Scholarly article on topic 'Evaluation of cardioprotective activity of Lepidium sativum seed powder in albino rats treated with 5-fluorouracil'

Evaluation of cardioprotective activity of Lepidium sativum seed powder in albino rats treated with 5-fluorouracil Academic research paper on "Chemical sciences"

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Abstract of research paper on Chemical sciences, author of scientific article — Eman Taha Mohamed, Ghada Mohamed Safwat

Abstract 5-fluorouracil (5-FU) is a chemotherapeutic agent used for treatment of solid tumors. Cardiotoxicity is a major complication of 5-FU therapy. Therefore, the aim of the present study was to evaluate the possible cardioprotective potency of Lepidium sativum seed powder (LS) against 5-FU-induced cardiotoxicity and oxidative stress in albino rats. The rats were divided into three groups. Rats in the control group received saline daily for 8 days only. Rats in FU-treated group received saline orally for 8 days, then I.P. injected with 5-FU (150 mg\kg B.W) on the 5th day. Rats in LS-treated group were orally dosed with LS (550 mg\kg B.W\day) for 8 days, and on the 5th day rats were administrated with the same previous dose of 5-FU. 5-FU induced cardiotoxicity was assessed by a significant increase in serum concentrations of cTnI, CK-MB, lipid profile and a moderate elevation of cardiac MDA. 5-FU significantly decreased serum HDL-c and GSH concentration in cardiac homogenate. Its administration also resulted in the release of some inflammatory markers such as myeloperoxidase and Interleukin-1β (IL-1β). All 5-FU altered parameters were markedly ameliorated by LS pre-co-post-treatment. Results of the present study suggest that LS has a significant effect on the protection of the heart against 5-FU-induced cardiotoxicity through maintaining the antioxidant and anti-inflammatory activities.

Academic research paper on topic "Evaluation of cardioprotective activity of Lepidium sativum seed powder in albino rats treated with 5-fluorouracil"


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Evaluation of cardioprotective activity of Lepidium sativum seed powder in albino rats treated with 5-fluorouracil

Eman Taha Mohamed *, Ghada Mohamed Safwat

Biochemistry Department, Faculty of Veterinary Medicine, Beni-Suef University, Beni-Suef, Egypt


Article history:

Received 5 March 2016

Received in revised form 1 May 2016

Accepted 4 May 2016

Available online


Lepidium sativum



Oxidative stress


5-fluorouracil (5-FU) is a chemotherapeutic agent used for treatment of solid tumors. Cardiotoxicity is a major complication of 5-FU therapy. Therefore, the aim of the present study was to evaluate the possible cardioprotective potency of Lepidium sativum seed powder (LS) against 5-FU-induced cardiotoxicity and oxidative stress in albino rats. The rats were divided into three groups. Rats in the control group received saline daily for 8 days only. Rats in FU-treated group received saline orally for 8 days, then I.P. injected with 5-FU (150 mg\kg B.W) on the 5th day. Rats in LS-treated group were orally dosed with LS (550 mg\kg B.W\day) for 8 days, and on the 5th day rats were administrated with the same previous dose of 5-FU. 5-FU induced cardiotoxicity was assessed by a significant increase in serum concentrations of cTnI, CK-MB, lipid profile and a moderate elevation of cardiac MDA. 5-FU significantly decreased serum HDL-c and GSH concentration in cardiac homog-enate. Its administration also resulted in the release of some inflammatory markers such as myeloperoxidase and Interleukin-1|3 (IL-1|3). All 5-FU altered parameters were markedly ameliorated by LS pre-co-post-treatment. Results of the present study suggest that LS has a significant effect on the protection of the heart against 5-FU-induced cardiotoxicity through maintaining the antioxidant and anti-inflammatory activities.

© 2016 Beni-Suef University. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (


1. Introduction

5-fluorouracil (5-FU) is an antimetabolite fluoropyrimidine analog of the nucleoside pyrimidine with antitumor activity. 5-FU is widely used systemically for breast, gastrointestinal, pancreatic and skin cancers (Rossi, 2013). The main mechanism of action is interfering with DNA synthesis and mRNA

transcription. However, cardiotoxicity is one of the most important associated side effects resulting from its nonspecific cytotoxicity in cancer cells (Alvarez et al., 2012; Carrillo et al., 2015). Clinical cardiac toxicities associated with intravenous infusion of 5-FU covers a wide range of manifestations like angina, myocardial ischemia, congestive heart failure, myo-cardial infarction, pulmonary edema, and sudden death (Polk et al., 2014; Sorrentino et al., 2012; Tsavaris et al., 2002). Recent

* Corresponding author. Biochemistry Department, Faculty ofVeterinary Medicine, Beni-Suef University, Beni-Suef, Egypt. Tel.: +20 822327982; fax: +20 822327982.

E-mail address: (E.T. Mohamed).

2314-8535/© 2016 Beni-Suef University. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (


studies showed that the incidence of 5-FU-induced cardiotoxicity was in the range of 0-20% (Polk et al., 2014), and highlighted an increased risk of toxicity during the administration of higher doses of 5-FU with continuous infusion and not bolus administration. After administration, 5-FU follows different metabolic fates; more than 80% of the dose is inactivated by hepatic biotransformation, about 15-20% is excreted in the urine and only a small fraction remains available to exert its antineoplastic activity (Casale et al., 2004).

