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 Table of Contents  
ORIGINAL ARTICLE
Year : 2020  |  Volume : 4  |  Issue : 2  |  Page : 85-96

Effect of thymoquinone and allicin on some antioxidant parameters in cancer prostate (PC3) and colon cancer (Caco2) cell lines


1 Biochemistry Division, Department of Chemistry, Faculty of Science, Helwan University, Egypt
2 Department of Botany and Microbiology, Faculty of Science, Al Azhar University, Cairo, Egypt
3 ian Company for Blood Transfusion Services, Holding Company for Biological Products and Vaccines VACSERA, Giza, Egypt

Date of Submission20-Jul-2019
Date of Decision29-Feb-2020
Date of Acceptance01-Mar-2020
Date of Web Publication29-Jun-2020

Correspondence Address:
MSc Biochemistry Waleed A Mohamed
Faculty of Science, Ain Shams University, Egyptian Company for Blood Transfusion Services, Holding Company for Biological Products and Vaccines VACSERA, Giza
Egypt
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/sjamf.sjamf_69_19

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  Abstract 


Background Thymoquinone (TQ) and allicin are two natural compounds separated from the seeds of Nigella sativa (black seeds) and garlic, respectively. These two compounds are known to have antioxidant properties.
Aim The aim of this study was to investigate the antioxidant properties of both TQ and allicin in cancer prostate (PC3) and colon cancer (Caco2) cell lines using four cellular parameters, namely, reduced glutathione (GSH), lipid peroxide [malondialdehyde (MDA], catalase enzyme (CAT) and nitric oxide (NO).
Materials and methods Two types of cancer cell lines, PC3 cells and Caco2 cells, were used for the investigation of four aforementioned antioxidant parameters before and after treatment with the two natural compounds (TQ and allicin). Viability assay was done first to determine the concentrations used.
Results It was found that the concentration of reduced glutathione was highly significantly and significantly decreased in both PC3 cells and Caco2 cells, respectively, after treatment with TQ compared with control, whereas the change in the concentration of reduced glutathione was not significant after allicin treatment for both cell lines compared with control cells. Regarding the concentration of lipid peroxide (MDA), its concentration was highly significantly reduced in PC3 cells in both treatments, whereas it was significantly increased in Caco2 cells treated with TQ compared with control cells. Catalase activity was significantly reduced in PC3 cells after treatment with TQ, and highly significantly increased after treatment with allicin, whereas in Caco2 cells, catalase activity was highly significantly increased after treatment with TQ and allicin compared with control cells. NO concentration was not affected in PC3 cells after treatment with TQ, and highly significantly increased after treatment with allicin, while it was highly significantly increased in Caco2 cells in both treatments, compared to control cells.

Keywords: allicin, CaCo2, catalase, glutathione, malondialdehyde, nitric oxide, PC3, thymoquinone


How to cite this article:
Mahdy EM, Abdu SM, El Baseer MA, Mohamed WA. Effect of thymoquinone and allicin on some antioxidant parameters in cancer prostate (PC3) and colon cancer (Caco2) cell lines. Sci J Al-Azhar Med Fac Girls 2020;4:85-96

How to cite this URL:
Mahdy EM, Abdu SM, El Baseer MA, Mohamed WA. Effect of thymoquinone and allicin on some antioxidant parameters in cancer prostate (PC3) and colon cancer (Caco2) cell lines. Sci J Al-Azhar Med Fac Girls [serial online] 2020 [cited 2020 Sep 28];4:85-96. Available from: http://www.sjamf.eg.net/text.asp?2020/4/2/85/288289




  Introduction Top


For decades, scientists have explored the nature to search for substances that are suitable for treatment of human disorders. The search for active compounds from plant or animal sources usually results in the discovery of novel drugs. Nigella sativa or black seed has been extensively used as a spice and in folk medicine for treatment of many disorders such as ulcers, diabetes, asthma, and hypertension [1] and has recently proved efficacious against cancer in animals [2],[3],[4]. The various applications reported for N. sativa as a remedy make it a potential candidate for drug discovery. Thymoquinone (TQ) is the main constituent of the volatile oil from N. sativa seeds. It was found that TQ possesses strong antioxidant properties. Many previous studies have demonstrated that TQ has a considerable protective effect against oxidative damage induced by a variety of free radical-generating agents, including doxorubicin-induced cardiotoxicity [5]. It was also demonstrated that TQ has an analgesic and anti-inflammatory action [6], protective effect against chemically induced carcinogenesis, and an inhibiting action on eicosanoids’ generation and membrane lipid peroxidation (LPO) [7]. Researchers have proved the protective effects of TQ against asthma [8], hypertension [9], diabetes [10], and inflammation [11]. In addition, TQ is known to have antioxidant [12], analgesic, antipyretic [13], antischistosomal [14], antifungal [15], antibacterial [16], anticancer [17], anticonvulsant [18], hepatoprotective [19], and neuroprotective activities [20]. It was also found that TQ has beneficial protective effects against renal diseases; most, if not all, of these effects are attributed to the antioxidant activities of TQ [21],[22].

