, and

Nad(p)h: quinone oxidoreductase 1 via activation of NF-E2-related factor 2 (Nrf2). Although the effects of the two juices were similar, the juice enriched in phenolic compounds showed a greater efficacy in inducing the activation of the Nrf2 signalling pathway.

No MeSH data available.


Related in: MedlinePlus