Different mechanisms of 5-FU induced cardiotoxicity are proposed, including direct drug or drug metabolite-mediated toxic action on myocytes (Mizuno et al., 1995), coronary vasospasm and thrombogenic effects (Bertolini et al., 2001). 5-FU induces the endothelial damage and extravasation of blood with the drug into cardiac tissue resulting in an inflammatory reaction and myofibril necrosis (Bertolini et al., 2001; Kumar et al., 1995). Another theory has suggested cardiotoxic impurities in the 5-FU formulation (fluoroacetaldehyde, generated in the alkaline solution of fluorouracil during storage, which may be converted to a cardiotoxic agent, fluoroacetate) (Arellano et al., 1998; Becker et al., 1999). Fluoroacetate enters the Krebs' cycle and converts into fluorocitrate, which inhibits the enzyme aconitase (Keller et al., 1996) causing citrate accumulation, disruption of the tricarboxylic acid cycle and severe impairment of energy production within the myocytes (Gradishar and Vokes, 1990). The pathogenesis of 5-FU induced cardiotoxicity may involve cellular damage due to the oxidative stress and the induction of apoptosis (Rashid et al., 2014). Accordingly, therapeutic interventions having antioxidant activity may be effective against oxidative stress associated with cardiovascular diseases.

Lepidium sativum (LS) is locally known as 'hub arachad' belonging to family Brassiaceae. Chemically, the plant seeds and leaves contain flavonoids and isothiocynates glycosides, essential oils, carbohydrates, proteins, fatty acids, p-carotene and vitamins like riboflavin, niacin and ascorbic acid (Yadav et al., 2010). The seeds are consumed in salad and as a spice (Maier et al., 1998). The plant leaves and seeds have been used in traditional medicine. The plant is reported to have hypoglycemic, antihypertensive, diuretic (Jouad et al., 2001; Patel et al., 2009) hemagglutinating, fracture healing (Eddouks et al., 2005; Yadav et al., 2011), anti-inflammatory (Raval et al., 2013), hepato-protective (Abuelgasim et al., 2008; Al-Asmari et al., 2015), an-tioxidant (Agarwal and Verma, 2011; Zia-Ul-Haq et al., 2012) and anti-carcinogenic activities (Maghrani et al., 2005).

The present study was designed to investigate the cardio-protective effect of LS against 5-FU-induced cardiotoxicity in rats by studying some biochemical cardiac injury markers, an-tioxidant defense system, serum lipid profile and inflammatory markers.

2. Materials and methods

2.1. Chemicals

5-fluorouracil was obtained from ACDIMA International (AiT) Shanghai Xudong Haipu Pharmaceutical Co., Ltd. Reduced glutathione (GSH), malondialdehyde (MDA) and nitric oxide (NO) commercial kits were purchased from Bio-diagnostic Company

for research kits, Egypt. Triacylglycerol (TAG), total cholesterol (TC) and HDL-cholesterol commercial diagnostic kits were purchased from Spinreact Company, Spain. Rat Troponin I (cTnl) (Catalog number KT-639), rat Creatinine kinase-MB isoenzyme (CK-MB) (Catalog number KT-12247) and rat myeloperoxidase (Catalog number KT-60345) immunoassay kits were purchased from Kamiya Biomedical Company, USA. Rat Interleukin-1p (IL-1p) (Catalog number K 0331212) ELISA kit was purchased from Komabiotech Company, Korea.

2.2. Plant material

Lepidium sativum L. seeds (known as garden cress) (LS) were obtained from Agricultural Research Center, Egypt. Garden cress seeds were dried and ground to powder. The suspension of Lepidium seed powder was made with a sufficient quantity of distilled water by a dose of 550 mg/kg B.W (Raval and Ravishankar, 2010). It was administered through the oral route with the help of a gastric gavage. Other non-mentioned chemicals used in the present study were of the highest analytic grade and purchased from Sigma-Aldrich Company, USA.

2.3. Animals and experimental design

Thirty adult male albino Sprague-Dawley rats (120-150g) were provided by the Helwan farm of laboratory animals Cairo, Egypt. Rats were kept in standard cages at room temperature (25 ± 2 °C) with a 12:12 h dark-light cycle and humidity (70%). All animals were allowed free access to water and fed with uniformly basal diet. All experimental procedures were conducted in accordance with the guide for the care and use of laboratory animals and in accordance with the local Animal Care and Use Committee. After acclimatization, rats were randomly divided into 3 groups (n = 10). Animals in the control group received only saline daily for 8 days by oral gavage. Animals in FU-treated group received saline orally for 8 days, then were given a single dose of 5-FU (150 mg\kg B.W\I.P injection on the 5th day) (Blijham, 1991). Animals in LS-treated group received a suspension of LS seed powder (550 mg\kg\day) (Raval and Ravishankar, 2010) orally for 8 days and were injected with a single dose of 5-FU (150 mg\kg B.W\I.P) on the 5th day.

2.4. Blood sampling and tissue preparation

Blood samples were collected 24 hours after the last dose, and all rats were sacrificed by cervical decapitation. The obtained sera were monitored for lipid profile, cTnl and CK-MB activity. Heart tissues were excised after dissection of the animals and designated for biochemical analysis. The excised heart tissue (0.5 g) was homogenized in ten volumes of ice cold phosphate buffer (pH:7) until a uniform suspension was obtained. The homogenate was then centrifuged at 20,000 x g for 10 min at 4 °C using high speed cooling centrifuge. The clear supernatant was used for the assay of GSH, MDA, NO, MPO and IL-1p.

