Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
, 24 (8)

Myrtus Polyphenols, From Antioxidants to Anti-Inflammatory Molecules: Exploring a Network Involving Cytochromes P450 and Vitamin D

Affiliations

Myrtus Polyphenols, From Antioxidants to Anti-Inflammatory Molecules: Exploring a Network Involving Cytochromes P450 and Vitamin D

Sara Cruciani et al. Molecules.

Abstract

Inflammatory response represents one of the main mechanisms of healing and tissue function restoration. On the other hand, chronic inflammation leads to excessive secretion of pro-inflammatory cytokines involved in the onset of several diseases. Oxidative stress condition may contribute in worsening inflammatory state fall, increasing reactive oxygen species (ROS) production and cytokines release. Polyphenols can counteract inflammation and oxidative stress, modulating the release of toxic molecules and interacting with physiological defenses, such as cytochromes p450 enzymes. In this paper, we aimed at evaluating the anti-inflammatory properties of different concentrations of Myrtus communis L. pulp and seeds extracts, derived from liquor industrial production, on human fibroblasts. We determined ROS production after oxidative stress induction by H2O2 treatment, and the gene expression of different proinflammatory cytokines. We also analyzed the expression of CYP3A4 and CYP27B1 genes, in order to evaluate the capability of Myrtus polyphenols to influence the metabolic regulation of other molecules, including drugs, ROS, and vitamin D. Our results showed that Myrtus extracts exert a synergic effect with vitamin D in reducing inflammation and ROS production, protecting cells from oxidative stress damages. Moreover, the extracts modulate CYPs expression, preventing chronic inflammation and suggesting their use in development of new therapeutic formulations.

