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Protective role of a melon superoxide dismutase combined with gliadin (GliSODin) on the status of lipid peroxidation and antioxidant defense against azoxymethane-induced experimental colon carcinogenesis

1 Department of Biology University El hadj Lakhder-Batna, University El Hadj Lakhder-Batna, Batna, Algeria
2 Department of Biology, University of Tebessa, University Larbi Tebessi, Tebessa, Algeria
3 Department of Animal Biology University, University of Badji Mokhtar Annaba, Laboratory of Environmental Bio Surveillance, University of Badji Mokhtar-Annaba, Annaba, Algeria

Date of Submission13-Mar-2019
Date of Decision13-May-2019
Date of Acceptance17-Oct-2019
Date of Web Publication11-Jun-2020

Correspondence Address:
Kheireddine Ouali,
Laboratory of Environmental Bio Surveillance, University of Badji Mokhtar-Annaba, BP 12 El Hadjar, Annaba 23000
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/jcrt.JCRT_175_19

 > Abstract 

Background: Azoxymethane (AOM) is a potent carcinogenic agent commonly used to induce colon cancer in rats and mice, with the cytotoxicity of AOM mediated by oxidative stress.
Aim of Study: This study investigated the protective effect of a natural antioxidant (GliSODin) against AOM-induced oxidative stress and carcinogenesis in rat colon.
Methods: Twenty male Wistar rats were randomly divided into four groups (five rats/group). The control group was fed a basal diet. AOM-treated group (AOM) was fed a basal diet and received intraperitoneal injections of AOM for 2 weeks at a dose of 15 mg/kg. The GliSODin treatment group (superoxide dismutase [SOD]) received oral supplementation of GliSODin (300 mg/kg) for 3 months, and the fourth combined group received AOM and GliSODin (AOM + SOD). All animals were continuously fed ad libitum until the age of 16 weeks when all rats were sacrificed. The colon tissues were examined microscopically for pathological changes and aberrant crypt foci (ACF) development, oxidant status (lipid peroxidation-LPO), and enzyme antioxidant system (glutathione [GSH], GSH-S-transferase, catalase, and SOD).
Results: Our results showed that AOM induced ACF development and oxidative stress (GSH depletion and lipid peroxidation) in rat colonic cells. The concomitant treatment of AOM with GliSODin significantly ameliorated the cytotoxic effects of AOM.
Conclusion: The results of this study provide in vivo evidence that GliSODin reduced the AOM-induced colon cancer in rats, through their potent antioxidant activities.

Keywords: Aberrant crypt foci, azoxymethane, colon cancer, GliSODin, oxidative stress, rat

How to cite this URL:
Baba-Ahmed F, Guedri K, Trea F, Ouali K. Protective role of a melon superoxide dismutase combined with gliadin (GliSODin) on the status of lipid peroxidation and antioxidant defense against azoxymethane-induced experimental colon carcinogenesis. J Can Res Ther [Epub ahead of print] [cited 2021 Dec 5]. Available from: https://www.cancerjournal.net/preprintarticle.asp?id=286537

 > Introduction Top

Cancer of the large and small intestine is among the main cancers involved in global morbidity and mortality.[1] Although colon cancer has been well studied, the progress made in the prevention or cure of this disease is not significant. Epidemiological and experimental studies suggest that colon cancer is strongly influenced by nutrition, especially by the composition of dietary fat.[2] Recent epidemiological studies revealed that regular consumption of fruits and plant extract improves overall human health and well-being by preventing noncommunicable diseases including several types of cancer.[3],[4]