2.5. Biochemical assays

2.5.1. Serum analysis

Serum cardiac markers (CK-MB and cTnI) were measured according to instruction of diagnostic kits. Serum lipid profile (TAG,


beni-suef university journal of basic and applied sciences


TC and HDL-cholesterol) were assayed spectrophotometri-cally with the enzymatic colorimetric method described by authors (Burstein et al., 1970; Fossati and Principe, 1982; Richmond, 1973), respectively. VLDL-cholesterol was calculated as TG/5 while LDL-cholesterol was calculated by the formula [LDL-c = total cholesterol-HDL-c-VLDL-c] according to Fruchart (1982). Atherogenic index was calculated by the equation (LDL-c/HDL-c) described by Castelli and Levitar (1977).

2.5.2. Heart tissue analysis

Oxidant\antioxidant status in heart tissues includes GSH concentration was determined according to the method of Beutler et al. (1963). Lipid peroxides as MDA concentration were measured according to the method of Satoh (1978). NO concentration was determined by using biochemical method of Montgomery and Dymock (1961). Pro-inflammatory cytokines in cardiac tissue (MPO and IL-1B) were estimated according to instruction of diagnostic kits.

2.6. Statistical analysis

Statistical analysis was carried out using Graph Pad In stat software (version 3, ISS-Rome, Italy). One way analysis of variance (ANOVA) test followed by Tukey-Kramer (TK) multiple comparison post test were used. The values are expressed as mean ± standard error (SE). The p values below 0.05 were considered statistically significant.

3. Results

Results of cardiac biomarkers were reported in Figs. 1 and 2 and showed a significant increase in serum concentrations of


Control FU-treated LS-treated


50 40 30 20 10

Control FU-treated LS-treated


Fig. 2 - Serum CK-MB activity in different treated groups.

cTnl (Fig. 1) and CK-MB (Fig. 2) in FU-treated group in comparison to control group. This elevation was significantly decreased in LS-treated group, indicating the cardioprotective role of the plant.

Table 1 represented the effects of 5-FU and LS treatment on serum lipid profile, including TAG, TC, HDL-c, VLDL-c LDL-c concentrations in different rat groups. 5-FU treatment significantly increased the serum TAG and TC levels in FU-treated group in comparison to control group indicating hypertriglyceridemia and hypercholesterolemia. Serum LDL-c and VLDL-c concentrations are significantly increased while the serum HDL-c concentration is significantly decreased in FU-treated group in comparison to control group. Pre-co-post-treatment with LS significantly improved the tested parameters. Fig. 3 illustrated the mean ratio of the atherogenic index (LDL-C/HDL-C) in different treated groups. Atherogenic index is a stronger risk predictor of cardiovascular. Results showed a significant increase in this ratio in the FU-treated group which was significantly decreased by LS treatment.

Table 2 represented the concentrations of cardiac homog-enate MPO, IL-1p, GSH, MDA and NO in different groups. It showed a significant increase in inflammatory markers such as myocardial IL-1p and MPO activity and a significant decrease in GSH concentration in the FU-treated group when compared to control rats. LS treatment reversed the results of these tested parameters. However, cardiac MDA and NO concentrations were non-significantly increased in the FU-treated group in comparison to control group and were non-significantly decreased in the LS-treated group.


Fig. 1 - Serum troponin I (cTnl) concentration in different treated groups.

5-fluorouracil is a commonly prescribed chemotherapy for a wide range of solid tumors. When elemental fluorine is reacted


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Table 1 - Serum TAG, cholesterol, HDL-c, VLDL-c, LDL-c concentrations in different treated groups.

TAG (mg/dl) Cholesterol (mg/dl) VLDL-c (mg/dl) HDL-c (mg/dl) LDL-c (mg/dl)

Control 64.39 ± 4.99a 95.05 ± 5.09a 12.88 ± 1.01a 47.62 ± 2.60a 34.55 ± 2.64a

FU-treated 145.2 ± 8.59b 214.1 ± 17.12b 29.04 ± 1.72b 25.43 ± 1.65b 159.7 ± 13.15b

LS-treated 97.14 ± 4.54c 136.2 ± 8.68a 19.43 ± 0.91c 41.77 ± 2.51a 75.01 ± 6.27c

Means with different superscript letters are significantly different at p < 0.05. The data are presented as means ± S.E.

Table 2 - Cardiac concentrations of MPO, IL-1 p, GSH, MDA and NO in different treated groups.

MPO (u/100 mg tissue) IL-1ß (pg/mg ) GSH (mg/g tissue) MDA (nmol/g tissue) NO (| mol/L)

Control 0.410 ± 0.064a 44.60 ± 0.95a 40.24 ± 3.10a 89.70 ± 6.54 41.15 ± 4.27

FU-treated 1.503 ± 0.169b 82.90 ± 0.81b 23.12 ± 2.04b 104.3 ± 6.74 48.50 ± 3.66

LS-treated 0.817 ± 0.023a 50.40 ± 2.15a 33.27 ± 1.50a 93.40 ± 3.81 42.18 ± 3.61

Means with superscript letters are significantly different at p < 0.05. The data are presented as means ± S.E.