Keywords: CYPs expression; antioxidants; inflammation; interleukins; nutraceuticals; oxidative stress; vitamin D.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
ROS levels after oxidative stress. The reactive oxygen species (ROS) production was evaluated in HFF1 exposed for 12 h (Panel (A)) or 24 h (Panel (B)) to ascorbic acid (AA, orange bar) or to 0.5, 0.75 and 1 mg/mL seeds (blue bars), or to 0.5, 0.75 and 1 mg/mL pulp (yellow bars) waste extracts, then induced to oxidative stress. “H2O2” (grey bar) represents HFF1 cells exposed to H2O2 alone, without a previous extracts-treatment. ROS levels of treated cells are expressed as a percentage of control untreated HFF1 (Ctrl, black bar), considered as 1. The concentrations were read as the absorbance at 529 nm emission wavelength for each sample and were expressed as mean ± SD referred to the control (* p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001).
Figure 2
Figure 2
Gene expression of proinflammatory cytokines IL-1β and TNF-α. The expression of Interleukin 1 beta (IL-1β) and Tumor necrosis factor alpha (TNF-α) was evaluated in cells pre-treated with the extracts for 12 h and 24 h and then exposed to H2O2 (Panels (A) and (B), and panels (C) and (D), respectively). HFF1 were exposed to ascorbic acid (AA, orange bar), or to 0.5, 0.75 and 1 mg/mL seeds waste extracts (blue bars) or 0.5, 0.75 and 1 mg/ml pulp waste extracts (yellows bar). H2O2 (grey bar) represents HFF1 cells exposed to only H2O2, without previous extracts treatment. The mRNA levels for each gene was expressed as fold of change (2−∆∆Ct) of mRNA levels observed in untreated HFF1 (CTRL, black bar) defined as 1 (mean ±SD; n = 6) and normalized to Glyceraldehyde-3-Phosphate-Dehidrogenase (GAPDH). Data are represented as mean ± SD referred to the control (* p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001).
Figure 3
Figure 3
Gene expression of the proinflammatory cytokines Il-8 and VEGF-A. The expression of Interleukin 8 (IL-8) and Vascular endothelial growth factor A (VEGF-A) was evaluated at 12 h and 24 h (Panels (A) and (B), and panels (C) and (D), respectively). HFF1 were exposed to ascorbic acid (AA, orange bar), or to 0.5, 0.75 and 1 mg/ml seeds waste extracts (blue bars) or 0.5, 0.75 and 1 mg/mL pulp waste extracts (yellows bar). H2O2 (grey bar) represents HFF1 cells exposed to only H2O2, without previous extracts treatment. The mRNA levels for each gene was expressed as fold of change (2−∆∆Ct) of mRNA levels observed in untreated HFF1 (CTRL, black bar) defined as 1 (mean ± SD; n = 6) and normalized to Glyceraldehyde-3-Phosphate-Dehidrogenase (GAPDH). Data are represented as mean ± SD referred to the control (* p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001).
Figure 4
Figure 4
CYP3A4 gene expression. The expression of CYP3A4 was evaluated in cells cultured for 12 h and 24 h (Panels (A,B)) in the presence of different concentrations of pulp and seeds extracts, and then exposed to H2O2. HFF1 were exposed to ascorbic acid (AA, orange bar), or to 0.5, 0.75 and 1 mg/ml seeds waste extracts (blue bars) or 0.5, 0.75 and 1 mg/mL pulp waste extracts (yellows bar). H2O2 (grey bar) represents HFF1 cells exposed to only H2O2, without previous extracts treatment. The mRNA levels for each gene was expressed as fold of change (2−∆∆Ct) of mRNA levels observed in untreated HFF1 (CTRL, black bar) defined as 1 (mean ± SD; n = 6) and normalized to Glyceraldehyde-3-Phosphate-Dehidrogenase (GAPDH). Data are represented as mean ± SD referred to the control (* p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001).
Figure 5
Figure 5
CYP27B1 gene expression. The expression of CYP27B1 was evaluated in cells cultured for 12 h and 24 h (Panels (A,B)) in the presence of different concentrations of pulp and seeds extracts, and then exposed to H2O2. HFF1 were exposed to ascorbic acid (AA, orange bar), or to 0.5, 0.75 and 1 mg/mL seeds waste extracts (blue bars) or 0.5, 0.75 and 1 mg/mL pulp waste extracts (yellows bar). H2O2 (grey bar) represents HFF1 cells exposed to only H2O2, without previous extracts treatment. The mRNA levels for each gene was expressed as fold of change (2yw) of mRNA levels observed in untreated HFF1 (CTRL, black bar) defined as 1 (mean ± SD; n = 6) and normalized to Glyceraldehyde-3-Phosphate-Dehidrogenase (GAPDH). Data are represented as mean ± SD referred to the control (* p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001).
Figure 6
Figure 6
Regulatory activity of Myrtus products. Myrtus extracts exert antioxidant and anti-inflammatory activities against oxidative stress. They reinforce, acting in a synergic manner together with the physiological balancing systems and with other natural molecules, such as vitamin D.
Figure 7
Figure 7
Hydroxyl radical scavenging activity of myrtle byproducts. Results are expressed as EC50 (n = 3).

Similar articles

See all similar articles

Cited by 2 PubMed Central articles

References

    1. Chen L., Deng H., Cui H., Fang J., Zuo Z., Deng J., Li Y., Wang X., Zhao L. Inflammatory responses and inflammation-associated diseases in organs. Oncotarget. 2018;9:7204–7218. doi: 10.18632/oncotarget.23208. - DOI - PMC - PubMed
    1. Kotas M.E., Medzhitov R. Homeostasis, Inflammation, and Disease Susceptibility. Cell. 2015;160:816–827. doi: 10.1016/j.cell.2015.02.010. - DOI - PMC - PubMed
    1. Neri M., Fineschi V., Paolo M., Pomara C., Riezzo I., Turillazzi E., Cerretani D. Cardiac Oxidative Stress and Inflammatory Cytokines Response after Myocardial Infarction. Curr. Vasc. Pharmacol. 2015;13:26–36. doi: 10.2174/15701611113119990003. - DOI - PubMed
    1. Hunter P. The inflammation theory of disease. the growing realization that chronic inflammation is crucial in many diseases opens new avenues for treatment. EMBO Rep. 2012;13:968–970. doi: 10.1038/embor.2012.142. - DOI - PMC - PubMed
    1. Wojdasiewicz P., Poniatowski Łukasz A., Szukiewicz D. The Role of Inflammatory and Anti-Inflammatory Cytokines in the Pathogenesis of Osteoarthritis. Mediat. Inflamm. 2014;2014:1–19. doi: 10.1155/2014/561459. - DOI - PMC - PubMed

MeSH terms

Feedback