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. 2024 Jan:112:105947.
doi: 10.1016/j.jff.2023.105947. Epub 2023 Dec 15.

Chokeberry reduces inflammation in human preadipocytes

Dale C Brunelle  1 , Kate J Larson  1 , Amy Bundy  1 , James N Roemmich  1 , Donald Warne  2 , Nicole Redvers  2   3
Affiliations

Chokeberry reduces inflammation in human preadipocytes

Dale C Brunelle et al. J Funct Foods. 2024 Jan.

Abstract

Chokeberry, Aronia melanocarpa, is an indigenous fruit from North America used as food and to prevent chronic disease by Indigenous Peoples. The objective of this study was to test anti-inflammatory effects of anthocyanin on palmitic acid (PA)-induced IL-6 gene expression, IL-6 DNA methylation, and histone (H3) acetylation. Additionally, we examined effects of anthocyanins Cyanidin-3-O-galactoside (C3Gal) and Cyanidin-3-glucoside (C3G) on IL-6 gene expression. Human primary pre-adipocytes were treated with chokeberry juice extract (CBE), C3Gal or C3G in the presence or absence of PA or lipopolysaccharide (LPS). CBE inhibited LPS- and PA-induced IL-6 mRNA expression (p < 0.0001), while C3G and C3Gal had smaller effects. Human IL-6 promoter DNA methylation was increased (p = 0.0256) in CBE treated cells compared to control. Histone H3 acetylations were not affected by CBE or PA treatment. These data indicate that CBE epigenetically reduced PA-induced inflammation by regulating IL-6 DNA methylation without affecting histone modifications in human preadipocyte cells.

Keywords: Anthocyanin; Aronia melanocarpa; DNA methylation; Human preadipocyte; IL-6 promoter; Inflammation.

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Conflict of interest statement

Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Fig. 1.
Fig. 1.
Effects of treatment of CBE on inflammation induced by PA as determined by IL-6 expression from AllPrep extraction. RT-qPCR validation results for IL-6 expression in human primary preadipocytes presented as fold-change ± SEM. Fold-change is relative to the control. (A) PA and CBE treatment results. Significant (P < 0.05) effects identified through a one-way ANOVA and Tukey’s Post Hoc. RNA extracted from same source as Figs. 3 and 4.
Fig. 2.
Fig. 2.. Effects of treatments of C3G, C3Gal, and CBE on inflammation induced by PA or LPS as determined by IL-6 expression
RT-qPCR validation results for IL-6 expression in human primary preadipocytes presented as fold-change ± SEM. Fold-change is relative to the control. (A) LPS and C3G treatments results (B) LPS and CBE treatment results (C) PA and C3Gal treatment results (D) PA and CBE treatment results. Significant (P < 0.05) effects identified through a one-way ANOVA and Tukey’s Post Hoc.
Fig. 3.
Fig. 3.
Effects of treatment of CBE and PA on methylation of DNA in the IL-6 Promoter region from AllPrep extraction. Methylation analysis of four CpG islands from pyrosequenced DNA of IL-6 promoter region. (A) Position 1 (CpG −227); (B) Position 2 (CpG −212); (C) Position 3 (CpG −162); (D) Position 4 (CpG −118). Significant (P < 0.05) effects identified through a one-way ANOVA and Tukey’s Post Hoc. DNA extracted from same source as Figs. 1 and 4.
Fig. 4.
Fig. 4.
Effects of treatment of CBE and PA on Histone H3 acetylation from AllPrep extraction. Histone H3 acetylation of Lysine residues 9, 14, and 18 as determine by protein expression relative to the H3 Histone. (A) Acetylation of H3K9 (B) Acetylation of H3K14 (C) Acetylation of H3K18. Significant (P < 0.05) effects identified through a one-way ANOVA and Tukey’s Post Hoc. Protein extracted from same source as Figs. 1 and 3.
Fig. 5.
Fig. 5.. Chokeberry Extract and PA interaction
Chokeberry Extract treatment induces transcription of the Nrf2 pathway, hinders NF-kB pathway, and increases methylation of CpG Position 4. Methylation of Position 4 inhibits SP1 binding. PA treatment induces transcription of the IL-6 gene by the NF-kB pathway, but in the presence of CBE IL-6 expression is reduced.
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