Garlic (Allium sativum) has been known throughout much of human history as a natural medicine [23]. Garlic contains a number of active compounds containing sulfur and phenol. These compounds have excellent antioxidant and antimicrobial activity. The most characteristic constituents of garlic are sulfur-containing compounds, especially alliin and allicin. Alliin is a sulfur-containing amino acid, which is the most representative sulfur compound in fresh garlic. It is converted to allicin by alliinase when garlic is crushed [24]. Allicin (diallyl thiosulfinate) is responsible for the typical smell of garlic. It is the best-known and most widely-studied compound [23]. It possesses antioxidant activity and exhibits hypolipidemic, antiplatelet, and procirculatory effects. Moreover, it demonstrates antibacterial, anticancer, and chemopreventive activities [25].

The aim of this study was to investigate the antioxidant properties of both TQ and allicin in cancer prostate (PC3) and colon cancer (Caco2) cell lines using four cellular parameters, namely, reduced glutathione (GSH), lipid peroxide (malondialdehyde, MDA), catalase enzyme (CAT), and nitric oxide (NO).


  Materials and methods Top


Phytochemicals

TQ was purchased from Santa Cruz Biotechnology (catalog # sc-215986; 10410 Finnell St., Dallas, Texas, USA). Allicin was prepared from garlic extract [26] as previously reported [27]. In brief, the peeled garlic bulbs were weighed (100 g), grounded thoroughly to obtain fine garlic juice, and homogenized in 100 ml of 0.9% cold and sterile saline solution in a blender at high speed for 15 min then filtered with muslin cloth. Aqueous extract of garlic was stored at –20° until use [26]. Proteins were separated using methanol (50 : 50, v/v), and supernatant was filtered through a 0.22 µm membrane. A 0.5 ml solution of filtrate was injected onto HPLC system equipped with a C18, Nucleosil 100 ODS (5 µm) semipreparative column with a dimensions of 150×10 mm. The column was eluted with methanol–water (50 : 50, v/v). The allicin in the effluents was monitored at 220 nm. The fractions containing allicin was collected into 50 ml falcon tubes and stored at –80°C until use. To concentrate allicin, collected fractions were pooled and equal volume of non-polar solvent ‘diethyl ether’ was added, and then the mixture was poured into a separating funnel; the funnel was shaken vigorously and left to stand for 15 min, until two distinct phases appeared. The organic phase was collected. Appropriate amount of double-distilled water was added and then the organic phase was evaporated under reduced pressure using a rotary evaporator at 33°C. The allicin solution was stored at –80°C for further analysis. The concentration of allicin was determined by analytical HPLC [27].

Cell lines

PC3 cells and Caco2 cells were purchased from cell culture department, the Egyptian Holding Company for Biological products and vaccines (VACSERA), Giza, Egypt.

Viability assay

Cytotoxicity for test compounds was determined on PC3 and Caco2 cell lines using MTT protocol. The effect of test compounds on cellular viability was evaluated using an assay based on the cleavage of the yellow dye MTT to purple formazan crystals by dehydrogenase activity in mitochondria, a conversion that occurs only in living cells [28],[29]. Two-fold dilutions of tested samples were made in RPMI medium with 2% fetal calf serum (maintenance medium). Overall, 0.1 ml of each dilution was tested in three different wells of 96-well tissue culture plate that was previously inoculated with 1×105 cells/ml (100 µl/well) and was incubated at 37°C for 24 h to develop a complete monolayer sheet, leaving three wells as control, receiving only maintenance medium. Plate was incubated at 37°C and examined. Cells were checked for any physical signs of toxicity, for example, partial or complete loss of the monolayer, rounding, shrinkage, or cell granulation. Then, 20 µl of MTT solution (5 mg/ml in PBS) (Bio Basic Canada Inc., Markham, Ontario, Canada) was added to each well, placed on a shaking table, 150 rpm for 5 min, to thoroughly mix the MTT into the media. Plate was incubated (37°C, 5% CO2) for 1–5 h to allow the MTT to be metabolized. The media was dumped off (plate was dried on paper towels to remove residue if necessary). Formazan (MTT metabolic product) was resuspended in 200 µl of DMSO, and then placed on a shaking table, 150 rpm for 5 min, to thoroughly mix the formazan into the solvent. Optical density of the cellular homogenate was measured at 570 nm and background at 620 nm.

