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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 upregulate the cellular antioxidant defence capacity. 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 gene expression of (a) HO-1; (b) TRX; (c) TrxR. 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. (d) Total cell lysates were subjected to spectrophotometric assay of NQO1 activity which was expressed as a percentage compared to the control. Data are represented as mean ± SEM (n = 3), aP < 0.05, bP < 0.01, and cP < 0.001 versus control; dP < 0.05, eP < 0.01, and fP < 0.001 versus Aβ25–35; gP < 0.05, hP < 0.01, and iP < 0.001 versus juice A; jP < 0.05, kP < 0.01, and lP < 0.001 versus (A + Aβ).
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Related In: Results  -  Collection


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fig6: Broccoli sprouts juices upregulate the cellular antioxidant defence capacity. 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 gene expression of (a) HO-1; (b) TRX; (c) TrxR. 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. (d) Total cell lysates were subjected to spectrophotometric assay of NQO1 activity which was expressed as a percentage compared to the control. Data are represented as mean ± SEM (n = 3), aP < 0.05, bP < 0.01, and cP < 0.001 versus control; dP < 0.05, eP < 0.01, and fP < 0.001 versus Aβ25–35; gP < 0.05, hP < 0.01, and iP < 0.001 versus juice A; jP < 0.05, kP < 0.01, and lP < 0.001 versus (A + Aβ).

Mentions: To investigate the molecular mechanisms of neuroprotection exerted by broccoli sprouts juices against Aβ25–35-induced cell death, the expression or activity of various cellular antioxidant enzymes was examined. For this purpose, we determined the mRNA levels of enzymes such as HO-1, Trx, and TrxR by qRT-PCR. Moreover, we tested the activity of the enzyme NQO1 by a specific spectrophotometric assay. As shown in Figure 6(a), HO-1 gene expression in SH-SY5Y cells treated with 25 μM Aβ25–35 for 1, 4, 7, 24, 48, and 72 h showed little fluctuation. In contrast, following exposure to 10 μL/mL of juices A or B, either alone or in the presence of 25 μM Aβ25–35, HO-1 mRNA levels in the cells were strongly increased. Relative to control group, HO-1 mRNA started to increase after 4 h and up to 7 h treatment with broccoli sprouts juices. In this time frame, HO-1 gene expression increased ~12 times (P < 0.001) with juice A and 25 times (P < 0.001) with juice B compared to control, respectively. At 24, 48, and 72 h of exposure, HO-1 mRNA levels were substantially lower than at 4 h, although an appreciable HO-1 gene induction was still present compared to control cells. All through the time points, juice B was significantly more effective in upregulating HO-1 gene expression compared to juice A, both in the absence and in the presence of Aβ25–35. Trx mRNA levels were altered as a result of exposure to Aβ25–35 only from 24 h, remaining constant up to 72 h (Figure 6(b)). In any case, the magnitude of induction was rather weak (1.2 times versus control, P < 0.05). Treatment with broccoli sprouts juices, alone or in presence of Aβ25–35, significantly increased TRX gene expression starting from 1 h of incubation with juice B and at 4 h with juice A. Compared to control, during the first 7 h of exposure, the increase of Trx mRNA levels observed with juice B was higher than that caused by juice A, which only within 24 h, and up to 72 h, matched the inductive capacity of juice B. The maximum increase in TRX gene expression was observed at 72 h of treatment with broccoli sprouts juices, either in the absence or in the presence of Aβ25–35, with levels reaching 1.7 and 2.0 times (P < 0.01) that of the control for juice A and juice B, respectively. The pattern of the kinetics of TRXR gene expression was comparable to that of the above described TRX gene (Figure 6(c)). After 7 h of incubation, and up to 72 h, the treatment with 25 μM Aβ25–35 determined a modest (1.5 times) and stable increase in the level of TrxR mRNA compared to the control group (P < 0.05 at 7 and 24 h; P < 0.01 at 48 and 72 h). Already after 1 h of exposure, 10 μL/mL of juice B alone or in the presence of Aβ25–35 induced a significant increase (1.5 times versus control, P < 0.05) of TRXR gene expression levels which reached a maximum (3.6 times versus control, P < 0.05) at 4 h and kept constant up to 72 h. The induction of TRXR gene by juice A was evidenced only after 4 h of treatment, both in the absence and in the presence of Aβ25–35 (~2.6 times versus control, P < 0.01), reaching a plateau within 24 h (~3.3 times versus control, P < 0.001). At 7 h of exposure and for longer incubation periods, the effect of juice A was comparable to that of juice B. Measurement of NQO1 activity levels in SH-SY5Y cells did not show any effect of Aβ25–35 compared to control (Figure 6(d)). Otherwise, starting from 7 h of treatment with broccoli sprouts juices, either alone or in the presence of Aβ25–35, the NQO1 activity showed a time-dependent increase, with a significantly higher effect of juice B compared to juice A. The stimulating effect of juice B on NQO1 activity was higher than that of juice A from 24 h up to 72 h of exposure (24 h: 3.1 times A and (A + Aβ) versus control, P < 0.001 and 3.9 times B and (B + Aβ) versus control, P < 0.001; 48 h: 5.2 times A and (A + Aβ) versus control, P < 0.001 and 6.8 times B and (B + Aβ) versus control, P < 0.001; 72 h: 6.7 times A and (A + Aβ) versus control, P < 0.001 e 13.9 times B and (B + Aβ) versus control, P < 0.001).


