Elsevier

Phytochemistry

Volume 190, October 2021, 112861
Phytochemistry

Determination of phenolic profiles of Herniaria polygama and Herniaria incana fractions and their in vitro antioxidant and anti-inflammatory effects

https://doi.org/10.1016/j.phytochem.2021.112861Get rights and content

Highlights

  • 72 metabolites found in Herniaria polygama phenolic fraction and 63 in H. incana.

  • One undescribed rhamnocitrin glycoside isolated from H. incana.

  • Anti-(oxidant/inflammatory) actions of the Herniaria phenolic fractions determined.

  • No cytotoxicity of the examined fractions towards PBMCs was observed.

Abstract

The study is based on phytochemical profiling and in vitro evaluation of biological effects of phenolic acid derivatives-rich Herniaria fractions, isolated from two rupturewort (Herniaria L.) species, i.e. Herniaria incana Lam. (syn. H. besseri Fisch. ex Hornem) and H. polygama J. Gay (syn. H. odorata). For the first time, the composition of phenolic compounds of these species was extensively evaluated by both LC-HR-QTOF-ESI-MS and Nuclear Magnetic Resonance spectroscopy (NMR). LC-MS analyses of H. polygama revealed 72 tentatively identified compounds, while H. incana – 63. Only 8% of the metabolites reported in this work have been previously described for Herniaria spp. Most of the identified specialized metabolites were cinnamic and benzoic acid derivatives. Phenolic fraction of H. incana herb contained flavonoids as well. A multi-step chromatographic separation of phenolic fractions from H. polygama yielded three known cinnamic and one benzoic acid derivates, and from H. incana – 4 known flavonoids and one previously undescribed, i.e. rhamnocitrin-3-O-[3-hydroxy-3-methylglutaryl-(1 → 6′′)]-[α-rhamnopyranosyl-(1 → 2′′)]-β-glucopyranoside.

Antioxidant properties of the examined fractions (1–50 μg/ml) were assessed in human blood plasma under the conditions of peroxynitrite-induced oxidative stress. Measurements of well-known biomarkers such as 3-nitrotyrosine, protein thiol groups, thiobarbituric acid-reactive substances and the ferric reducing ability of blood plasma revealed the protective effect of Herniaria fractions against oxidative damage to blood plasma components. Furthermore, the examined fractions effectively ameliorated the inflammatory response of the concanavalin A-stimulated human peripheral blood mononuclear cells (PBMCs). Additionally, cellular safety of the fractions was confirmed in PBMCs.

Introduction

The Herniaria L. genus (rupturewort) includes well-known traditional medicinal plants used in different regions of Europe, Asia, and Africa. The ethnomedicinal significance of some species (e.g. H. glabra L.) has been confirmed by their presence in pharmacopoeias of European countries, e.g. Poland, Czech Republic, Austria, and Hungary. Ruptureworts are mainly recommended to alleviate disorders related to the urinary system, including kidney and bladder stones as well as bladder infections (Al-Snafi, 2018; Kozachok et al, 2016, 2018, 2020). Hence, the biological activity of these plants has been mainly investigated in the context of their usefulness in the treatment of nephrolithiasis or other disorders of the urinary tract. Moreover, the vast majority of available in vitro and in vivo evidence of biological activity of herbal preparations and extracts based on Heniaria is related to H. glabra L. (HG) and H. hirsuta L. (HH) (Atmani et al, 2003, 2006; Wojnicz et al., 2012), whereas biological properties, pharmacological potential, and phytoconstituents of other ruptureworts still remain poorly described. Nevertheless, ethnopharmacological guidelines indicate interchangeability of HG with other Herniaria species (Lebeda et al., 1992).

Our earlier work provided some encouraging information on the antioxidant effect of γ-pyrone compounds isolated from HG (Kozachok et al., 2018). That study, focused on flavonoids and maltol glycosides from HG, was the first report on the antioxidant activity of Herniaria-derived substances in the biological experimental system, i.e. human blood plasma in vitro. However, to our best knowledge, there is no data on either antioxidant or anti-inflammatory effects of Herniaria incana Lam. (HI, syn. H. besseri Fisch. ex Hornem.) (“Herniaria incana | International Plant Names Index,” n. d., “Herniaria incana Lam. | Plants of the World Online | Kew Science,” n. d.) and H. polygama J. Gay (HP, syn. H. odorata Andrz. Ex J. Gay) (“Herniaria polygama | International Plant Names Index,” n. d., “Herniaria polygama J. Gay | Plants of the World Online | Kew Science,” n. d.) in biological systems.

