Protective Effect on Human Lymphocytes of Some Flavonoids Isolated from Two Achillea Species

This study was conducted to elucidate the in vitro protective effect of five flavonoids [apigenin (1), apigenin-7- O-glucoside (2), centaureidin (3), jaceidin (4) and quercetin (5)] against chromosomal damage in mitogen-induced human lymphocytes. Using the Cytochalasin-B blocked micronucleus (CBMN) assay, in which the biomarker of chromosome breakage and/or chromosome loss is the elevated frequency of micronucleus (MN) in binucleated (BN) cells, the presence of flavonoid 2 in minimal concentration (3 μg/mL) gave a 35.5% decrease in the frequency of MN when compared with control human lymphocytes. The same concentration of flavonoids 1, 3 and 4, reduced the MN frequency by 24.4%, 28.0% and 28.0%, respectively. Higher concentrations (6 μg/mL and 10 μg/mL) seemed less effective. Flavonoid 5 (3 μg/mL) induced a slight decrease in MN frequency (5%), while higher doses (6 μg/mL and 10 μg/mL) provoked an increase of DNA damage. The comparable values for the cytokinesis-block proliferation index (CBPI) of the tested flavonoids and positive control suggested an inhibitory effect on lymphocyte proliferation. In the DPPH. scavenging assay, flavonoids 1-4 demonstrated modest activity, in a dose-dependent manner, compared with the synthetic antioxidants BHT and Trolox, while 5 exhibited comparably high antioxidative activity.

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Clastogenic Effect of Atranorin, Evernic acid, and Usnic Acid on Human Lymphocytes

Natural Product Communications ◽

10.1177/1934578x1400900419 ◽

2014 ◽

Vol 9 (4) ◽

pp. 1934578X1400900 ◽

Cited By ~ 3

Author(s):

Gordana S. Stojanović ◽

Miroslava Stanković ◽

Igor Ž. Stojanović ◽

Ivan Palić ◽

Vesna Milovanović ◽

...

Keyword(s):

Human Lymphocytes ◽

Usnic Acid ◽

Inhibitory Effect ◽

Positive Control ◽

Proliferation Index ◽

Culture Solution ◽

Antiproliferative Effects ◽

Clastogenic Effect ◽

And Control

Three lichen secondary metabolites atranorin (1), evernic acid (2), and usnic acid (3), were evaluated for their in vitro clastogenic and antiproliferative effects on human lymphocytes using the cytochalasin-B blocked micronucleus (CBMN) assay at concentrations of 2 μg/mL, 4 μg/mL and 6 μg/mL of final culture solution. The frequency of micronucleus (MN) was scored in binucleated cells, and cytokinesis-block proliferation index (CBPI) was calculated. Among the tested compounds, 3 exhibited the most prominent effect decreasing the frequency of MN in the range of 42.5% - 48.9%, that is about double of the positive control amifostin WR-2721 that reduces MN frequency for 22.0%. The effect of evernic acid was approximately equal to action of amifostin (23.2% −32.9%). Atranorin at concentrations of 2 μg/mL and 4 μg/mL decreasing the frequency of MN only for 11.1% and 1.8%, while in concentration of 6 μg/mL increases the frequency of MN for 9.6 %. The comparable CBPI values of the investigated compounds and control suggested that they did not show a statistically significant inhibitory effect on lymphocyte cell proliferation at applied concentrations.

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DNA protective activity of triterpenoids isolated from medicinal mushroom Fomitopsis betulina

Journal of the Serbian Chemical Society ◽

10.2298/jsc210401039s ◽

2021 ◽

pp. 39-39

Author(s):

Ivana Sofrenic ◽

Boban Andjelkovic ◽

Ljubodrag Vujisic ◽

Miroslav Novakovic ◽

Aleksandar Knezevic ◽

...