Control FU-treated LS-treated Groups

Fig. 3 - The mean ratio of the atherogenic index (LDL-c/ HDL-c) in different treated groups.

with uracil, 5-fluorouracil is produced. Fluorouracil and its metabolites have a number of various mechanisms of action. It acts principally as a thymidylate synthase (TS) inhibitor. Following its administration, 5-FU is rapidly metabolized to 5-fluordeoxyuridine 5'monophosphate (FdUMP) which competitively binds to TS and blocks the methylation of uracil toward thymine which is required for DNA replication. So the cancerous cells undergo cell death via thymine-less death (Longley et al., 2003). Moreover, 5-FU is phosphorylated to triphosphate (F-UTP) and incorporated into RNA instead of uracil, thus blocking their transcription and stopping the growth of cancerous cells. Cytotoxic effects are also deriving from incorporation of FdUTP into DNA as well as F-UTP and 5-fluorocytosine into RNA. These metabolites are thought to influence calcium channel dependent membrane function, to interfere with mitochondrial phosphate metabolism, to alter contractile proteins, to cause oxidative damage and release of vasoactive substances like histamine and catecholamines, and lead to autoimmune mechanisms (Cianci et al., 2003).

The mechanisms of 5-FU-induced cardiotoxicity are hem-orrhagic infarction, myocardial inflammatory reaction with interstitial fibrosis; arterial endothelial injury followed by thrombosis (Bertolini et al., 2001); increased metabolism leading to depletion of ATP, increased levels of superoxide anion and a decreased antioxidant capacity; arterial vasoconstriction and altered plasma levels of substances involved in coagulation and fibrinolysis (Dechant et al., 2012). Oxidative stress causes cellular damage, coronary artery spasm and the decreased affinity of RBCs to transfer oxygen, resulting in myocardial ischemia, cardiac arrest and sudden death (Polk et al., 2014).

Myocardium contains high concentrations of diagnostic markers for myocardial infarction and once metabolically damaged, it releases its content into the extracellular fluid (Farvin et al., 2004; Upaganlawar et al., 2009). Serum creatinine kinase (CK) and troponins are some of these markers. Assay of the CK-MB isoenzyme activity in serum is an important diagnostic indicator owing to its marked excess in myocardial tissue and its consequent sensitivity. The increased activity of serum CK-MB isoenzyme reflects the alterations in the plasma membrane integrity and permeability (Farvin et al., 2004). Cardiac troponin I (cTnl) is a cell-structural protein specific to myocardial tissue. cTnI is sensitive and specific biochemical marker of myocardial cell necrosis and has been considered as the gold standard marker for acute myo-cardial infarction and drug-induced cardiotoxicity (Gaze and Collinson, 2005). In the present study, 5-FU treated rats showed a significant elevation in the activity of serum CK-MB and cTnI level (Figs. 1 and 2), which indicated 5-FU induced myocar-dial necrotic damage and the leakiness of the plasma membrane. LS treatment resulted in lower activity of the CK-MB and cTnl level in serum. It was demonstrated that LS could maintain membrane integrity, thereby limiting the leakage of these biomarkers.

Hyperlipidemia plays an important role in cardiovascular diseases and the development of atherosclerosis (Hassarajani et al., 2007; Neil et al., 1990). A significant elevation in the serum TC, TAG, VLDL-c and LDL-c fractions along with a decrease in HDL-c were observed in 5-FU treated rats compared to control rats (Table 1). These observed changes concerning lipid profile come in agreement with Abdel-Hamid et al. (2011) and could be attributed to the enhanced lipid synthesis via cardiac cyclic


adenosine monophosphate (Paritha and Devi, 1997). Hyper-cholesterolemia and hypertriglyceridemia are mainly related to cardiac ischemia (Jackson and Beaglehole, 1995). The ath-erogenic effect of 5-FU as the mean ratio of LDL-c/HDL-c was significantly increased in FU-treated rats (Fig. 3).The higher atherogenic index, the higher is the risk of the cardiovascular disease (Karthikeyan et al., 2007). Atherogenic index indicates the deposition of foam cells or fatty infiltration of lipids in heart, coronaries, aorta, liver and kidneys.

The treatment with LS successfully restored the altered serum parameters as shown in Table 1. Halaby et al. (2015) reported that a diet supplemented with 5% and 10% LS seed powder improved the altered lipid profile in cisplatin-injected rats. Moreover, treatment with flavonoid and sapogenin extracts of LS lowered TC, TAG, LDL-c and AI values in hyper-lipidemic rats (Shukla et al., 2015). Also, hypolipidemic and hypocholesterolemic effects of LS were mentioned by Umesha and Naidu (2012) and Althnaian (2014).The hypolipidemic effect of LS might be due to inhibition of absorption and enhanced excretion of lipids through the gastrointestinal tract (Chauhan et al., 2012). The hypocholesterolemic effect of LS might be attributed to inhibition of cholesterol biosynthesis via inhibition of HMG-CoA reductase, the rate-limiting enzyme that mediates the first step in cholesterol biosynthesis (Althnaian, 2014). The increased levels of the cardio protective lipoprotein; HDL-c after administration of LS concluded that the plant has a potent cardioprotective effect, and this effect may be due to the increased activity of lecithin: cholesterol acyl transferase (LCAT), an enzyme which plays a key role in incorporating the free cholesterol into HDL-c which is then catabolized by the hepatocytes (Shukla et al., 2015).