Antioxidant and anti-inflammatory properties of plant-derived compounds and mixtures significantly contribute to their beneficial effects on the human organism. Therefore, they may play an important role in the treatment and prevention of different disorders, including urolithiasis (Zeng et al., 2019). Above properties, however, remain unexplored for most of rupturewort species. Our present study covers aspects related to the aforementioned gaps in the current knowledge of phytochemical characteristics (with special emphasis on phenolic acids and their derivatives) and the determination of biological activity of Herniaria species. The available literature indicates a remarkable potential of phenolic acid derivatives (PADs) in the development of new antioxidant and anti-inflammatory agents (Lee et al., 2006; Ribeiro et al., 2019; Taofiq et al., 2015). The comparison of UHPLC-PDA chromatographic profiles of HG, HP, and HI herb (HIh) methanolic extracts showed some considerable differences in their phenolic metabolites (Fig. S32). Besides, HG extract contained a small number of PADs. Hitherto, only scant information was published about the chemical composition of HI regarding the content of essential oil (Lazari et al., 2000), and rutin (Akulshina and Darmohray, 2008). The papers by Akulshina and Darmohray (2010), 2008; Boguslavskaya et al. (1985), and Kozachok et al. (2016) focused on coumarins, flavonols and monosaccharides of HP. Furthermore, no information on PADs in H. incana Lam. and H. polygama J. Gay is available.

Our study aimed to obtain PADs-rich fractions from HP and HIh, to determine their chemical profiles and quantify the content of the individual constituents using modern chromatographic techniques such as UHPLC-DAD-QTOF-ESI-HRMS and NMR. Phytochemical profiling was complemented with biological studies on the antioxidant and anti-inflammatory potential of these fractions in the experimental system of human blood plasma exposed to the peroxynitrite-induced oxidative stress and inflammatory response of human peripheral blood mononuclear cells (PBMCs), respectively. The cellular safety of the examined Herniaria-derived fractions was confirmed in cytotoxicity assays.

Section snippets

Results and discussion

Methanolic extracts from HIh and HP were consecutively defatted with n-hexane, subjected to reversed-phase-solid phase extraction (RP-SPE) and gel permeation column chromatography. This procedure allowed obtaining PADs-rich fractions. As a result, PADs content in fractions received from HIh and HP increased 4.3 and 5.0 times, respectively, compared to their RP-SPE purified extracts. Due to insufficient information on phenolic metabolites of HI and HP, we isolated the most abundant and

Conclusions

Overall, this work provides new data on the phytochemical profile and biological activity of phenolic fractions isolated from H. polygama and H. incana. This is the first study of their activity in biological experimental systems. It was also the first time that the detailed characterization of phenolic components was performed, accompanied by NMR structural analysis. It is worth noting that only 8% of the metabolites reported here had been described previously in Herniaria spp. Additionally,

Reagents

Acetonitrile (HPLC and LC-MS grade), methanol (HPLC grade), formic acid (98–100% purity and MS-grade), deuterated MeOH-d4 (99.96% D) were purchased from Merck (Darmstadt, Germany). Peroxynitrite was synthesized according to the method described by Pryor et al., 1995. Ferulic acid (Aldrich, 99%), 5-CQA (Aldrich, ≥95%), 4-CQA (Sigma, ≥95%), and herniarin (Roth, 98%), rutin and narcissin were previously isolated from H. glabra L. (Kozachok et al., 2018). Trolox®, thiobarbituric acid, tris,

Author contributions

J. Kolodziejczyk-Czepas and S. Kozachok contributed equally to this work.

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.

Acknowledgements

This work was partly supported by grants from the National Science Centre (DEC-2017/01/X/NZ1/00086). Studies not included in the aforementioned project were financially supported by the University of Łódź, Poland (grant no. 506/1136).

The authors would like to thank Dr. Mariusz Kowalczyk (Dept. Biochemistry and Crop Quality, Institute of Soil Science and Plant Cultivation, Puławy, Poland) for developing LC-QTOF-MS analysis methods and carrying out LC-MS data acquisition, Dr. Olena Bondarenko

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