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Cited by 165Garlic and Onion StudiesGenomics, phytochemicals, and oxidative stressSulfur Compounds in Biology

The DOI matches its Crossref record; 63 of the 65 references it cites were found in a registry or an index.

The Effects of Allicin, a Reactive Sulfur Species from Garlic, on a Selection of Mammalian Cell Lines

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Gruhlke, Martin, Nicco, Carole, Batteux, Frederic, Slusarenko, Alan·2016·Antioxidants
DOI 10.3390/antiox6010001PMC PMC5384165
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Abstract

Garlic (Allium sativum L.) has been used as a spice and medicinal plant since ancient times. Garlic produces the thiol-reactive defence substance, allicin, upon wounding. The effects of allicin on human lung epithelium carcinoma (A549), mouse fibroblast (3T3), human umbilical vein endothelial cell (HUVEC), human colon carcinoma (HT29) and human breast cancer (MCF7) cell lines were tested. To estimate toxic effects of allicin, we used a standard MTT-test (methylthiazoltetrazolium) for cell viability and 3H-thymidine incorporation for cell proliferation. The glutathione pool was measured using monobromobimane and the formation of reactive species was identified using 2′,7′-dichlorofluoresceine-diacetate. The YO-PRO-1 iodide staining procedure was used to estimate apoptosis. Allicin reduced cell viability and cell proliferation in a concentration dependent manner. In the bimane test, it was observed that cells treated with allicin showed reduced fluorescence, suggesting glutathione oxidation. The cell lines tested differed in sensitivity to allicin in regard to viability, cell proliferation and glutathione oxidation. The 3T3 and MCF-7 cells showed a higher proportion of apoptosis compared to the other cell types. These data show that mammalian cell lines differ in their sensitivity and responses to allicin.