Determination of antioxidant parameters

Cells were collected by centrifugation (1000–2000 rpm for 10 min at 4°C) after harvesting by using a rubber policeman rather than proteolytic enzymes. Cell pellet was homogenized in cold buffer (50 mmol/l potassium phosphate, pH 7.5, 2 mmol/l EDTA). Homogenized cells were centrifuged again at 4000 rpm for 15 min at 4°C. Supernatant was taken and stored on ice for assay.

Reduced glutathione, lipid peroxidase (MDA), catalase (CAT), and NO tests were performed according to manufacturers’ protocols and instructions [30],[31],[32],[33].

Statistical analysis

All experiments were performed three times. All statistical analyses were performed with the Statistical Package for the Social Sciences (SPSS) version 19 software (IBM SPSS Statistics, IBM, 1 New Orchard Road Armonk, New York, USA). Results are expressed as mean±SD. The statistical significance compared with control was determined by a two-tailed Student t-test, where P less than 0.05 was considered statistically significant and P less than 0.01 was considered statistically highly significant.


  Results Top


Viability assay

According to [Table 1] and [Table 2], IC50 values were found to be 623.63 and 10.87 µg/ml for both TQ and Allicin, respectively, in PC3 cell, whereas in Caco2 cells, they were found to be 389.60 and 20.46 µg/ml, respectively. [Figure 1] and [Figure 2] are light microscope images of control PC3 cells and Caco2 cells, and both cells at different concentrations of the test compounds.
Table 1 IC50 values for thymoquinone and allicin on PC3 cells

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Table 2 IC50 values for thymoquinone and allicin on Caco2 cells

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Figure 1 Viability assay for PC3 cells treated with thymoquinone and allicin. (a) Control PC3 cells; (b) effect of thymoquinone on PC3 cells at different concentrations (c) effect of thymoquinone on PC3 cells at 500 g/ml thymoquinone. CPE is shown as follows: (a) rounding and (b) granulation. (d) Effect of allicin on PC3 cells at different concentrations. (e) Effect of allicin on PC3 cells at 12 g/ml Allicin. CPE is shown as follows: (a) rounding and (b) shrinking.

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Figure 2 Viability assay for Caco2 cells treated with thymoquinone and allicin. (a) Control Caco2 cells; (b) effect of thymoquinone on Caco2 cells at different concentrations; (c) effect of thymoquinone on Caco2 cells at 500 g/ml thymoquinone CPE is shown as follows: (a) rounding and (b) granulation. (d) Effect of Allicin on Caco2 cells at different concentrations. (e) Effect of Allicin on Caco2 cells at 24 g/ml Allicin. CPE is shown as follows: (a) granulation, (b) shrinkig and (c) rounding.

