Comparative Phytochemical, Antioxidant, and Hemostatic Studies of Extract and Four Fractions from Paulownia Clone in Vitro 112 Leaves in Human Plasma

Background: The Paulownia Clone in Vitro 112, known as oxytree or oxygen tree, is a hybrid clone of the species Paulownia elongata and Paulownia fortunei (Paulowniaceae). The oxytree is a fast-growing hybrid cultivar that can adapt to wide variations in edaphic and climate conditions. In this work, Paulownia Clone in Vitro 112 leaves were separated into an extract and four fractions (A–D) differing in chemical content in order to investigate their chemical content using LC-MS analysis. The extract and fractions were also evaluated for their anticoagulant and antioxidant properties in a human plasma in vitro. Results: The Paulownia leaf extract contained mainly phenolic compounds (e.g., verbascoside), small amounts of iridoids (e.g., aucubin or 7-hydroxytometoside) and triterpenoids (e.g., maslinic acid) were also detected. Our results indicate that the extract and fractions have different effects on oxidative stress in human plasma treated with H2O2/Fe in vitro, which could be attributed to differences in their chemical content. For example, the extract and all the fractions, at the two highest concentrations of 10 and 50 µg/mL, significantly inhibited the plasma lipid peroxidation induced by H2O2/Fe. Fractions C and D, at all tested concentrations (1–50 µg/mL) were also found to protect plasma proteins against H2O2/Fe-induced carbonylation. The positive effects of fraction C and D were dependent on the dose. Conclusions: The extract and all four fractions, but particularly fractions C and D, which are rich in phenolic compounds, are novel sources of antioxidants, with an inhibitory effect on oxidative stress in human plasma in vitro. Additionally, the antioxidant potential of fraction D may be associated with triterpenoids

The effect of iCORM-2 (100 μM, incubation time—30 min) or CORM-2 (100 μM, incubation time—30 min) and albumin (10%, incubation time—30 min) on biomarkers of oxidative stress (lipid peroxidation (A), carbonyl group formation (B), oxidation of protein thiols (C)) in plasma treated with H₂O₂/Fe.

<p>Data represents means ± SD of 5–11 experiments (from different donors).</p

The role of CORM-2 as a modulator of oxidative stress and hemostatic parameters of human plasma <i>in vitro</i>

<div><p>Purpose</p><p>The main aim of the experiment is to examine the effect of CORM-2, a donor of carbon monoxide (CO), on oxidative stress in human plasma <i>in vitro</i>. In addition, it examines the effects of CORM-2 on the hemostatic parameters of plasma: the activated partial thromboplastin time (APTT), thrombin time (TT) and prothrombin time (PT).</p><p>Methods</p><p>Human plasma was incubated for 5–60 min with different concentrations of CORM-2: 0.1–100 μM. Following this, various hemostatic factors and biomarkers of oxidative stress were studied. Lipid peroxidation was measured as thiobarbituric acid reactive substance (TBARS) concentration, and the oxidation of amino acid residues in proteins was measured by determining the amounts of carbonyl and thiol groups.</p><p>Results</p><p>Two oxidative stress inducers: hydrogen peroxide (H₂O₂) and the donor of hydroxyl radical (H₂O₂/Fe) were used. Decrease in protein carbonylation, thiol group oxidation and lipid peroxidation were detected at tested concentrations of CORM-2.</p><p>Conclusion</p><p>Our results indicate that CORM-2 may have antioxidant properties in human plasma treated with H₂O₂ or H₂O₂/Fe. In addition, our results indicate the anti-coagulant activities of CORM-2 <i>in vitro</i>.</p></div

The effect of CORM-2 (0.1–100 μM, incubation time– 30 and 60 min) on carbonyl group formation (plasma protein oxidation) induced by H₂O₂/Fe.

<p>Data represents means ± SD of 6 experiments (from different donors). * p<0.05 <i>vs</i>. control (+ H₂O₂/Fe), n.s. p>0.05 <i>vs</i>. control (+ H₂O₂/Fe).</p

Chemical structure and characteristics of CORM-2 [14; modified].

<p>Chemical structure and characteristics of CORM-2 [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0184787#pone.0184787.ref014" target="_blank">14</a>; modified].</p

The effect of CORM-2 (0.1–100 μM, incubation time– 30 and 60 min) on the oxidation of protein thiols induced by H₂O₂/Fe.

<p>Data represents means ± SD of 6 experiments (from different donors). * p<0.05 <i>vs</i>. control (+ H₂O₂/Fe), ** p<0.01 <i>vs</i>. control (+ H₂O₂/Fe), *** p<0.0002 <i>vs</i>. control (+ H₂O₂/Fe), # p<0.0005 <i>vs</i>. control (+ H₂O₂/Fe), ## p<0.00002 <i>vs</i>. control (+ H₂O₂/Fe), n.s. p>0.05 <i>vs</i>. control (+ H₂O₂/Fe).</p

The effect of CORM-2 (0.1–100 μM, incubation time– 30 min) on the hemostatic parameters of human plasma (APPT, PT and TT).

<p>Data represents means ± SD of 10 experiments (from different donors). * p<0.05 <i>vs</i>. control, n.s. p>0.05 <i>vs</i>. control.</p

The effect of CORM-2 (0.1–100 μM, incubation time: 5–60 min) on plasma lipid peroxidation induced by H₂O₂.

<p>Data represents means ± SD of 6 experiments (from different donors). * p<0.05 <i>vs</i>. control (+ H₂O₂), n.s. p>0.05 <i>vs</i>. control (+ H₂O₂); # p<0.05 <i>vs</i>. control (- H₂O₂), ## p<0.02 <i>vs</i>. control (- H₂O₂).</p

Modulation of Oxidative Stress and Hemostasis by Flavonoids from Lentil Aerial Parts

While specific metabolites of lentil (Lens culinaris L.) seeds and their biological activity have been well described, other organs of this plant have attracted little scientific attention. In recent years, green parts of lentils have been shown to contain diverse acylated flavonoids. This work presents the results of the research on the effect of the crude extract, the phenolic fraction, and seven flavonoids obtained from aerial parts of lentils on oxidative damage induced by H2O2/Fe to lipid and protein constituents of human plasma. Another goal was to determine their effect on hemostasis parameters of human plasma in vitro. Most of the purified lentil flavonoids had antioxidant and anticoagulant properties. The crude extract and the phenolic fraction of lentil aerial parts showed antioxidant activity, only at the highest tested concentration (50 μg/mL). Our results indicate that aerial parts of lentils may be recommended as a source of bioactive substances