Oxidative stress is an overproduction of reactive oxygen species (ROS) with a deficiency of enzymatic and non-enzymatic antioxidants (Valko et al., 2007). Recent studies have suggested that the 5-FU-induced cytotoxicity is linked to the enhanced ROS formation as 5-FU increased intracellular levels of superoxide anion (O-2) (Afzal et al., 2012). As a consequence, the triggering of apoptotic program and cardiomyocyte damage were occurring (Lamberti et al., 2012). However, the exact molecular mechanisms of 5-FU cardiotoxicity have not yet been completely understood. These ROS may attack any type of molecules, but their main target appears to be polyenoic fatty acids within membranes forming peroxyl radicals. These radicals then attack adjacent fatty acids within membranes, causing a chain reaction of lipid peroxidation (Priscilla and Prince, 2009) and myocardial necrosis (Rajadurai and Prince, 2006). MDA is used as a marker of lipid peroxidation (Nielsen et al., 1997). It was elevated in the hearts of guinea pig (Durak et al., 2000) and rat (Eskandari et al., 2014) after 5-FU-treatment. Moreover, Kinhult et al. (2003) suggested that the arterial en-dothelial damage resulted from 5-FU therapy may be due to the generation of ROS. These findings agreed with our results as shown in Table 2 and indicate the occurrence of some degrees of oxidative stress and myocardial damage during 5-FU treatment.

Glutathione depletion is an indicator of oxidative stress. GSH, a tripeptide, is one of the cellular non-enzymatic antioxidant biomolecules. GSH has a direct antioxidant action by reacting with superoxide radicals, peroxy radicals and singlet oxygen followed by the formation of oxidized GSSG and other disulfides.

In addition, it has a role in drug detoxification (Meister, 1988). When isolated cardiomyocytes were incubated with 5-FU, glutathione depletion was progressing as a consequence of ROS formation (Eskandari et al., 2014). In our experiment, the cardiac GSH level was significantly decreased as shown in Table 2 after 5-FU administration. Hence, it is confirmed that the cytotoxic mechanism of 5-FU is mediated via oxidative stress. LS has an antioxidant effect (Halaby et al., 2015; Olorunnisola et al., 2012) so it caused a significant elevation of the GSH level and a nonsignificant decrease of the MDA concentration in cardiac homogenate of LS-treated group. These results were parallel with the data obtained by Behrouzian et al. (2014) and Doke and Guha (2014).

Nitric oxide (NO) is an important signaling messenger known to play important roles in many physiological (such as host defense and homeostasis) and pathological (such as inflammation) conditions (Zhang et al., 2008). In mammalian cells, NO is generated in the cells by the NADPH-dependent oxidation of arginine to citrulline by the enzyme nitric oxide synthase (NOS) (Laskin et al., 1994). Overflow of NO may contribute to inflammatory reaction through nitrosation, oxidative damage, and enhanced release of inflammatory cytokines (Kanwar et al., 2009). Several studies have shown action of 5-FU on the production of NO (Matthews et al., 2001). Jung et al. (2002) suggested the efficacy of 5-FU to block NO production through the inac-tivation of IkB kinase in stomach cancer cells. While, Leitao et al. (2007) suggested an important role of NO in the pathogenesis of oral mucositis induced by 5-FU. In the present study, the 5-FU administration showed no significant change of NO level, which may be attributed to the formation of peroxynitrite by the reaction of NO with generated superoxide radicals (Gandhi et al., 2009).

Myeloperoxidase, an enzyme that is associated with the neu-trophils, has been studied to form an index of the neutrophils infiltration into inflamed tissue. Myeloperoxidase, a green he-moprotein enzyme can use H2O2 generated by NADPH oxidase to oxidize halides (Cl-, Br- and I-) to their corresponding hypohalous acids (an additional class of active oxygen metabolite) (Kettle et al., 1997). MPO generates different reactive oxidants and diffusible radical species that are capable of both initiating lipid peroxidation (Zhang et al., 2002) and stimulating post-translational modifications to target proteins, including halogenation, nitration, and oxidative cross-linking (Heinecke, 2003; Podrez et al., 2000). Consequently, it is linked to both inflammation and oxidative stress. In the present study, the significant increase in myocardial MPO activity is an indicative of 5-FU-induced inflammation of myocardial tissue and neutrophil infiltration. Restored levels of myocardial inflammatory markers in LS-treated group, indicated that the LS suppressed the neutrophil infiltration and inflammatory cytokine release to the injured myocardium.

Interleukin-1ß (IL-1ß) is an important pro-inflammatory cytokines with a relevant role in the inflammatory disorders. IL-1ß is produced by monocytes, macrophages, endothelial cells and fibroblasts (Dung et al., 2009). Okamoto et al. (1998) and Curra et al. (2013) indicated that 5-FU is a potent inducer of several types of cytokines, including IL-1ß.This was achieved in our results reported in Table 2 that showed a significant increase in IL-1ß concentrations in cardiac homogenate of FU-treated rats in comparison to control ones. The increased MPO


and IL-1ß concentrations in cardiac homogenate indicate the inflammatory effect of 5-FU (Soares et al., 2011). LS ameliorated that effect as it significantly decreased the concentration of MPO and IL-1ß in the LS-treated group due to its antiinflammatory activity.