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Antioxidant parameters

The IC50 values determined from viability test were used in the treatment of PC3 cells and Caco2 cells, and antioxidant parameters were determined in these treated cells compared with control. According to [Table 3] and [Figure 3], the concentration of reduced glutathione was highly significantly decreased in PC3 cells after treatment with TQ (623.63 µg/ml) compared with control, from 1.4208±0.12 mmol/l in control cells to 0.7772±0.17 mmol/l in TQ-treated cells, whereas in treatment with allicin (10.87 µg/ml), the decrease in glutathione concentration was not significant (1.2432±0.11 mmol/l), as compared with control. Regarding Caco2 cells, the concentration of reduced glutathione was significantly decreased from 0.4442±0.15 mmol/l in control cells to 0.0222±0.02 mmol/l after treatment with TQ (389.60 µg/ml) and increased insignificantly to 0.7548±0.14 mmol/l after treatment with allicin (20.46 µg/ml), as compared with control cells. The concentration of lipid peroxide (MDA) was also measured in both PC3 cells and Caco2 cells untreated and after treatment with TQ and allicin ([Table 3] and [Figure 4]). It was highly significantly reduced in PC3 cells in both treatments, from 40.23±2.99 mmol/l in control cells to 15.30±2.07 and 10.30±0.95 mmol/l in TQ –treated and allicin-treated PC3 cells, respectively, and significantly increased in Caco2 cells from 10.07±0.55 mmol/l in control cells to 20.37±2.25 mmol/l in TQ –treated cells and insignificantly increased to 15.33±2.07 mmol/l in allicin-treated cells. Catalase activity was significantly reduced in PC3 cells after treatment with TQ, from 22.50±1.25 U/l in control cells to 17.53±1.04 U/l in TQ-treated cells and highly significantly increased after treatment with allicin to 67.53±1.72 U/l. It was highly significantly increased in both treatments in Caco2 cells from 37.46±1.78 to 65.03±1.60 and 52.50±1.81 U/l in TQ-treated and allicin-treated cells, respectively, as compared with control Caco2 cells ([Table 3] and [Figure 5]). The concentration of NO was found to be the same when PC3 cells were treated with TQ (4.1667±0.22 µmol/l), and highly significantly increased when treated with allicin (8.3333±0.24 µmol/l). NO concentration was highly significantly increased in both treatments in Caco2 cells, from 4.1667±0.13 to 8.3333±0.22 µmol/l and 12.508±0.46 µmol/l in both TQ-treated and allicin-treated cells, respectively ([Table 3] and [Figure 6]).
Table 3 Concentration of reduced glutathione, lipid peroxide, catalase and nitric oxide in control, thymoquinone-treated and allicin treated PC3 cells and Caco2 cells

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Figure 3 Concentration (mmol/l) of reduced glutathione in thymoquinone-treated and allicin-treated PC3 cells and Caco2 cells, as compared with control cells.

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Figure 4 Concentration of lipid peroxide (malondialdehyde) (mmol/l) in thymoquinone-treated and aliicin-treated PC3 cells and Caco2 cells compared with control cells.

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Figure 5 Activity of catalase enzyme (U/l) in thymoquinone-treated and allicin-treated PC3 cells and Caco2 cells, compared to control cells.

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Figure 6 Concentration of nitric oxide (µmol/l) in thymoquinone-treated and allicin-treated PC3 cells and Caco2 cells compared to control cells.

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  Discussion Top


TQ was found to have antioxidant activity, which in turn was responsible for chemopreventive activities [2]. The antioxidant activity of TQ is demonstrated by scavenging anion and reactive oxygen species (ROS) [34]. It was also suggested that TQ acts as a pro-oxidant, and it potentially induces apoptosis in cancer cells [35], generates ROS, and downregulates prosurvival genes [36]. Thus, several TQ action mechanisms have shown that it acts as an antioxidant at lower concentrations and prooxidant at higher concentrations [37]. However, the situation in which TQ is present determines whether it works as prooxidant or antioxidant agent as it can be reduced to semiquinone (one reduction) or thymohydroquinone (two reductions). Thymohydroquinone acts as a strong antioxidant agent, whereas semiquinone was reported as having pro-oxidant activity generating ROS [38]. Some recent research studies stated that the accumulation of superoxide radicals in absence of detoxification system might contribute to the prooxidant effect of TQ [39]. TQ is converted into thymohydroquinone through a two-step one-electron reduction by various reductases in the mitochondria or in the microsome, or one-step two-electron reduction by a type of flavoenzyme located in the cytosol [40]. TQ can also form glutathionylated-dihydro TQ by interaction with GSH via a nonenzymatic reduction [41]. It was suggested that this interaction between glutathione and TQ gives a possible synergistic mechanism and other beneficial effects [12]. TQ also suppresses the enhanced level of LPO showed by reduced MDA level, which could be recognized as a strong antioxidant potential of TQ. The quinone structure of TQ has redox properties, which is associated with the antioxidant effect of TQ. Additionally, its capability to cross morphophysiological barriers leads to its easy access to subcellular compartments and facilitates the radical scavenging effect [42].

Allicin is a sulfur-containing natural compound with many different biological properties. It is responsible for the typical smell and taste of freshly cut or crushed garlic [43]. Allicin is a thiosulfinate that is produced as a result of damage of the garlic tissue by an enzymatic reaction. The precursor of allicin is the nonproteinogenic amino acid alliin (s-allyl-l-cysteine sulfoxide). Alliin is hydrolyzed by the enzyme alliinase as a first step to produce allylsulfenic acid and dehydroalanine. In a second step, two molecules of allylsulfenic acid condense spontaneously to produce one molecule of allicin [44].

