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

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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.

No MeSH data available.


Related in: MedlinePlus

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.
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antioxidants-06-00001-f003: 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.

Mentions: Fluorescence corresponds to the quantity of reduced glutathione in the cells. The different cell lines used vary in the content of glutathione as shown in the untreated controls where it can be seen that A549, HUVEC and MCF7 cells have similar amounts of GSH and significantly less than 3T3 and HT29 cells (Figure 3A,B, p > 0.05 one-way-AnovaR, Holm–Sidak method).


The Effects of Allicin, a Reactive Sulfur Species from Garlic, on a Selection of Mammalian Cell Lines
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.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC5384165&req=5

antioxidants-06-00001-f003: 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.
Mentions: Fluorescence corresponds to the quantity of reduced glutathione in the cells. The different cell lines used vary in the content of glutathione as shown in the untreated controls where it can be seen that A549, HUVEC and MCF7 cells have similar amounts of GSH and significantly less than 3T3 and HT29 cells (Figure 3A,B, p > 0.05 one-way-AnovaR, Holm–Sidak method).

View Article: PubMed Central - PubMed

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.

No MeSH data available.


Related in: MedlinePlus