Chemical Profiling of Polyphenolic Fraction of Cannabis sativa L. vr. Kompolti Industrial Inflorescences: Insights into Cannabidiol Neuroprotective Effects in a Cellular Model of Parkinson's Disease

Abstract

The ultra-high-performance liquid chromatography high-resolution mass spectrometry (LC-ESI-HR-MS/MS) technique was used to characterize the polyphenolic fraction of the hot water infusion (WI) of inflorescences of Cannabis sativa L. Kompolti variety, commercially used for food preparations or cosmetic purposes. On water infusion extract, we applied a multidisciplinary approach, where NMR, MS, in vitro cell-free and cell-based assays coupled with in silico studies, were used to rationalize at the molecular level the effects of the major component Cannabidiol (CBD), in a model of Parkinson's disease (PD). The phytochemical analysis by LC-MS/MS led to the tentative identification of many components belonging to different classes of polyphenols, such as phenolic acids, flavonoids, and their glycosides. CBD and cannabidiolic acid (CBDA) were also detected in good amounts in the infusion, together with several minor cannabinoids. In addition, the water infusion WI was evaluated for mineral content, total phenolic content, flavonoid content, and antioxidant capacity by DPPH and FRAP methods. Notably, our results in a cellular model of PD highlight that CBD protects against rotenone-induced cell death without recovering neuronal morphology. These biological outcomes were rationalized by an in silico approach, where we hypothesize that CBD could influence the cellular response to oxidative stress via its interaction with the Keap1/Nrf2 pathway. In summary, these results enriched the nutraceutical profile of the water infusion of the inflorescences of the Kompolti cultivar, which demonstrated a high CBD content. This study could lead to the development of dietary supplements that could help in the management of clinical symptoms related to the antioxidant activity of CBD in the pathophysiology of PD, which remains poorly characterized.

Keywords: Cannabis sativa L. vr. Kompolti; LC-MS/MS; Parkinson’s disease (PD); antioxidant effect; cannabinoids; minerals; molecular docking; nutraceuticals; phytochemicals; polyphenols.

Figure 1

LC-MS profile of n-hexane extract in positive (A) and in negative ion mode (B).

Figure 2

LC-MS profile of CHCl₃ extract in positive (A) and in negative ion mode (B).

Figure 3

LC-MS profile of n-BuOH extract in positive (A) and in negative ion mode (B).

Figure 4

LC-MS profile of H₂O extract in positive (A) and in negative ion mode (B).

Figure 5

Effects of cannabidiol extracted from Cannabis sativa L. in a cellular model of Parkinson’s disease. (A) Bright field representative images and cell count of SH-SY5Y neuronal cell line. Differentiated cells were treated with vehicle (DMSO 0.001%, Ctrl) and rotenone (Rot, 0.1 μM) without or with cannabidiol (Rot + CBD, 1 μM). After 48 h, cells were fixed, and the images were captured. The graph represents the cell number for each experimental group (expressed as a percentage of the control) by analyzing six randomly selected fields. N = 6 biological replicates. (B–E) Immunofluorescence images and semi-quantitative analysis of (B) 8-OHdG (red), (C) Nrf2 (red), (D) PPARα (red), and (E) PPARγ (green) in SH-SY5Y cells treated as previously described. N = 8–10 independent experiments. DAPI was used to counterstain cell nuclei. Data are expressed as mean ± standard deviation (SD). The dots around the SD represent the different experimental values. Statistical analysis is carried out using one-way ANOVA tests and Tukey post hoc tests. Significance is indicated as follows: * p < 0.05; ** p < 0.01; *** p < 0.001. “a” indicates statistical significance vs. Ctrl group; “b” indicates statistical significance vs. Rot group.

Figure 6

(A) Three-dimensional structure of Keap1-Nrf2 peptide (red loop) in complex with CBD. (B) Binding mode of CBD (colored by atom type: C blue, O red, polar H white). π-cation interaction is reported in green.