In general, healthy cells have a negative cytosolic redox potential. The cellular redox potential is controlled largely by the GSH/GSSG redox couple (glutathione pool), along with the NAD(P)H/NAD(P)+ couples and thioredoxins. In cells, GSSG is reduced to GSH by glutathione reductase (GR) and NADPH [45]. GR is a rate-limiting enzyme in cellular redox homoeostasis [46]. Allicin is a reactive sulfur species (RSS) [47] with oxidizing properties, and it is able to oxidize thiol groups, both of protein cysteine-residues and low-molecular weight thiols like glutathione. A more oxidized glutathione pool leads to a higher cellular redox potential. Allicin can easily penetrate the cell membrane and conjugate with GSH, forming GSSA [48]. This may account for the short-term decrease in the intracellular GSH level observed following exposure to allicin (during the first 30 min of incubation) [49]. Allicin derivatives, CSSA and GSSA, up-regulated the glutathione level at a higher concentration than allicin itself. The effect of CSSA was evident after a shorter time period than GSSA, probably owing to either a higher cellular permeability or higher rate of dissociation of the former. It can be postulated that the allicin derivatives formed in the body on exposure to allicin may serve as a reservoir (buffer) for allicin-derived thiol-reactive molecules. Furthermore, the increased level of cellular glutathione represents an increase in its reduced (GSH) but not its oxidized (GSSG) form, indicating an increased antioxidant potential of the cells following exposure to allicin or its derivatives. Furthermore, cells pretreated with CSSA were found to be significantly protected from tBuOOH-induced oxidative stress and cell death [49].

Oxidation of protein thiols can lead to changes in protein structure, for example, through disulfide bond formation. Redox-triggered structural changes in proteins can lead to loss or gain of function [50]. It was found that 232 proteins in human cells could be modified upon exposure to allicin, and most of them are related to the cytoskeleton and enzymes of glycolysis, which suggests that allicin is involved in the primary metabolism of the cell directly [51]. It was also found that allicin has the ability to cross the blood-brain barrier, and it accumulates at therapeutic levels in the brain [52].

This study aims at investigating the effect of TQ and allicin on some antioxidant parameters, namely, reduced glutathione, lipid peroxide (MDA), catalase and NO in PC3 and Caco2 cell lines, as models of cancer cells.

In our study, the concentration of reduced glutathione (GSH) was highly significantly and significantly decreased in PC3 cells and Caco2 cells, respectively, when treated with TQ. These observations are consistent with many previous papers that studied the effect of TQ on reduced glutathione. For example, Rooney and Ryan [53] studied the effect of two active constituents of N. sativa against HEp-2 cancer cells. These two active constituents are alpha-hederin and TQ. In this study, it was stated that as the concentration of TQ increased, the level of GSH decreased. This effect occurred when the concentration of TQ exceeds 75 μmol/l and this is not related to apoptosis, because apoptosis occurs at concentrations lower than those that cause reduction in GSH levels. In the study by Firdaus et al. [54], the level of reduced glutathione was studied in Wistar rats stressed with arsenic and treated with TQ, among other factors, and was found to be reduced owing to arsenic stress and restored in rats pretreated with TQ. In another study, the effect of varying concentrations of glutathione on PC3 cells and C4-2B cells was studied, and in both types of cells, GSH level was decreased with increasing TQ concentration from 25 to 100 µmol/l in PC3 cells and to 150 µmol/l in C4-2B cells [55]. In our study, TQ highly significantly reduced the level of lipid peroxide (MDA) in PC3 cells, whereas the level of lipid peroxide was increased significantly in Caco2 cells treated with TQ. In a study performed by Mansour et al. [38], the level of MDA was studied – among other antioxidant enzyme systems – in mice treated with different dosed of TQ and was found to be increased in liver tissue and kidney tissue when mice were treated with 25 mg/kg TQ and decreased in both tissues when mice were treated with 50 and 100 mg/kg, whereas the level of MDA was reduced in heart tissue in all concentrations used. In another study, the effect of N. sativa L. on the level of MDA in erythrocytes of broiler chickens were studied, and it was found that it reduced the level of MDA in all concentrations used [56]. In another study, pretreatment of male NMRI rats with TQ and N. sativa oil significantly decreased LPO levels measured as MDA in hippocampus portion following cerebral ischemia-reperfusion injury (IRI) [57]. It was also demonstrated that oral administration of TQ in Wistar rats at 5 mg/kg body weight for 21 days resulted in a significant reduction of the levels of different antioxidant parameters [MPO, LPO, GSH, catalase (CAT), SOD, and NO] in collagen-induced arthritis (CIA) [58]. In a recent study, the effect of TQ on dizinone-induced hepatotoxicity in mouse model was investigated. In this study, diazinone caused significant increase in the serum levels of lipid peroxide and NO, among other parameters. Following TQ administration, a significant improvement was observed in the oxidative stress biomarkers. It was concluded in this study that the administration of TQ may prevent liver damage by preventing free radical formation in animals exposed to DZN [59].