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 upregulate the cellular antioxidant defence capacity. 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 gene expression of (a) HO-1; (b) TRX; (c) TrxR. 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. (d) Total cell lysates were subjected to spectrophotometric assay of NQO1 activity which was expressed as a percentage compared to the control. Data are represented as mean ± SEM (n = 3), aP < 0.05, bP < 0.01, and cP < 0.001 versus control; dP < 0.05, eP < 0.01, and fP < 0.001 versus Aβ25–35; gP < 0.05, hP < 0.01, and iP < 0.001 versus juice A; jP < 0.05, kP < 0.01, and lP < 0.001 versus (A + Aβ).
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig6: Broccoli sprouts juices upregulate the cellular antioxidant defence capacity. 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 gene expression of (a) HO-1; (b) TRX; (c) TrxR. 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. (d) Total cell lysates were subjected to spectrophotometric assay of NQO1 activity which was expressed as a percentage compared to the control. Data are represented as mean ± SEM (n = 3), aP < 0.05, bP < 0.01, and cP < 0.001 versus control; dP < 0.05, eP < 0.01, and fP < 0.001 versus Aβ25–35; gP < 0.05, hP < 0.01, and iP < 0.001 versus juice A; jP < 0.05, kP < 0.01, and lP < 0.001 versus (A + Aβ).
Mentions: To investigate the molecular mechanisms of neuroprotection exerted by broccoli sprouts juices against Aβ25–35-induced cell death, the expression or activity of various cellular antioxidant enzymes was examined. For this purpose, we determined the mRNA levels of enzymes such as HO-1, Trx, and TrxR by qRT-PCR. Moreover, we tested the activity of the enzyme NQO1 by a specific spectrophotometric assay. As shown in Figure 6(a), HO-1 gene expression in SH-SY5Y cells treated with 25 μM Aβ25–35 for 1, 4, 7, 24, 48, and 72 h showed little fluctuation. In contrast, following exposure to 10 μL/mL of juices A or B, either alone or in the presence of 25 μM Aβ25–35, HO-1 mRNA levels in the cells were strongly increased. Relative to control group, HO-1 mRNA started to increase after 4 h and up to 7 h treatment with broccoli sprouts juices. In this time frame, HO-1 gene expression increased ~12 times (P < 0.001) with juice A and 25 times (P < 0.001) with juice B compared to control, respectively. At 24, 48, and 72 h of exposure, HO-1 mRNA levels were substantially lower than at 4 h, although an appreciable HO-1 gene induction was still present compared to control cells. All through the time points, juice B was significantly more effective in upregulating HO-1 gene expression compared to juice A, both in the absence and in the presence of Aβ25–35. Trx mRNA levels were altered as a result of exposure to Aβ25–35 only from 24 h, remaining constant up to 72 h (Figure 6(b)). In any case, the magnitude of induction was rather weak (1.2 times versus control, P < 0.05). Treatment with broccoli sprouts juices, alone or in presence of Aβ25–35, significantly increased TRX gene expression starting from 1 h of incubation with juice B and at 4 h with juice A. Compared to control, during the first 7 h of exposure, the increase of Trx mRNA levels observed with juice B was higher than that caused by juice A, which only within 24 h, and up to 72 h, matched the inductive capacity of juice B. The maximum increase in TRX gene expression was observed at 72 h of treatment with broccoli sprouts juices, either in the absence or in the presence of Aβ25–35, with levels reaching 1.7 and 2.0 times (P < 0.01) that of the control for juice A and juice B, respectively. The pattern of the kinetics of TRXR gene expression was comparable to that of the above described TRX gene (Figure 6(c)). After 7 h of incubation, and up to 72 h, the treatment with 25 μM Aβ25–35 determined a modest (1.5 times) and stable increase in the level of TrxR mRNA compared to the control group (P < 0.05 at 7 and 24 h; P < 0.01 at 48 and 72 h). Already after 1 h of exposure, 10 μL/mL of juice B alone or in the presence of Aβ25–35 induced a significant increase (1.5 times versus control, P < 0.05) of TRXR gene expression levels which reached a maximum (3.6 times versus control, P < 0.05) at 4 h and kept constant up to 72 h. The induction of TRXR gene by juice A was evidenced only after 4 h of treatment, both in the absence and in the presence of Aβ25–35 (~2.6 times versus control, P < 0.01), reaching a plateau within 24 h (~3.3 times versus control, P < 0.001). At 7 h of exposure and for longer incubation periods, the effect of juice A was comparable to that of juice B. Measurement of NQO1 activity levels in SH-SY5Y cells did not show any effect of Aβ25–35 compared to control (Figure 6(d)). Otherwise, starting from 7 h of treatment with broccoli sprouts juices, either alone or in the presence of Aβ25–35, the NQO1 activity showed a time-dependent increase, with a significantly higher effect of juice B compared to juice A. The stimulating effect of juice B on NQO1 activity was higher than that of juice A from 24 h up to 72 h of exposure (24 h: 3.1 times A and (A + Aβ) versus control, P < 0.001 and 3.9 times B and (B + Aβ) versus control, P < 0.001; 48 h: 5.2 times A and (A + Aβ) versus control, P < 0.001 and 6.8 times B and (B + Aβ) versus control, P < 0.001; 72 h: 6.7 times A and (A + Aβ) versus control, P < 0.001 e 13.9 times B and (B + Aβ) versus control, P < 0.001).

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