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Figures

Scheme 1 Biosynthesis of allicin. The nonproteinogenic amino acid alliin is converted by the activity of the alliinase enzyme to allyl sulfenic acid and dehydroalanine. Two molecules of allyl sulfenic acid condense spontaneously to one molecule of allicin.
Scheme 1 Biosynthesis of allicin. The nonproteinogenic amino acid alliin is converted by the activity of the alliinase enzyme to allyl sulfenic acid and dehydroalanine. Two molecules of allyl sulfenic acid condense spontaneously to one molecule of allicin.CC BY · PubMed Central
Figure 1 Effect of garlic juice normalized to its allicin concentration (A); and synthetic allicin (B) on the viability of different cell lines in the MTT (methylthiazoltetrazolium)-assay. All data points are means of three replicates; error bars show standard deviation.
Figure 1 Effect of garlic juice normalized to its allicin concentration (A); and synthetic allicin (B) on the viability of different cell lines in the MTT (methylthiazoltetrazolium)-assay. All data points are means of three replicates; error bars show standard deviation.CC BY · PubMed Central
Figure 2 Effect of garlic juice normalized to its allicin concentration (A); and synthetic allicin (B) on cell proliferation as tested by incorporation of 3H labelled thymidine in DNA and measured by scintillation. All data points are means of three replicates; error bars show standard deviation.
Figure 2 Effect of garlic juice normalized to its allicin concentration (A); and synthetic allicin (B) on cell proliferation as tested by incorporation of 3H labelled thymidine in DNA and measured by scintillation. All data points are means of three replicates; error bars show standard deviation.CC BY · PubMed Central
Figure 3 Measurement of reduced glutathione with monobromobimane. Different cell lines were pipetted into 96-well plates (3T3 (mouse fibroblast) and HUVEC (human umbilical vein endothelial cell) 1 × 104 cells per well, A549 (human lung epithelium carcinoma), MCF7 (human breast cancer), HT29 (human colon carcinoma) 2 × 104) and treated with allicin in garlic juice (A) or chemically synthesized allicin (B) for 24 h. Monobromobimane was added to a final concentration of 50 µM and incubated for 15 min before measurement. All data points are means of three replicates; error bars represent standard deviation.
Figure 3 Measurement of reduced glutathione with monobromobimane. Different cell lines were pipetted into 96-well plates (3T3 (mouse fibroblast) and HUVEC (human umbilical vein endothelial cell) 1 × 104 cells per well, A549 (human lung epithelium carcinoma), MCF7 (human breast cancer), HT29 (human colon carcinoma) 2 × 104) and treated with allicin in garlic juice (A) or chemically synthesized allicin (B) for 24 h. Monobromobimane was added to a final concentration of 50 µM and incubated for 15 min before measurement. All data points are means of three replicates; error bars represent standard deviation.CC BY · PubMed Central
Figure 4 Detection of Reactive Species by 2′,7′-Dichlorofluoresceine-Diacetate (DCFDA) upon treatment with garlic juice (A); or synthetic allicin (B). Cells were incubated for 24 h with allicin, washed and treated with DCFDA in DMSO (dimethyl sulfoxide) to a final concentration of 50 µM for 15 min at room temperature, before fluorescence was measured at 530 nm. All data points are means of three replicates; error bars represent standard deviation.
Figure 4 Detection of Reactive Species by 2′,7′-Dichlorofluoresceine-Diacetate (DCFDA) upon treatment with garlic juice (A); or synthetic allicin (B). Cells were incubated for 24 h with allicin, washed and treated with DCFDA in DMSO (dimethyl sulfoxide) to a final concentration of 50 µM for 15 min at room temperature, before fluorescence was measured at 530 nm. All data points are means of three replicates; error bars represent standard deviation.CC BY · PubMed Central
Figure 5 Induction of apoptosis by garlic juice normalized to its allicin concentration (A); and synthetic allicin (B). Different cell lines were seeded into 96-well plates (3T3 and HUVEC 1 × 104 cells per well, A549, MCF7, HT29 2 × 104) and treated with allicin in garlic juice or chemically synthesized for 1 h. After washing the cells, YO-PRO1 iodide was added to a final concentration of 1 µM and incubated for 20 min at room temperature. Relative fluorescence was measured at 509 nm after excitation at 490 nm. All data points are means of three replicates; error bars represent standard deviation.
Figure 5 Induction of apoptosis by garlic juice normalized to its allicin concentration (A); and synthetic allicin (B). Different cell lines were seeded into 96-well plates (3T3 and HUVEC 1 × 104 cells per well, A549, MCF7, HT29 2 × 104) and treated with allicin in garlic juice or chemically synthesized for 1 h. After washing the cells, YO-PRO1 iodide was added to a final concentration of 1 µM and incubated for 20 min at room temperature. Relative fluorescence was measured at 509 nm after excitation at 490 nm. All data points are means of three replicates; error bars represent standard deviation.CC BY · PubMed Central

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Metadata

Authors
Gruhlke, M., Nicco, C., Batteux, F., & Slusarenko, A.
Affiliations
  • RWTH Aachen University (DE)
  • Hôpital Cochin (FR)
  • Laboratoire d’Immunologie biologique, Hôpital Cochin, Paris 75679, France
Corresponding author
Gruhlke, Martin
ORCID
Venue
Antioxidants
Publisher
MDPI AG
Published
2016-12-26
Volume
6
Issue
1
Pages
1
ISSN
2076-3921 (ISSN-L)
DOI
10.3390/antiox6010001
PMC
PMC5384165
OpenAlex
W2562413929
Cited by
165
References
65
Citations by year
2026: 8 · 2025: 13 · 2024: 18 · 2023: 12 · 2022: 16 · 2021: 32 · 2020: 21 · 2019: 21 · 2018: 14 · 2017: 10
FWCI
17.57
Type
journal article
Language
en
Open access
Licence
Locations
  • Antioxidants (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
  • RWTH Publications (RWTH Aachen) (submittedVersion, other-oa)
  • RWTH Publications (RWTH Aachen)
Funders
  • European Commission (215009)
  • RWTH Aachen University
Topics
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Garlic and Onion Studies
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics, phytochemicals, and oxidative stress
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Sulfur Compounds in Biology
Keywords
allicin, garlic, reactive sulfur species, cell viability, cell proliferation, glutathione oxidation, apoptosis, A549 cells, 3T3 fibroblasts, human umbilical vein endothelial cells, HT-29 cells, MCF-7 cells, cytotoxicity, cell line sensitivity

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