Restored levels of myocardial MDA, GSH, MPO and IL-1ß by LS Administration may be attributed to its high content in antioxidants (vitamin C, E, carotenoids, polyphenols and flavonoids) (Donno et al., 2013; Lee et al., 2013) and its antiinflammatory activities (Calder, 2006; Lopez-Garcia et al., 2004). In this respect, gas chromatography-mass spectrometry analysis of LS showed the presence of alpha-linolenic acid (C18:3) which has an anti-inflammatory activity (Al-Asmari et al., 2015). The role of alpha linolenic acid is to downregulate the gene expression of inflammatory cytokines such as IL-6, IL-1ß and TNF-a (Zhao et al., 2007).

5. Conclusion

Our results confirmed the existence of cardiotoxicity due to 5-FU therapy, which was indicated by an elevation of serum cardiac cTnl, CK-MB, altered lipid profile and atherogenic index with enhanced oxidative stress and the release of some inflammatory markers. It can be concluded that LS seed exerts cardioprotective activity that could be partly contributed by its antioxidant and anti-inflammatory activities. So, Lepidium sativum can be considered a candidate to protect against cardiotoxicity commonly encountered with 5-FU treatment.


We are thankful to university grant commission, Beni Suef University, Beni Suef, Egypt for providing financial support.


Alvarez P, Marchal JA, Boulaiz H, Carrillo E, Velez C, Rodriguez-Serrano F, et al. 5-Fluorouracil derivatives: a patent review. Expert Opin Ther Pat 2012;22(2):107-23.

Abdel-Hamid HF, Soliman A, Helaly FM, Ragab S. Cytotoxic

potency and induced biochemical parameters in mice serum of new furan derivatives against liver cancer cell line. Acta Pol Pharm 2011;68(4):499-505.

Abuelgasim AI, Nuha HS, Mohammed AH. Hepato-protective effect of Lepidium sativum against carbon tetrachloride-induced damage in rats. Res J AniVet Sci 2008;3:20-3.

Afzal S, Jensen SA, Sorensen JB, Henriksen T, Weimann A, Poulsen HE. Oxidative damage to guanine nucleosides following combination chemotherapy with 5 fluorouracil and oxaliplatin. Cancer Chemother Pharmacol 2012;69:301-7.

Agarwal J, Verma DL. Antioxidant activity-guided fractionation of aqueous extracts from Lepidium sativum and identification of active flavonol glycosides. Acad Arena 2011;3:14-18.

Al-Asmari AK, Athara T, Al-Shahranib HM, Al-Dakheelc SI, Al-Ghamdi MA. Efficacy of Lepidium sativum against carbon tetra chloride induced hepatotoxicity and determination of

its bioactive compounds byGC-MS. Toxicol Rep 2015;2:1319-26.

Althnaian T. Influence of dietary supplementation of Garden cress (Lepidium sativum L.) on liver histopathology and serum biochemistry in rats fed high cholesterol diet. J Adv Vet Anim Res 2014;1(4):216-23.

Arellano M, Malet-Martino M, Martino R, Gires P. The anti-cancer drug 5-fluorouracil is metabolized by the isolated perfused rat liver and in rats into highly toxic fluoroacetate. Br J Cancer 1998;77(1):79-86.

Becker K, Erckenbrecht JF, Haussinger D, Frieling T. Cardiotoxicity of the antiproliferative compound fluorouracil. Drugs 1999;57:475-84.

Behrouzian F, Razavi S, Phillips G. Cress seed (Lepidium sativum) mucilage, an overview. Bioact Carbohydr Diet Fibre 2014;doi:10.1016/j.bcdf.2014.01.001.

Bertolini A, Flumano M, Fusco O, Muffatti A, Scarinci A, Pontiggia G, et al. Acute cardiotoxicity during capecitabine treatment: a case report. Tumori 2001;87:200-6.

Beutler E, Duron O, Kelly BM. Improved method for the determination of blood glutathione. J Lab Clin Med 1963;61:882-8.

Blijham GH. Chemotherapy of colorectal cancer. Anticancer Drugs 1991;2:233-45.

Burstein M, Scholnick HR, Morfin R. Rapid method for the

isolation of lipoproteins from human serum by precipitation with polyanions. J Lipid Res 1970;11(583):595.

Calder PC. Polyunsaturated fatty acids and inflammation. Prostaglandins Leukot Essent Fatty Acids 2006;75197-202.

Carrillo E, Navarro SA, Ramirez A, Garcia MA, Grinân-Lisôn C, Perân M, et al. Review: 5-Fluorouracil derivatives: a patent review (2012-2014). Expert Opin Ther Pat 2015;25(10):1131-44.

Casale F, Canaparo R, Serpe L, Muntoni E, Pepa CD, Costa M, et al. Plasma concentrations of 5-fluorouracil and its metabolites in colon cancer patients. Pharmacol Res 2004;50:173-9.

Castelli T, Levitar Y. Atherogenic index. Curr Presc 1977;39.

Chauhan K, Sharma S, Agarwal N, Chauhan S, Chauhan B. A

study on potential hypoglycemic and hypolipidemic effects of Lepidium sativum (Garden cress) in alloxan induced diabetic rats. Am J PharmTech Res 2012;2:522-35.

Cianci G, Morelli MF, Cannita K, Morese R, Ricevuto E, Di Rocco ZC, et al. Prophylactic options in patients with 5-fluorouracil-associated cardiotoxicity. Br J Cancer 2003;88:1507-9.

