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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

Protective effect of broccoli sprouts juices from Aβ25–35-induced GSH depletion. 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 cell lysate was prepared for the determination of intracellular GSH levels by RP-HPLC analysis, using the fluorescence signal of the derivatized product with OPA. The concentration of GSH in the samples, normalized by the number of cells, was expressed as a percentage compared to the control. Data are represented as mean ± SEM (n = 3–5) aP < 0.05, bP < 0.01, and cP < 0.001 versus control; dP < 0.05 and eP < 0.01 versus Aβ25–35; fP < 0.05 versus (A + Aβ).
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Related In: Results  -  Collection


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fig5: Protective effect of broccoli sprouts juices from Aβ25–35-induced GSH depletion. 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 cell lysate was prepared for the determination of intracellular GSH levels by RP-HPLC analysis, using the fluorescence signal of the derivatized product with OPA. The concentration of GSH in the samples, normalized by the number of cells, was expressed as a percentage compared to the control. Data are represented as mean ± SEM (n = 3–5) aP < 0.05, bP < 0.01, and cP < 0.001 versus control; dP < 0.05 and eP < 0.01 versus Aβ25–35; fP < 0.05 versus (A + Aβ).

Mentions: It is well known that Aβ can induce oxidative stress via several different mechanisms thus triggering cell death [38]. To further verify whether Aβ25–35 treatment induces oxidative stress in our model, we determined the intracellular content of GSH, one of the major antioxidant defence systems in the cell. As shown in Figure 5, the treatment of SH-SY5Y cells with 25 μM Aβ25–35 up to 72 h was responsible for a rapid and significant reduction of intracellular GSH levels compared to the control group, respectively, of ~65% in the first 7 h of incubation, 81% after 24 h, and finally 55% after 48 h, up to 72 h of exposure. Both juice A and juice B, also determined a fast and significant decrease in the intracellular GSH levels compared to control cells, with a minimum achieved within 4 h of treatment (40% and 23% for juice A and juice B, resp.). On the other hand, after 7 h of incubation, and even more clearly within 24 h of exposure to the juices, a cellular response was observed that led to the progressive recovery of GSH levels comparable to those of the control. Notably, treatment for 48 h with the juices induced a substantial increase of the cellular content of GSH, 196% for juice A (P < 0.05) and 255% for juice B (P < 0.05) compared to control, which returned to basal levels within 72 h. The treatment of SH-SY5Y cells with 25 μM Aβ25–35 in the presence of 10 μL/mL of juice A or juice B showed a synergic effect on intracellular GSH depletion, reaching a minimum at 4 h of incubation (~20%, compared to the controls) and a recovery after 7 h of cotreatment. Within 48 h of exposure to Aβ25–35 and the juices, the intracellular GSH content recovered to the level of the control group in the presence of juice A (97%) and even increased up to 144% with the juice B (P < 0.05 versus Aβ25–35). In prolonged incubations up to 72 h, a protective effect on Aβ25–35-mediated depletion of intracellular GSH content was still observed in the presence of juice A (83% versus 55% for Aβ25–35, P < 0.05), while for the juice B such capacity was not appreciable (56% versus 55% for Aβ25–35, P > 0.05).


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)

Protective effect of broccoli sprouts juices from Aβ25–35-induced GSH depletion. 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 cell lysate was prepared for the determination of intracellular GSH levels by RP-HPLC analysis, using the fluorescence signal of the derivatized product with OPA. The concentration of GSH in the samples, normalized by the number of cells, was expressed as a percentage compared to the control. Data are represented as mean ± SEM (n = 3–5) aP < 0.05, bP < 0.01, and cP < 0.001 versus control; dP < 0.05 and eP < 0.01 versus Aβ25–35; fP < 0.05 versus (A + Aβ).
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig5: Protective effect of broccoli sprouts juices from Aβ25–35-induced GSH depletion. 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 cell lysate was prepared for the determination of intracellular GSH levels by RP-HPLC analysis, using the fluorescence signal of the derivatized product with OPA. The concentration of GSH in the samples, normalized by the number of cells, was expressed as a percentage compared to the control. Data are represented as mean ± SEM (n = 3–5) aP < 0.05, bP < 0.01, and cP < 0.001 versus control; dP < 0.05 and eP < 0.01 versus Aβ25–35; fP < 0.05 versus (A + Aβ).
Mentions: It is well known that Aβ can induce oxidative stress via several different mechanisms thus triggering cell death [38]. To further verify whether Aβ25–35 treatment induces oxidative stress in our model, we determined the intracellular content of GSH, one of the major antioxidant defence systems in the cell. As shown in Figure 5, the treatment of SH-SY5Y cells with 25 μM Aβ25–35 up to 72 h was responsible for a rapid and significant reduction of intracellular GSH levels compared to the control group, respectively, of ~65% in the first 7 h of incubation, 81% after 24 h, and finally 55% after 48 h, up to 72 h of exposure. Both juice A and juice B, also determined a fast and significant decrease in the intracellular GSH levels compared to control cells, with a minimum achieved within 4 h of treatment (40% and 23% for juice A and juice B, resp.). On the other hand, after 7 h of incubation, and even more clearly within 24 h of exposure to the juices, a cellular response was observed that led to the progressive recovery of GSH levels comparable to those of the control. Notably, treatment for 48 h with the juices induced a substantial increase of the cellular content of GSH, 196% for juice A (P < 0.05) and 255% for juice B (P < 0.05) compared to control, which returned to basal levels within 72 h. The treatment of SH-SY5Y cells with 25 μM Aβ25–35 in the presence of 10 μL/mL of juice A or juice B showed a synergic effect on intracellular GSH depletion, reaching a minimum at 4 h of incubation (~20%, compared to the controls) and a recovery after 7 h of cotreatment. Within 48 h of exposure to Aβ25–35 and the juices, the intracellular GSH content recovered to the level of the control group in the presence of juice A (97%) and even increased up to 144% with the juice B (P < 0.05 versus Aβ25–35). In prolonged incubations up to 72 h, a protective effect on Aβ25–35-mediated depletion of intracellular GSH content was still observed in the presence of juice A (83% versus 55% for Aβ25–35, P < 0.05), while for the juice B such capacity was not appreciable (56% versus 55% for Aβ25–35, P > 0.05).

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