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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 on Aβ25–35-induced apoptosis. 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 24, 48, and 72 h. (a) Morphological analysis of apoptosis was achieved by staining of cell nuclei with Hoechst 33258. Arrows indicate the apoptotic cells. Images are representative of three different experiments. (b) Quantification of the percentage of apoptotic cells by fluorescence microscopy with the DNA intercalating Hoechst 33258. Data are expressed as mean ± SEM (n = 3), aP < 0.05 and bP < 0.001 versus control; cP < 0.05, dP < 0.01, and eP < 0.001 versus Aβ25–35; fP < 0.05 versus juice A. (c) Analysis by flow cytometry of the sub-G1 peak in cells stained with propidium iodide (PI). The percentage of cells with hypodiploid DNA content is shown as function of the PI fluorescence signal (FL2-A). Histograms are representative of three independent experiments.
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fig2: Protective effect of broccoli sprouts juices on Aβ25–35-induced apoptosis. 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 24, 48, and 72 h. (a) Morphological analysis of apoptosis was achieved by staining of cell nuclei with Hoechst 33258. Arrows indicate the apoptotic cells. Images are representative of three different experiments. (b) Quantification of the percentage of apoptotic cells by fluorescence microscopy with the DNA intercalating Hoechst 33258. Data are expressed as mean ± SEM (n = 3), aP < 0.05 and bP < 0.001 versus control; cP < 0.05, dP < 0.01, and eP < 0.001 versus Aβ25–35; fP < 0.05 versus juice A. (c) Analysis by flow cytometry of the sub-G1 peak in cells stained with propidium iodide (PI). The percentage of cells with hypodiploid DNA content is shown as function of the PI fluorescence signal (FL2-A). Histograms are representative of three independent experiments.

Mentions: SH-SY5Y cells apoptosis was evaluated by fluorescence microscopy by using the nuclear dye Hoechst 33258 (Figure 2(a)). Treatment of cells with 10 μL/mL of juice A or juice B showed no proapoptotic effect compared to controls, whereas apoptosis increased from 7% in control cultures to about 18% in cells treated with 25 μM Aβ25–35 for 24 (P < 0.001), 48 (P < 0.05), and 72 h (P < 0.001). Following treatment of the cells with Aβ25–35 in the presence of 10 μL/mL of juice A or juice B, a significant reduction in apoptotic cells was observed compared to the treatment with Aβ25–35 alone (11% in the cotreatment with juice A or juice B and Aβ25–35 versus 18% in the exposure to Aβ25–35 alone at 24 (P < 0.01) and 48 h (P < 0.05); 9% in the cotreatment with juice A or juice B and Aβ25–35 versus 18% in exposure to Aβ25–35 alone at 72 h (P < 0.001) (Figure 2(b)). To confirm these data flow cytometric analyses were also performed by staining treated cells with PI (Figure 2(c)). Data obtained revealed that 25 μM Aβ25–35 treatment induced a marked increase in the sub-G1 (hypodiploid) fraction of the total cell population, compared to the control group with percentages raising from 1.2% to 6.4% at 24 h, from 0.6% to 6.3% at 48 h, and from 0.9% to 3.5% at 72 h. Flow cytometry analysis confirmed the absence of any proapoptotic effect of both juice A and juice B (data not shown). Treatment of cells with 25 μM Aβ25–35 in the presence of 10 μL/mL of juice A or juice B reduced apoptotic cells fraction from 6.4% in Aβ to 5.5% in (A + Aβ) and to 4.3% in (B + Aβ) at 24 h; from 6.3% in Aβ to 5.4% in (A + Aβ) and to 4.0% in (B + Aβ) at 48 h; finally, from 3.5% in Aβ to 2.0% in (A + Aβ) and to 2.1% in (B + Aβ) at 72 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)

Protective effect of broccoli sprouts juices on Aβ25–35-induced apoptosis. 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 24, 48, and 72 h. (a) Morphological analysis of apoptosis was achieved by staining of cell nuclei with Hoechst 33258. Arrows indicate the apoptotic cells. Images are representative of three different experiments. (b) Quantification of the percentage of apoptotic cells by fluorescence microscopy with the DNA intercalating Hoechst 33258. Data are expressed as mean ± SEM (n = 3), aP < 0.05 and bP < 0.001 versus control; cP < 0.05, dP < 0.01, and eP < 0.001 versus Aβ25–35; fP < 0.05 versus juice A. (c) Analysis by flow cytometry of the sub-G1 peak in cells stained with propidium iodide (PI). The percentage of cells with hypodiploid DNA content is shown as function of the PI fluorescence signal (FL2-A). Histograms are representative of three independent experiments.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig2: Protective effect of broccoli sprouts juices on Aβ25–35-induced apoptosis. 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 24, 48, and 72 h. (a) Morphological analysis of apoptosis was achieved by staining of cell nuclei with Hoechst 33258. Arrows indicate the apoptotic cells. Images are representative of three different experiments. (b) Quantification of the percentage of apoptotic cells by fluorescence microscopy with the DNA intercalating Hoechst 33258. Data are expressed as mean ± SEM (n = 3), aP < 0.05 and bP < 0.001 versus control; cP < 0.05, dP < 0.01, and eP < 0.001 versus Aβ25–35; fP < 0.05 versus juice A. (c) Analysis by flow cytometry of the sub-G1 peak in cells stained with propidium iodide (PI). The percentage of cells with hypodiploid DNA content is shown as function of the PI fluorescence signal (FL2-A). Histograms are representative of three independent experiments.
Mentions: SH-SY5Y cells apoptosis was evaluated by fluorescence microscopy by using the nuclear dye Hoechst 33258 (Figure 2(a)). Treatment of cells with 10 μL/mL of juice A or juice B showed no proapoptotic effect compared to controls, whereas apoptosis increased from 7% in control cultures to about 18% in cells treated with 25 μM Aβ25–35 for 24 (P < 0.001), 48 (P < 0.05), and 72 h (P < 0.001). Following treatment of the cells with Aβ25–35 in the presence of 10 μL/mL of juice A or juice B, a significant reduction in apoptotic cells was observed compared to the treatment with Aβ25–35 alone (11% in the cotreatment with juice A or juice B and Aβ25–35 versus 18% in the exposure to Aβ25–35 alone at 24 (P < 0.01) and 48 h (P < 0.05); 9% in the cotreatment with juice A or juice B and Aβ25–35 versus 18% in exposure to Aβ25–35 alone at 72 h (P < 0.001) (Figure 2(b)). To confirm these data flow cytometric analyses were also performed by staining treated cells with PI (Figure 2(c)). Data obtained revealed that 25 μM Aβ25–35 treatment induced a marked increase in the sub-G1 (hypodiploid) fraction of the total cell population, compared to the control group with percentages raising from 1.2% to 6.4% at 24 h, from 0.6% to 6.3% at 48 h, and from 0.9% to 3.5% at 72 h. Flow cytometry analysis confirmed the absence of any proapoptotic effect of both juice A and juice B (data not shown). Treatment of cells with 25 μM Aβ25–35 in the presence of 10 μL/mL of juice A or juice B reduced apoptotic cells fraction from 6.4% in Aβ to 5.5% in (A + Aβ) and to 4.3% in (B + Aβ) at 24 h; from 6.3% in Aβ to 5.4% in (A + Aβ) and to 4.0% in (B + Aβ) at 48 h; finally, from 3.5% in Aβ to 2.0% in (A + Aβ) and to 2.1% in (B + Aβ) at 72 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