Keyword(s):

Protective Effect ◽

Human Lymphocytes ◽

Acid Group ◽

Positive Control ◽

Dependent Manner ◽

Protective Activity ◽

Substitution Pattern ◽

Concentration Dependent Manner ◽

Fomitopsis Betulina

Eleven 31-methylenlanostane triterpenoids, i.e. seven 21- and four 26-oic acids, as well as a lupane triterpenoid betulin, isolated from the fruiting bodies of the mushroom Fomitopsis betulina were tested for in vitro protective effect on chromosome aberrations in peripheral human lymphocytes using cytochalasin-B blocked micronucleus (CBMN) assay. Most of the tested compounds exerted a beneficial effect by reducing DNA damage of human lymphocytes more effectively than amifostine, a radioprotective agent, used as a positive control. All the tested compounds decreased MN frequency in concentration dependent manner, with the concentration of 2.0 ?g mL-1 being the most effective - with increase of the concentration the activity slightly decreases. The structure-activity relationship (SAR) studies indicated that the lanostanes containing a conjugated 7,9 (11)-diene system exhibit lower activity than D8-analogues. It was also demonstrated that the DNA protective activities within the D8-lanostane-26-oic acid group are affected by 3-substitution pattern. In the D8 series the oxygenation at C-12 or 16 as well as 21- or 26-oic acid functionality proved beneficial for in vitro protective effect on chromosomal aberrations. Betulin exhibited the lowest protective activity, but still comparable to that of amifostine.

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A search for possible cytogenetic effects of low frequency vibrations in cultured human lymphocytes

Human & Experimental Toxicology ◽

10.1177/096032719601500608 ◽

1996 ◽

Vol 15 (6) ◽

pp. 504-507 ◽

Cited By ~ 5

Author(s):

Ahmad M Khalil ◽

W. Qassem

Keyword(s):

Human Lymphocytes ◽

Low Frequency ◽

Positive Control ◽

Peripheral Blood Lymphocytes ◽

Mean Value ◽

Negative Control ◽

Proliferation Index ◽

Cell Parameters ◽

Cytogenetic Effects

1 Acute cytogenetic effects expressed as sister-chroma tid exchanges (SCE) in peripheral blood lymphocytes were investigated after in vitro exposure to low vibration regimes (5 - 30 Hz, at 0.1 cm amplitude) for 20 min intervals and compared to non-vibrated control cultures. 2 There was no significant (P<0.05) increase in the range of SCE incidence at any vibration frequency compared to the mean value of control; 6.88 ± 0.11 to 7.97±0.22 versus 6.83±0.86, respectively. 3 When cultures were incubated for 1 h in the presence of 0.5 μg ml-1 mitomycin C (MMC; the positive control) significant elevations in the SCE frequency were observed (19.81 ± 0.67), indicating the sensitivity of the system to in vitro SCE induction. 4 Other cell parameters, such as mitotic index and cell proliferation index were statistically similar in the experimental cultures and the negative control. 5 The observed differences in the investigated end points are considered within those typically found in vitro. This suggests no relation between low frequency vibrations and cytogenetic effects.

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In-VitroCarbofuran Induced Genotoxicity in Human Lymphocytes and Its Mitigation by Vitamins C and E

Disease Markers ◽

10.1155/2012/194502 ◽

2012 ◽

Vol 32 (3) ◽

pp. 153-163 ◽

Cited By ~ 15

Author(s):

Ratnesh Kumar Sharma ◽

Bechan Sharma

Keyword(s):

Dna Damage ◽

Human Peripheral Blood ◽

Human Lymphocytes ◽

Natural Antioxidants ◽

Underlying Mechanism ◽

Positive Control ◽

Peripheral Blood Lymphocytes ◽

Dependent Manner ◽

Concentration Dependent Manner

Various efforts have been made in past in order to predict the underlying mechanism of pesticide-induced toxicity usingin vitroand animal models, however, these predictions may or may not be directly correlated with humans. The present study was designed to investigate the carbofuran induced genotoxicity and its amelioration by vitamins C and E by treating human peripheral blood lymphocytes (PBLs) with different concentrations (0, 0.5, 1.25, 2.5, 3.75 and 5.0 μM) of this compound. The treatment of PBLs with carbofuran displayed significant DNA damage in concentration dependent manner. The carbofuran induced genotoxicity could be ameliorated to considerable extent by pretreatment of PBLs with equimolar (10 μM) concentration of each of the vitamins C and E; the magnitude of protection by vitamin E being higher than by vitamin C. Also, it was found that the level of protection by these vitamins was higher when PBLs were treated with lower concentrations of pesticide. The significant DNA damage as observed by H2O2, a positive control in the present study, and its amelioration by natural antioxidants (vitamins C and E) lend an evidence to suggest that carbofuran would have caused genotoxicity via pesticide induced oxidative stress.