Many studies support the hypothesis that oxidative stress is involved in pathological mechanisms and processes such as cancer.[5],[6],[7],[8] Oxidative stress leads to an increase in markers of the presence of free radicals, which react with substrates that can be oxidated (glucose, proteins, and fatty acids) and generate carbonyl reagents, the cellular effects of which are multiple as follows: protein oxidation, lipid peroxidation, modification of gene expression, and DNA modification.[9],[10],[11] Persistent oxidative stress leads to cellular overproduction of reactive oxygen species (ROS) and reactive nitrogen species, causing DNA damage that is also involved in cancer development. This theme focuses on integrating these interactions in the field of oncology where many studies are interested in establishing relationships between cancer and environmental variation and explaining its mechanisms.[12] Chemopreventive agents are called upon to exert their anticarcinogenic action by modulating lipid peroxidation (LPO) and antioxidant status in the liver.[13],[14] Several reports suggest that phenolic compounds may also act as a chemopreventive adjuvant by trapping carcinogenic agents induced by oxidative stress.[15],[16] Flavonoids, which are natural polyphenolic compounds, possess a wide range of pharmacological properties, including anticarcinogenic and anti-inflammatory properties.[17],[18],[19]

Azoxymethane (AOM) is an inducer of experimental tumors in mice and rats.[20] AOM is metabolized in the liver to methylazoxymethanol (MAM), which is catalyzed by cytochrome P450E1.[21] The conjugation of MAM with the highly reactive electrophile methyl diazoniumn ion occurs in the liver, causing intense oxidative stress. This cellular nucleophile of methylate damages the DNA.[22],[23] The mutations acquired in the DNA result in the proliferation of cells, leading to cell carcinogenesis.[24] AOM has also been reported to cause oxidative stress in the mucosa of the colon.[25]

ROS are known to increase in chronic diseases of the gastrointestinal tract,[26] but the role of oxidative stress as cell inducer of colorectal cancer remains unclear. Changes in some antioxidant system parameters in colon cancer have been reported.[27] In addition, the improvement of oxidative damage by trapping free radicals formed during oxidative stress could protect cells against carcinogenicity.[28] During carcinogenesis, the following two essential points have been reported: high levels of lipid peroxidation (LPO) and depletion of enzymatic and nonenzymatic antioxidants.[29]

In the present work, we aimed to study the initiator and promoter effects of AOM in Wistar rats receiving chemoprotective treatment with an antioxidant GliSODin, which is a melon superoxide dismutase (SOD) and wheat gliadin combination. The first objective of this experimental study was to determine the degree of involvement of free radicals in the evolution of colonic carcinogenesis in this animal model whose intestinal ecology is close to that of the human colon. These effects were observed in our study from preneoplastic lesions, microadenomas.[30] The second goal was to evaluate the effect of increased enzymatic radical scavenging by treatment with GliSODin. We hypothesize that mitigation of the intensity of oxidative stress using GliSODin could have a prophylactic effect on colonic carcinogenesis in this model.

 > Methods Top


Male Wistar rats obtained from Pasteur Institute (Algiers, Algeria) aged 4 weeks were used in this experiment. The rats were housed in individual polypropylene cages and were provided with standard laboratory chow diet and normal tap water ad libitum. Rats were kept under standard conditions (i.e., temperature 22°C ± 2°C, relative humidity 60%) and a 12-h light/dark cycle.

Experimental procedure

All experimental procedures were conducted according to the International Guidelines for Laboratory Animal Care and Use (Council of European Communities JO86/609/CEE) and were approved by the University's Ethics Committee.

Twenty male Wistar rats were randomly divided into four groups (five rats/group). Control group was fed a basal diet. AOM-treated group (AOM) was fed a basal diet and received AOM intraperitoneal injections. SOD-treated group (SOD) received GliSODin by gavage for 90 days. The combination group AOM–SOD received GilSODin and AOM. The dosing is outlined below.

Body weight was recorded weekly for the whole duration of the experiment (12 weeks).

After 12 weeks, the animals were sacrificed by decapitation under diethyl ether anesthesia after an overnight fast, and the colon tissues were removed for subsequent analysis: (1) microscopic examination for aberrant crypt foci (ACF) numeration or any other morphological changes and (2) biochemical analysis of the colonic tissue homogenates included reduced glutathione (GSH), GSH-S-transferase (GST), catalase (CAT), SOD, and lipid peroxidase (LPO) measurement.