Curra M, Martins MA, Lauxen IS, Pellicioli AC, Sant'Ana Filho M, Pavesi VC, et al. Effect of topical chamomile on immunohistochemical levels of IL-1ß and TNF-a in 5-fluorouracil-induced oral mucositis in hamsters. Cancer Chemother Pharmacol 2013;71(2):293-9.

Dechant C, Baur M, Böck R, Czejka M, Podczeck-Schweighofer A, Dittrich C, et al. Acute reversible heart failure caused by coronary vasoconstriction due to continuous 5-fluorouracil combination chemotherapy. Case Rep Oncol 2012;5:296-301.

Doke S, Guha M. Garden cress (Lepidium sativum L.) seed - an important medicinal source: a review. Scholars research library. J Nat Prod Plant Resour 2014;4(1):69-80.

Donno D, Beccaro G, Mellano M, Canterino S, Cerutti A, Bounous G. Improving the nutritional value of kiwifruit with the application of agroindustry waste extracts. J Appl Bot Food Qual 2013;86:11-15.

Dung NT, Bajpai VK, Yoon JI, Kang SC. Anti inflammatory effects of essential oil isolated from the buds of Cleistocalyx operculatus (Roxb) Merr and Perry. Food Chem Toxicol 2009;47:449-53.

Durak I, Karaayvaz M, Kavutcu M, Cimen MY, Kacmaz M,

Buyukkocak S, et al. Reduced antioxidant defense capacity in myocardial tissue from guinea pigs treated with 5-fluorouracil. J Toxicol Environ Health A 2000;59:585-9.


Eddouks M, Maghrani M, Zeggwagh NA, Michel JB. J Ethnopharmacol 2005;97(2):391-5.

Eskandari MR, Moghaddam F, Shahraki J, Pourahmad J. A comparison of cardiomyocyte cytotoxic mechanisms for 5-fluorouracil and its pro-drug capecitabine. Xenobiotica 2014;1-9.

Farvin KHS, Anandan R, Kumar SH, Shiny KS, Sankar TV,

Thankappan TK. Effect of squalene on tissue defense system in isoproterenol-induced myocardial infarction in rats. Pharmacol Res 2004;50(3):231-6.

Fossati P, Principe L. Enzymatic colorimetric method to determination triglycerides. Clin Chem 1982;28:2077-80.

Fruchart GG. LDL-cholesterol determination after separation of low density lipoprotein. Rev Fr Des Laboratories 1982;103(7):117.

Gandhi C, Upaganlawar A, Balaraman R. Protection against in vivo focal myocardial ischemia/reperfusion injury induced arrhythmias and apoptosis by Hesperidin. Free Radic Res 2009;43:817-27.

Gaze DC, Collinson PO. Review: cardiac troponins as biomarkers of drug- and toxin-induced cardiac toxicity and cardioprotection. Expert Opin Drug Metab Toxicol 2005;1(4):715-25.

Gradishar WJ, Vokes EE. 5-Fluorouracil cardiotoxicity: a critical review. Ann Oncol 1990;1:409-14.

Halaby MS, Farag MH, Mahmoud SAA. Protective and curative effect of garden cress seeds on acute renal failure in male albino rats. Middle East J Appl Sci 2015;5(2):573-86.

Hassarajani S, Souza TD, Mengi SA. Efficacy study of the

bioactive fraction (F-3) of Acorus calamus in hyperlipidemia. Indian J Pharmacol 2007;39:196-200.

Heinecke JW. Oxidative stress: new approaches to diagnosis and prognosis in atherosclerosis. Am J Cardiol 2003;91:12A-16A.

Jackson R, Beaglehole R. Evidence-based management of dyslipidaemia. Lancet 1995;346:1440-1.

Jouad H, Haloui M, Rhiouani H, El Hilaly J, Eddouks M. J Ethnopharmacol 2001;77:175-82.

Jung ID, Yang SY, Park CG, Lee KB, Kim JS, Lee SY, et al.

5-Fluorouracil inhibits nitric oxide production through the inactivation of IkB kinase in stomach cancer cells. Biochem Pharmacol 2002;64(10):1439-45.

Kanwar JR, Kanwar RK, Burrow H, Baratchi S. Recent advances on the roles of NO in cancer and chronic inflammatory disorders. Curr Med Chem 2009;16:2373-94.

Karthikeyan K, Bai BRS, Devaraj SN. Efficacy of grape seed proanthocyanidins on serum and heart tissue lipids in rats subjected to isoproterenol-induced myocardial injury. Vascul Pharmacol 2007;47:295-301.

Keller DA, Roe DC, Lieder PH. Fluoroacetate-mediated toxicity of fluorinated ethanes. Fundam Appl Toxicol 1996;30:213-19.

Kettle AJ, Dalen J V, Winterbourn CC. Peroxynitrite and myeloperoxidase leave the same footprint in protein nitration. Redox Rep 1997;3:257-8.

Kinhult S, Albertsson M, Eskilsson J, Cwikiel M. Effects of

probucol on endothelial damage by 5-fluorouracil. Acta Oncol 2003;42:304-8.

Kumar S, Gupta RK, Samal N. 5-fluorouracil induced

cardiotoxicity in albino rats. Mater Med Pol 1995;27(2):63-6.

Lamberti M, Porto S, Marra M, Zappavigna S, Grimaldi A, Feola D, et al. 5-fluorouracil induces apoptosis in rat cardiomyocytes through intracellular oxidative stress. J Exp Clin Cancer Res 2012;31:60.