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Jatrophane Diterpenoids With Protective Effect on Human Lymphocytes DNA

Natural Product Communications ◽

10.1177/1934578x19848168 ◽

2019 ◽

Vol 14 (5) ◽

pp. 1934578X1984816

Author(s):

Gordana Krstić ◽

Milka Jadranin ◽

Miroslava Stanković ◽

Ivana Aljančić ◽

Ljubodrag Vujisić ◽

...

Keyword(s):

Protective Effect ◽

Human Lymphocytes ◽

Structural Features ◽

Positive Control ◽

Sar Studies ◽

Structure Activity ◽

Minimal Dose ◽

Cbmn Assay ◽

Sar Study

Two sets of structurally different jatrophanes (1-11 and 13-16), jatrophane 12, and latex extract of 2 Euphorbia species (17 and 18) were tested for in vitro protective effect against chromosome aberrations in peripheral human lymphocytes using the cytokinesis-block micronucleus (CBMN) assay. Jatrophanes 1-6 in minimal doses of 1 µg/mL prominently decreased micronuclei (MN) frequency in the range 44.86% to 34.29% and manifested considerable protective effect. From the other set of jatrophanes, 13 in the same minimal dose notably decreased MN frequency by 31.05%, while extracts 17 and 18 at a concentration of 4 µg/mL remarkably decreased the frequency of MN by 37.94% and 36.12%, respectively. Jatrophanes 12, 14, and 16 showed moderate protection, while 7-11 and 15 were less active than positive control. The structure-activity relationship (SAR) studies of the tested jatrophanes (1-16) indicated the favorable position of benzoate at C-8 or C-9 (3, 4, and 13) and a preference of isobutanoyloxy group at C-3 (1-3) rather than propanoyloxy at the same position (4-6) for pronounced protective effect on human lymphocytes DNA. In a previous SAR study on 11 jatrophanes (1, 3-8, and 13-16), the same structural features in 3, 4, and 13 influenced powerful inhibition of P-gp, while growth inhibition of cancer cells was more than doubled in 1 (isobutanoyloxy group at C-3) compared to 6 (propanoyloxy at C-3).

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Effect of Hypogymnia physodes Extracts and their Depsidones on Micronucleus Distribution in Human Lymphocytes

Natural Product Communications ◽

10.1177/1934578x1300800125 ◽

2013 ◽

Vol 8 (1) ◽

pp. 1934578X1300800 ◽

Cited By ~ 2

Author(s):

Igor Ž. Stojanović ◽

Miroslava Stanković ◽

Olga Jovanović ◽

Goran Petrović ◽

Andrija Šmelcerovic ◽

...

Keyword(s):

Diethyl Ether ◽

High Performance ◽

Human Lymphocytes ◽

Positive Control ◽

Reversed Phase ◽

Proliferation Index ◽

Culture Solution ◽

Hypogymnia Physodes ◽

Photodiode Array Detection

Three lichen depsidones, physodalic acid (1), physodic acid (2), and 3-hydroxy physodic acid (3), were isolated from Hypogymnia physodes diethyl ether extract using column chromatography, and their structures determined by comparing their UV,1H and 13C NMR spectroscopic and MS data with those given in the literature, as well as with data computed by CHEM draw ultra 11 software. The contents of 1, 2 and 3 were determined in the methanol (ME), acetone (AE), and diethyl ether (EE) extracts using reversed-phase high performance liquid chromatography with photodiode array detection. The extracts, isolated depsidones 1-3 and EE fraction F23 (consisting of 90% 2 and 3, in the ratio 5.5: 1) were evaluated for their in vitro effects on chromosome aberrations in peripheral human lymphocytes using the cytochalasin-B blocked micronucleus (CBMN) assay in doses of 1 μg/mL and 2 μg/mL of final culture solution. The frequency of MN was scored in binucleated cells, and nuclear proliferation index was calculated. It was found that 1, 2, 3, F23, and EE at 1.0 μg/mL exerted a beneficial effect on lymphocyte cells giving a significant decrease of the frequency of MN in comparison with the positive control Amifostin WR-2721. Among the tested depsidones at a concentration of 1 μg/mL, 3 exhibited the most prominent effect decreasing the frequency of MN by 30.3%, followed by 2 (28.2%) and 1 (22.0%). The extracts were less effective than the isolated depsidones.