  • AOM: Rats in the AOM and AOM–SOD groups were given two intraperitoneal injections of AOM (Sigma Chemical Co., St. Louis, MI, USA) dissolved in physiological saline once a week (15 mg/kg of body weight) for 2 weeks
  • GliSODin: Rats in the SOD and AOM–SOD groups were administered oral GliSODin at 300 mg/kg/day for 90 days. The solution is prepared extemporaneously in 9% NaCl.

Colon preparation

The colons were carefully removed from the rats and were kept on a glass plate in ice jackets. The colons were then opened longitudinally, were rinsed with ice-cold physiological saline, were sectioned longitudinally into two halves of equal width, and were spread out with flat mucosal side up. The mucosal layer from one half was removed by scraping and was immediately homogenized. The other half was fixed flat in 10% buffered formalin between two filter papers for 48 h before ACF enumeration.

ACF are known to be precursor lesions for colonic tumors. Enumeration and microscopic examination of ACF is a measure of carcinogenesis, and the method used to perform this in the present study was validated by Bird.[30] Fixed colons were stained with 0.2% methylene blue in Kreb's ringer bicarbonate buffer for 20 min in a Petri dish and rinsed with physiological saline. After staining, the colons were placed (i.e., mucosal surfaces) up on a slide, examined with a light microscope under ×40, and scored for ACF. In brief, the ACF were distinguished from normal crypts by their darker stain, enlarged and slightly elongated size, thick epithelial lining, slightly elongated cryptal opening, and slit shapes. The total number of ACF was recorded for all the examined colons.

Histopathological examination

Specimens of colon were taken from rats of different groups directly after sacrifice. They were fixed in 10% neutral buffer formalin, embedded in paraffin, sectioned at 5 μ, and stained with hematoxylin and eosin stain (H and E). The stained sections were examined under a light microscope Leica Microsystems Schweiz AG, CH-9435 Heerbrugg) and photographed using a digital camera (Leica ICC50 W).

Determination of LPO

The lipoperoxides were measured according to the method described by[31] using a Bioxytech kit (LPO-586, Cayman USA) based on the estimation of malondialdehyde (MDA), an intermediate product of lipid peroxidation, using thiobarbi-turic acid and the absorbance was read at 532 nanometers. LPOs were expressed in nmole MDA released/min/mg protein.

Determination of superoxide dismutase

The enzymatic activity of SOD in colon was assessed by the method of Oberley and Spitz.[32] For that, 50 μl of the matrix fraction was added to a mixture composed of 2 ml of the reactive medium (sodium cyanide 10−2 M, solution of nitroblue tetrazolium [NBT] at 1.76 × 10−4 M, ethylenediaminetetraacetic acid 66 mmol, methionine 10−2 M, and riboflavin 2 μmol, pH 7.8). This mixture was exposed to light of a 15-W lamp for 30 min to induce the photoreaction of riboflavin. Reduction of NBT into formazan yielded a blue color. The color was measured by a spectrophotometer at 560 nm. The enzymatic activity is calculated in terms of international unit per milligrams of proteins.

Determination of catalase activity

The CAT activity was determined according to the method of Aebi.[33] The H2O2 decomposition rate was followed by monitoring absorption at 240 nm. One unit of CAT activity is defined as the amount of enzymes required to decompose 1 μmol of hydrogen peroxide in 1 min. The enzymatic activity was expressed as μmol H2O2 consumed/min/mg protein.