In our study, we investigated the effect of TQ on the activity of catalase enzyme in PC3 cells and Caco2 cells. The activity of catalase enzyme decreased significantly in TQ-treated PC3 cells, whereas it was highly significantly increased in TQ-treated Caco2 cells. In a study performed by Kassab and El-Hennamy [60], which investigated the effect of TQ on the amelioration of neurotoxic effect of arsenic, the effect of TQ was studied in female rats. In this study, the activity of catalase was increased compared with control in the group of rats treated with TQ in the cerebral cortex and cerebellum and reduced in the brain stem. In our study, the effect of TQ on NO in both PC3 cells and Caco2 cells was investigated, and it was found that TQ did not affect NO in PC3 cells, as compared with control, but regarding Caco2 cells, NO concentration was highly significantly increased after treatment with TQ compared with control Caco2 cells. Al Mahmoud et al. [61] stated that TQ inhibits replication of intracellular Mycobacterium tuberculosis in macrophages and modulated NO production. In this paper, TQ was found to inhibit NO production in Mycobacterium tuberculosis which might be owing to difference between cell culture and bacteria. El-Mahomoudy et al. [62] and Bedilli et al. [63] studied the effect of TQ on NO in rat macrophages and rat model, respectively. In both papers, NO concentrations were reduced, in contrast to our findings, which might be owing to difference between in-vitro and in-vivo studies.A recent study investigated some antioxidant parameters in the whole blood or plasma of adult albino rats treated with TQ and/or tramadol. In the TQ-treated group of this study, it was found that the level of GSH was significantly increased and the level of NO was significantly decreased, which is in contrast to our study, which might be owing to the difference between the in-vitro and in-vivo systems. It was also found that the level of MDA was significantly reduced and the activity of catalase enzyme was significantly increased [64].

According to our study, allicin insignificantly affected the concentration of GSH in both PC3 cells and Caco2 cells. In a study by Limor et al. [49], allicin increased the glutathione level in a concentration and time-dependent manner up to 8 folds at a concentration of 10–20 µmol/l, after 28 h of exposure. In another study, it was stated that allicin acts as an indirect antioxidant by inducing increased levels of intracellular glutathione level via the upregulation of phase II detoxifying enzymes in a Nrf2-dependent pathway (hemeoxygenase-1, superoxide dismutase, glutathione peroxidase, glutathione-S-transferases, NAD(P)H-quinine oxidoreductase, and gamma-glutamylcysteine synthetase) [49],[65],[66]. In our study, the concentration of lipid peroxide (MDA) was significantly decreased in PC3 cells, whereas in Caco2 cells, the concentration of lipid peroxide was insignificantly increased. In a previous study, it was found that allicin induced the rise of MDA in a concentration-dependent manner [67].

In our study, allicin treatment resulted in highly significant increase of catalase activity in both PC3 cells and Caco2 cells. These results are in harmony with the results of Jayanthi et al. [68]; in this previous study, allicin was found to increase the activity of catalase enzyme in animal models. Regarding NO, its concentration was highly significantly increased in both treatments. In a previous study, it was found that allicin remakably attenuated diabetes in diabetic rats. It also induced the decrease of NO production and eNOS expression and improvement in endothelial-dependent relaxation [69].

Future research directions

According to our findings, the mechanisms of action of both TQ and allicin previously discussed, and the combination treatment with other drugs, more investigations should be continued for both compounds on other antioxidant parameters to better understand mechanisms of action of both compounds. Moreover, the effect of both TQ and allicin on enzymes, receptors, DNA and RNA involved in carcinogenesis, and in cancer cells should be investigated. Moreover, as we become aware of the toxic effects of both compounds according to our research and previous researches, researchers may begin to work on clinical trials for both compounds to be more aware of the effect of both compounds in vivo.


  Conclusion Top


According to our results, both TQ and allicin have significant effect on some of the antioxidant parameters previously discussed, indicating that both compounds are promising for treatment of a number of diseases including some types of cancer.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
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    Figures

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    Tables

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