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Synthesis and Evaluation of the Antitumor Activity of Novel 1-(4-Substituted phenyl)-2-ethyl Imidazole Apoptosis Inducers In Vitro

Molecules ◽

10.3390/molecules25184293 ◽

2020 ◽

Vol 25 (18) ◽

pp. 4293

Author(s):

Zhen-Wang Li ◽

Chun-Yan Zhong ◽

Xiao-Ran Wang ◽

Shi-Nian Li ◽

Chun-Yuan Pan ◽

...

Keyword(s):

Antitumor Activity ◽

Tumor Cells ◽

Antiproliferative Activity ◽

Antitumor Agent ◽

Positive Control ◽

Selectivity Index ◽

Time Dependent ◽

Potential Candidate ◽

Dependent Manner

Novel imidazole derivatives were designed, prepared, and evaluated in vitro for antitumor activity. The majority of the tested derivatives showed improved antiproliferative activity compared to the positive control drugs 5-FU and MTX. Among them, compound 4f exhibited outstanding antiproliferative activity against three cancer cell lines and was considerably more potent than both 5-FU and MTX. In particular, the selectivity index indicated that the tolerance of normal L-02 cells to 4f was 23–46-fold higher than that of tumor cells. This selectivity was significantly higher than that exhibited by the positive control drugs. Furthermore, compound 4f induced cell apoptosis by increasing the protein expression levels of Bax and decreasing those of Bcl-2 in a time-dependent manner. Therefore, 4f could be a potential candidate for the development of a novel antitumor agent.

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In vitro study on the effect of cornin on the activity of cytochrome P450 enzymes

BMC Complementary Medicine and Therapies ◽

10.1186/s12906-021-03309-y ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Qun Zhang ◽

Zengqiang Qu ◽

Yanqing Zhou ◽

Jin Zhou ◽

Junwei Yang ◽

...

Keyword(s):

Cytochrome P450 ◽

Drug Interaction ◽

Liver Microsomes ◽

In Vitro Study ◽

Inhibitory Effect ◽

Dependent Manner ◽

Cytochrome P450 Enzymes ◽

Drug Drug Interaction ◽

Dose Dependent

Abstract Background Cornin is a commonly used herb in cardiology for its cardioprotective effect. The effect of herbs on the activity of cytochrome P450 enzymes (CYP450s) can induce adverse drug-drug interaction even treatment failure. Therefore, it is necessary to investigate the effect of cornin on the activity of CYP450s, which can provide more guidance for the clinical application of cornin. Methods Cornin (100 μM) was incubated with eight isoforms of CYP450s, including CYP1A2, 2A6, 3A4, 2C8, 2C9, 2C19, 2D6, and 2E1, in pooled human liver microsomes. The inhibition model and corresponding parameters were also investigated. Results Cornin exerted significant inhibitory effect on the activity of CYP3A4, 2C9, and 2E1 in a dose-dependent manner with the IC50 values of 9.20, 22.91, and 14.28 μM, respectively (p < 0.05). Cornin inhibited the activity of CYP3A4 non-competitively with the Ki value of 4.69 μM, while the inhibition of CYP2C9 and 2E1 by cornin was competitive with the Ki value of 11.31 and 6.54 μM, respectively. Additionally, the inhibition of CYP3A4 by cornin was found to be time-dependent with the KI/Kinact value of 6.40/0.055 min− 1·μM− 1. Conclusions The inhibitory effect of cornin on the activity of CYP3A4, 2C9, and 2E1 indicated the potential drug-drug interaction between cornin and drugs metabolized by these CYP450s, which needs further investigation and validation.