Determination of reduced glutathione

Colon-reduced GSH assay: The GSH content was estimated according to the method of Weckbecker and Cory.[34] Briefly, 1.0 mL of the supernatant was precipitated with 1.0 mL of 4% sulfosalicylic acid for 1 h at 4°C. The samples were then centrifuged at 1.200 g for 15 min at 4°C. 1 mL of the supernatant was then mixed with 0.2 mL of 5-5 dithio-bis-nitrobenzoic acid (0.01 M) and 2.7 mL of phosphate buffer (pH 8.0). Immediately, the absorbance of the reaction product was measured at 412 nm. The results were expressed as nmol GSH−1 mg of protein.

Determination of glutathione-S-transferase

GST catalyzes the conjugation reaction with GSH in the first step of mercapturic acid synthesis. The activity of GST was measured according to the method of Habig et al.[35] The P-nitro benzyl chloride was used as the substrate. The absorbance was measured at 340 nm at 30 s intervals for 3 min.

Carcinoembryonic antigen and cytokine determination

Quantitative determination of carcinoembryonic antigen (CEA) concentration in plasma was performed byin vitro enzyme-linked immunosorbent assay (ELISA) using quantitative CEA ELISA Abcam's kit (CEA-96, No. ab99992). Cytokines (interleukins [IL] 1, 6, and 10 and tumor necrosis factor [TNF]) were also measured by the same method, using Cayman kits (Cayman Chemical company, Ann, Arbor, MI, USA).

Statistical analysis

The results were expressed as means ± standard deviation. Analysis of variance (ANOVA), two-way ANOVA analysis, and the Tukey's test were used to compare the mean between the groups and between control groups.

 > Results Top

Variation of body weight and body of liver and rate in control and treatment groups

Our result shows a statistically significant increase in body weight in rats treated with AOM and AOM + SOD groups (AOM: 259.2 ± 46.39 vs. AOM + SOD: 238.2 ± 229.58) compared to control rats (control: 201 ± 13.89) (P < 0.05); hyperplasia of liver and spleen, respectively (AOM: 8.18 ± 1.18 vs. AOM + SOD: 7.504 ± 1.10) compared to control rats (control: 6.058 ± 0.47) (P < 0.05); and AOM and AOM + SOD lots (AOM: 0.924 ± 0.18 vs. AOM + SOD: 0.820 ± 0.17) compared to control rats (control: 0.804 ± 0.12) (P < 0.05) [Table 1].
Table 1: Variation of body, liver, and spleen weight in control and treatment groups

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Morphometric study

This study was conducted to evaluate the effect of GliSODin on the evolution of the carcinogenic process relating to the appearance of precancerous alterations following the injection of AOM. This study is based on the quantification of ACF at the colon level according to the method used by Bird (1987) [Figure 1], [Figure 2], [Figure 3].
Figure 1: Macroscopic view of azoxymethane-treated rat colons (c-f) showing tumor presence (*) compared to control colon (a) and superoxide dismutase-treated colon (b). (f) Distal part of a colon treated with azoxymethane revealing adenomas (*)

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Figure 2: Microscopic view of birds' methylene blue-stained colon of rats treated with azoxymethane. (c-f) Aberrations of aberrant crypt foci (arrow) compared to control and glisodine-treated (a and b) colonists. In c, aberrant crypt foci at 2, 3, and 4 crypts (arrow). (d) Multiple aberrant crypt foci >4 crypts. (e) early adenoma (star). (f) Multicryptic aberrant crypt foci of colon treated with azoxymethane + superoxide dismutase. ×75 and × 150

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Figure 3: Effect of GliSODin in number and multiplicity of aberrant crypt foci induced by azoxymethane in rats

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Effect of GliSODin in number and multiplicity of aberrant crypt foci induced by azoxymethane in rats

Our results reveal a significant elevation of incidence of precancerous lesions in AOM-treated group compared with control and SOD groups. In addition, the administration of GliSODin attenuated this frequency. Furthermore, the total number of ACF per colon in the AOM–SOD group statistically significantly decreased compared to the AOM group (P < 0.05). In addition, our results clearly show that GliSODin decreases the multiplicity of neoplastic lesions; this appears in the significant decrease in multiple ACF >4 crypts [Figure 3].

