Impact of a Withania somnifera and Bacopa monnieri Formulation on SH-SY5Y Human Neuroblastoma Cells Metabolism Through NMR Metabolomic – PubMed Black Hawk Supplements

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Objectives: This study investigates the effectiveness of an herbal formulation, STRESSLESS (ST-65), which combines ashwagandha (Withania somnifera) and bacopa (Bacopa monnieri), on SH-SY5Y human neuroblastoma cells. Given the rising interest in natural compounds for neuroprotection and stress alleviation, we aimed to explore the cellular and molecular effects of this formulation. Methods: Utilizing a nuclear magnetic resonance (NMR) metabolomic approach and ultra-high-performance liquid…
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Impact of a Withania somnifera and Bacopa monnieri Formulation on SH-SY5Y Human Neuroblastoma Cells Metabolism Through NMR Metabolomic - PubMed

Impact of a Withania somnifera and Bacopa monnieri Formulation on SH-SY5Y Human Neuroblastoma Cells Metabolism Through NMR Metabolomic

Maria D’Elia et al. Nutrients. .

Abstract

Objectives: This study investigates the effectiveness of an herbal formulation, STRESSLESS (ST-65), which combines ashwagandha (Withania somnifera) and bacopa (Bacopa monnieri), on SH-SY5Y human neuroblastoma cells. Given the rising interest in natural compounds for neuroprotection and stress alleviation, we aimed to explore the cellular and molecular effects of this formulation. Methods: Utilizing a nuclear magnetic resonance (NMR) metabolomic approach and ultra-high-performance liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS), we identified key bioactive compounds in ST-65, including withanolides from ashwagandha and bacosides from bacopa. Results: Our findings indicate that ST-65 treatment significantly alters the metabolic profile of SH-SY5Y cells. Key changes included increased levels of metabolites linked to neuroprotection, energy metabolism, and antioxidant defense. Notable enhancements were observed in specific amino acids and neuroprotective compounds, suggesting activation of neuroprotective mechanisms and mitigation of stress-induced damage. Conclusions: The study reveals a complex phyto-chemical profile of ST-65 and underscores its potential as a natural active agent for addressing stress-related neurodegenerative conditions. These insights into neuronal mechanisms provide a foundation for further exploration of herbal formulations in neuroprotection.

Keywords: 1H NMR metabolomic; Bacopa monnieri; SH-SY5Y cells; UHPLC-HRMS/MS; Withania somnifera; stress.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1

UHPLC-HRMS profile of ST-65 Formulation.

Figure 2
Figure 2

Histogram graph showing the percentage of cell viability 24 h after the exposition of different concentrations of ST-65 extract (0.80–100 µg/mL). The viability variations were calculated as the percentage of viable cells in treated cultures compared to untreated ones. Results are shown as mean ± standard deviation (SD) from three independent experiments. The statistical significancy was evaluated using the one-way analysis of variance (ANOVA), followed by Bonferroni’s test, with GraphPad Prism 8.0 software (San Diego, CA, USA) assuming significance at p < 0.05. The statistical test showed the viability of cell exposed to each concentration of ST-65 is not significantly different from the control.

Figure 3
Figure 3

1D shows the NOESY spectrum of the endometabolome (A) and esometabolome (B) of SH-SY-5Y polar extracts. The spectrum was acquired, as mentioned before. Forty-one and Thirty-six metabolites were detected in the endometabolome and esometabolome, respectively.

Figure 3
Figure 3

1D shows the NOESY spectrum of the endometabolome (A) and esometabolome (B) of SH-SY-5Y polar extracts. The spectrum was acquired, as mentioned before. Forty-one and Thirty-six metabolites were detected in the endometabolome and esometabolome, respectively.

Figure 4
Figure 4

PLS-DA score plots related to the endometabolome (A) and the esometabolome (B) of treated (green) and untreated (red) cells with ST-65 formulation. VIP score graph reported the metabolites responsible for clusters endometabolome (C) and esometabolome cluster separation (D). Volcano plot analysis of metabolic changes in the cellular polar extract of endo (E) and esometabolome (F) of treated and untreated cells. Each point on the volcano plot was based on p-value and fold-change values, set at 0.05 and 2.0, respectively. Red points identify upregulated metabolites, whereas blue points identify down-regulated metabolites.

Figure 5
Figure 5

Enrichment pathways analysis. The y-axis shows the discriminating metabolic pathways in order of p-value (expressed on the x-axis as negative of logarithm) increasing. The size of the circles represents the number of hits given as an enrichment ratio.

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Grants and funding

Project funded under the National Recovery and Resilience Plan (NRRP), Mission 4 Component 2 Investment 1.4—Call for tender No. 3138 of 16 December 2021, rectified by Decree n. 3175 of 18 December 2021 of Italian Ministry of University and Research funded by the European Union—NextGenerationEU; Award Number: Project code CN_00000033, Concession Decree No. 1034 of 17 June 2022 adopted by the Italian Ministry of University and Research, CUP: D43C22001260001, Project title “National Biodiversity Future Center—NBFC”.

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Impact of a Withania somnifera and Bacopa monnieri Formulation on SH-SY5Y Human Neuroblastoma Cells Metabolism Through NMR Metabolomic – PubMed