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SAT0363 DELPHINIDIN DOSE-DEPENDENTLY DIMINISHES PERIPHERAL IL-17 AND IFN-Γ PRODUCING LYMPHOCYTES IN PSORIATIC ARTHRITIS

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2020-eular.4147 ◽

2020 ◽

Vol 79 (Suppl 1) ◽

pp. 1128.1-1129

Author(s):

A. Mavropoulos ◽

S. Tsiogkas ◽

D. Skyvalidas ◽

C. Liaskos ◽

A. Roussaki-Schulze ◽

...

Keyword(s):

Psoriatic Arthritis ◽

Cell Viability ◽

Annexin V ◽

Nkt Cells ◽

Inhibitory Effect ◽

Dependent Manner ◽

Research Support ◽

Research Activities ◽

Ifn Γ

Background:Delphinidin, a dietary anthocyanidin and powerful anti-oxidant from pigmented fruits and vegetables, has broad anti-inflammatory properties. In a human skin model of psoriasis, delphinidin reduced expression of proliferative and inflammatory markers (1).Objectives:The rationale of our study was to assess whether delphinidin can in vitro suppress IL-17 and IFN-γ production in peripheral blood mononuclear cell (PBMC) subsets from patients with psoriatic arthritis (PsA).Methods:PBMCs were obtained from 24 patients with PsA attending the outpatient clinic of the Department of Rheumatology/clinical Immunology at the University General Hospital of Larissa, Greece. 16 age- and sex-matched healthy volunteers were also included in the study. Delphinidin was supplemented at a concentration ranging from 1 to 50μg/ml, one hour prior to cell stimulation. Cell viability (Annexin V staining) and innate/adaptive lymphocyte subpopulations were assessed by flow cytometry with a panel of fluorochrome-conjugated antibodies against CD56, CD3, CD4 and CD8. Intracellular expression of IL-17 and IFN-γ was measured following PMA/ionomycin stimulation for 5 hours using standard cell permeabilization protocols and monoclonal antibodies against IL-17 and IFN-γResults:Delphinidin at concentration ≥10 μg/ml sharply diminished IL-17-production by CD4(+) T cells (Th17) and CD56(+)CD3(+) (NKT) cells from patients with psoriatic arthritis and normal controls (p≤0.05). IFN-γ producing T (CD4 and CD8) cells, as well as NK and NKT cells were also dose-dependently suppressed following delphinidin pre-incubation in both patients and healthy controls. Inhibition of IFN-γ(+) cells ranged from 27 to 69% and peaked at delphinidin concentration 20-50μg/ml. The inhibitory effect of delphinidin on IL-17 and IFN-γ producing lymphocytes was not due to compromised cell viability, as assessed by annexin V binding.Conclusion:Delphinidin exerts, in a dose-dependent manner, a profound in vitro inhibitory effect on T cell and NKT cell IL-17 and IFN-γ production in PsA, and therefore, it may be used as a dietary immunosuppressant, complementary to standard treatment.References:[1]Chamcheu JC Skin Pharmacol Physiol. 2015;28(4):177-88. doi: 10.1159/000368445Disclosure of Interests:ATHANASIOS MAVROPOULOS: None declared, Sotirios Tsiogkas: None declared, Dimitrios Skyvalidas: None declared, Christos Liaskos: None declared, Aggeliki Roussaki-Schulze Grant/research support from: Received a grant to support the educational and research activities of the department from Genesis Pharma (2018), Speakers bureau: Received honoraria from Genesis Pharma and Janssen(2017) and from Roche and Pharmaserve Lilly(2018), Efterpi Zafiriou Speakers bureau: Received honoraria from Genesis Pharma, Abbvie, Novartis, Roche, Jansses(2017) and Novartis, Abbvie(2018), Dimitrios Bogdanos: None declared, Lazaros Sakkas Grant/research support from: Received a grant to support the educational and research activities of the department from Bristol-Meyers Squib, Speakers bureau: Received honoraria from Actellion(2018), Janssen(2017), Novartis(2017), Sanofi-Aventis(2018), Abbvie(2017) and Roche(2017)

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