Histopathological changes

The histological topographies of ACF and normal colon tissue were also evaluated by H and E staining. Histological examination of the colon sections from control and SOD groups showed normal architecture with regular mucosal and submucosal layers [Figure 4]a, [Figure 4]b, [Figure 4]c, [Figure 4]d. Mucosa from the AOM group showed proliferating mucosal glands with severe dysplastic changes representing transformation to carcinoma in rats [Figure 4]e, [Figure 4]f, [Figure 4]g, [Figure 4]h. Animals from the AOM–SOD group showed mild changes of absorptive and goblet cells [Figure 4]i and [Figure 4]j, which appeared distinctly different to the mucosa from AOM animals.
Figure 4: Longitudinal sections of the control rat colon stained with H and E. (a) General view showing the presence of the chorion extending between the crypts and forming the central axis of each villus (×150). (b) More detailed view showing the different layers, the muscular mucosa, and the submucosa (×300). (c) The serous glands and (d) the serous layer with the presence of goblet cells (black arrow) (×600). (e-h) Sections of the colon in azoxymethane rats stained E: appearance of several aberrant crypts (×150). (f) Very marked hyperplasia of the two layers of the muscularis mucosa and the submucosa (black arrows) (×300). (g) Tubular hyperplasia, which is synonymous with a precancerous lesion and formation of a conjunctivo-vascular axis (*) (×300). (h) Detailed view of a villous tubular adenoma (AdTV) (×600). Sections of azoxymethane-superoxide dismutase rat colon stained with H and E. (i) General view of the colon still showing the presence of tubular hyperplasia (V) but less important than AOM rats (×150) with decreased volume of villi (large arrow) and decrease in thickness of layers (*) at the section J (×300). (j) decreased volume of villi (double arrow) and decrease in thickness of layers (asterix)

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Colon oxidative stress status

The results of the assessment of the different markers of redox status in colon and plasma cancer marker (CEA) of rats after AOM injection are summarized in [Table 1]. GSH, an important antioxidant enzyme, had a significant decrease in the AOM group compared to the control group. At the same time, the administration of GliSODin in the AOM–SOD group significantly mitigated this effect. Similarly, GST, another key antioxidant enzyme which could regulate the level of the ROS in colon, significantly decreased in the AOM group compared to control rat, but the administration of GliSODin in the AOM–SOD group reduced this impact as well.

A significant decrease was observed in colon CAT and SOD activities in the AOM group in comparison to control and SOD rats, whereas the administration of GliSODin significantly increased CAT and SOD levels. LPO values were significantly increased in AOM-treated group and decreased in AOM + SOD-treated rats compared to control rats. This biochemical response of colon–AOM tissue was associated with a significant increase of plasma CEA concentration compared to the control and SOD groups. The administration of GliSODin to rats receiving AOM (AOM–SOD group) causes a significant decrease of CEA level [Table 2].
Table 2: Change in oxidative stress parameters such as glutathione, lipid peroxidation, catalase, glutathione-S-transferase, and superoxide dismutase in colon of control and treated rats with the azoxymethane, GliSODin (superoxide dismutase), and the combination (azoxymethane-superoxide dismutase) after 90 days of treatment

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Inflammatory status

As shown in [Table 3], IL-6 significantly increased in AOM animals compared to the control and SOD groups. The treatment with GliSODin for 12 weeks at dose of 300 mg/kg was able to significantly decrease their levels compared to the AOM-induced ACF group. Regarding the IL-1, AOM animals also showed significant increase in their levels in comparison to the control group, and the treatment with vegetal SOD was able to reduce these levels. In the AOM group, IL-10 was significantly lower compared to control group, and the animals treated with GliSODin at a dose of 300 mg/kg/day had their levels increased significantly. Our results also showed significantly elevated TNF-α concentrations in AOM rats compared to the control and SOD groups.
Table 3: Change in inflammatory status parameters such as interleukin-1, interleukin-6, interleukin-10, tumor necrosis factor, and carcinoembryonic antigen in plasma of control and treated rats with the azoxymethane, GliSODin, and the combination (azoxymethane/superoxide dismutase) after 90 days of treatment

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 > Discussion Top

The availability of oxygen determines the function of complex multicellular organisms. However, oxygen metabolism also generates toxic products called ROS. ROS can cause cellular damage through the oxidation of several essential molecules such as proteins, lipids, or DNA.[36]

The release of the free radicals following environmental oxidative stress makes the colon cells even more susceptible to apoptotic lesions such as ACF and colorectal cancer, with concomitant release of free radicals, a cascade probably mediated by the gastrointestinal mucosal defensive system against the aggressive insults. The abundance of lipid peroxidation following free radical accumulation is also implicated to cell cycle disruption, colon tissue hypertrophy, and submucosal glandular dysplasia as relative to the formation of extramucosal polyps and in correspondence to the first defense line. Recent laboratory-based evidence claimed the formation of colon cancer to the underlying free radicals/lipid peroxidation system and the consequent pathological events. The cancer expansion and invasion beyond the colon intraluminal mucosa is multifactorial and claimed in part to the imbalance of the cellular homeostasis including the disturbance of MDA, SOD, and GST production.[37]

Antioxidants such as SOD, CAT, or GSH peroxidase can be synthesized invivo, and some nonenzymatic antioxidants can be ingested through diet (e.g., -carotene or a-tocopherol).

AOM induces colon cancer in experimental animals, in a mechanism that is mediated by GSH depletion and impairing total antioxidant capacity in colonic cells of rats.[38],[39]

In this line, we were interested to find out some antioxidant, antiproliferative, and anticarcinogenic remedies from natural products to combat colon cancer, and we investigated the chemical and biological properties of the GliSODin on colorectal cancer AOM-induced ACFin vivo in order to determine the possible chemoprotective effect.

Chronic induction of ROS is linked to inflammation, which can mediate other pathologies such as cancer.[40] Tumor cells display reduced SOD activity, and overexpression of this enzyme can decrease malignancy.[41] A report on a mouse model for fibrosarcoma proposes that the SOD–Gliadin complex decreases metastasis development, which is correlated with a reduction of oxidative stress in the tumor tissue.[36] In this cancer model, QR-32 tumor cells and a gelatin sponge were co-implanted to promote inflammation and tumor development in C57 BL/6 mice. In tumors from the SOD + gliadin-treated group, SOD activity was approximately doubled. However, no differences were registered for inflammatory cell infiltration at the tumor site.

In addition, although primary tumor growth was not significantly altered, metastatic potential could be inhibited in tumor cells derived from SOD + gliadin-treated animals. In 2004, results from a study claimed that the SOD + gliadin formulation has anti-inflammatory properties.[42] The assumptions were based on the observation that in murine models, encapsulated SOD supplementation induced IL-10 production. Upregulation of this anti-inflammatory cytokine also resulted in decreased production of TNF-α, therefore reducing pro-inflammatory responses.

In this study, all AOM-treated rats, with or without SOD, developed ACF after 12 weeks of treatment. However, the number of tumors decreased significantly after the oral administration of GliSODin. Regarding the formation of ACF at 12 weeks after injection AOM, chronic supplementation with SOD rats significantly attenuated the number of ACFs and the multiplicity of foci compared to control. This attenuation effect of GliSODin on ACF formation is supported by previous studies that have shown that dietary intake of antioxidants significantly decreases ACF formation.[43],[44]

These results suggest that ingestion of the melon SOD plant extract combined with gliadin attenuated carcinogenesis in rat colon primarily at the initiation phase and to a lesser extent in the promotion phase. The suppressive effect of melon SOD + gliadin on carcinogenesis could be caused by its antioxidant property or maybe by its anti-inflammatory effect. Our results demonstrate an increase in lipid peroxidase (LPO), associated with a decrease in the activity of antioxidant enzymes SOD, CAT, and GST, suggesting the establishment of a state of oxidative stress. Oral administration of GliSODin in rats treated with AOM not only attenuated the LPO but also improved the level of GSH and the activities of GST, CAT, and mainly SOD. These antioxidant enzymes belonging to the detoxification system play an important defensive role against ROS and lipid peroxidation. Specifically, these enzymes are involved in the direct elimination of reactive oxygen metabolites, responsible for the onset of various diseases including colon cancer.[45],[46]

In the present study, these antioxidant enzymatic activities were increased in rats treated with GliSODin. These results are in agreement with those reported by some studies,[47],[48],[49],[50],[51],[52] showing that antioxidants such that berberine, luteolin, GSH, hespertin, morine, and hesperidin decrease the number and the multiplicity of ACFs by enhancing the activity of antioxidant enzymes. This indicates that GliSODin may modulate the proliferative activity of crypts, thereby decreasing their numbers by the same mechanism.

Furthermore, our results also show significantly elevated IL-6 and TNF-α concentrations in AMO-treated rats compared to the control and SOD groups, however significantly lower concentration of IL in the AOM-treated group compared to control and SOD groups.

Inflammatory cytokines such as IL-1α, IL-6, and TNF-α display high levels in colon cancer and play an important role in the mechanism of carcinogenesis.[53],[54] IL-6 and TNF-α are produced by activated macrophages and are potentially involved in the mechanism of cancer development.[54] TNF-α and IL-1α have been shown to be potent markers to be elevated in MAM-induced colon carcinogenesis,[55],[56] which showed an increase in the production of inflammatory cytokines and chemokines during inflammation of the colonic mucosa. Decreased IL-6 and TNF concentrations in GliSODin-treated rats in the colon indicate that colonic mucosa was able to recover after 12 weeks. This suggests that the antioxidant effect of GliSODin goes hand in hand with its anti-inflammatory effect.

Indeed, several studies have shown that plant extracts can exert significant effects, lower the incidence of ACF in the colon, and have the ability to control cancer cellsin vivo and invitro.[57] Among these plant extracts, one could mention that dried beans (Phaseolus spp. Lam);[42] fruits;[58] dietary onion;[59] polyphenol red wine;[54] polyethylene glycol;[60] extract from ginkgo leaves (Ginkgo biloba);[61] polyphenols of tea;[62]Phaleria macrocarpa;[37] carvacrol;[63]Olea europaea L.;[64]Moringa oleifera L.;[65]Dendrophthoe pentandra (L);[66]Curcuma longa;[67] and Strobilanthes crispus[3],[68] demonstrated that natural extracts are protective in colon cancer induced by AOM.

CEA is a generally measured tumor marker in colorectal cancer.[69] In this study, the level of plasma CEA significantly increased in AOM-treated rats. However, rats with GliSODin normalized the level of CEA in plasma. CEA is known to play an important role in colon cancer and tumor invasion and metastasis due to its function as a promoter of cellular aggregation, regulator of the innate immune system, and mediator of signal transduction.[70],[71]

 > Conclusion Top

GliSODin exhibits an anticarcinogenic effect against AOM-induced cytotoxicity in rat colon. The observed chemopreventive effect of these natural extracts was attributed to its antioxidant effects and may be related to its possible anti-inflammatory activity, which merits further study.


The authors of this article would like to thank Dr. Cory Goldberg and Dr. Laurent Intes for their scientific comments and linguistic assistance.

Financial support and sponsorship


Conflicts of interest

There are no conflicts of interest.

 > References Top

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  [Figure 1], [Figure 2], [Figure 3], [Figure 4]

  [Table 1], [Table 2], [Table 3]


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