Laskin JD, Heck DE, Laskin DL. Multifunctional role of nitric oxide in inflammation. Trends Endocrinol Metab 1994;5:377-82.

Lee CC, Shen SR, Lai YJ, Wu S-C. Rutin and quercetin, bioactive compounds from tartary buckwheat, prevent liver inflammatory injury. Food Funct 2013;4:794-802.

Leitäo RFC, Ribeiro RA, Bellaguarda EAL, Macedo FDB, Silva LR, Oria RB, et al. Role of nitric oxide on pathogenesis of 5-fluorouracil induced experimental oral mucositis in hamster. Cancer Chemother Pharmacol 2007;59(5):603-12.

Longley DB, Harkin DP, Johnston PG. 5-fluorouracil: mechanisms of action and clinical strategies. Nat Rev Cancer 2003;3(5):330-8.

Lopez-Garcia E, Schulze MB, Manson JE, Meigs JB, Albert CM, Rifai N, et al. Consumption of (n-3) fatty acids is related to plasmabiomarkers of inflammation and endothelial activation in women. J Nutr 2004;134:1806-11.

Maghrani M, Zeggwagh NA, Michel JB, Eddonks MJ. Antihypertensive effect of Lepidium sativum L. in spontaneously hypertensive rats. J Ethnopharmacol 2005;100(1-2):193-7.

Maier UH, Gundlach H, Zenk MH. Phytochemistry 1998;6:1791-5.

Matthews NE, Adams MA, Maxwell LR, Gofton TE, Graham CH. Nitric oxide-mediated regulation of chemosensitivity in cancer cells. J Natl Cancer Inst 2001;93:1879-85.

Meister A. Glutathione metabolism and its selective modification. J Biol Chem 1988;263:7205-8.

Mizuno Y, Hokamura Y, Kimura T, Kimura Y, Kaikita K, Yasue H. A case of 5-fluorouracil cardiotoxicity simulating acute myocardial infarction. Jpn Circ J 1995;59(5):303-7.

Montgomery HAC, Dymock JE. The determination of nitrite in water. Analyst 1961;86:414-16.

Neil HA, Mant D, Jones L, Morgan B, Mann JI. Lipid screening: is it enough to measure total cholesterol concentration? BMJ 1990;301:584-7.

Nielsen F, Mikkelsen BB, Nielsen JB, Andersen HR, Grandjean P. Plasma malondialdehyde as biomarker for oxidative stress: reference interval and effects of life-style factors. Clin Chem 1997;43:1209-14.

Okamoto M, Kasetani H, Kaji R, Goda H, Ohe G, Yoshida H, et al. eis Diamminedichloroplatinum and 5-fluorouracil are potent inducers of the cytokines and natural killer cell activity in vivo and in vitro. Cancer Immunol Immunother 1998;47(4):233-41.

Olorunnisola O, Bradley G, Afolayan A. Protective effect of T. violaeea rhizome extract against hypercholesterolemia-induced oxidative stress in Wistar rats. Molecules 2012;17:6033-45.

Paritha IA, Devi CS. Effect of a-tocopherol on isoproterenol-induced changes in lipid and lipoprotein profile in rats. Ind J Pharmacol 1997;29:399-404.

Patel U, Kulkarni M, Undale V, Bhosale A. Evaluation of diuretic activity of aqueous and methanol extracts of Lepidium sativum garden cress (Cruciferae) in rats. Trop J Pharm Res 2009;8(3):215-19.

Podrez EA, Abu-Soud HM, Hazen SL. Myeloperoxidase-generated oxidants and atherosclerosis. Free Radic Biol Med 2000;28:1717-25.

Polk A, Vistisen K, Vaage-Nilsen M, Nielsen DL. A systematic review of the pathophysiology of 5-fluorouracil-induced cardiotoxicity. BMC Pharmacol Toxicol 2014;15:47.

Priscilla DH, Prince PSM. Cardioprotective effect of gallic acid on cardiac troponin-T, cardiac marker enzymes, lipid peroxidation products and antioxidants in experimentally induced myocardial infarction in Wistar rats. Chem Biol Interact 2009;179:118-24.

Rajadurai M, Prince PSM. Preventive effect of naringin on lipid peroxides and antioxidants in isoproterenol-induced cardiotoxicity in Wistar rats: biochemical and histopathological evidences. Toxicology 2006;228:259-68.

Rashid S, Ali N, Nafees S, Hasan SK, Sultana S. Mitigation of 5-Fluorouracil induced renal toxicity by chrysin via targeting oxidative stress and apoptosis in Wistar rats. Food Chem Toxicol 2014;66:185-93.


beni-suef university journal of basic and applied sciences


Raval ND, Ravishankar B. Analgesic effect of Lepidium sativum Linn. (Chandrashura) in experimental animals. Ayu 2010;31(3):371-3.

Raval ND, Ravishankar B, Ashok BK. Anti-inflammatory effect of Chandrashura (Lepidium sativum Linn.) an experimental study. Ayu 2013;34:302-4.

Richmond W. Preparation and properties of a cholesterol oxidase from Nocardia sp. and its application to the enzymatic assay of total cholesterol in serum. Clin Chem 1973;19(12):1350-6.

Rossi S, editor. Australian medicines handbook. 2013 ed. Adelaide: The Australian Medicines Handbook Unit Trust; 2013 ISBN 978-0-9805790-9-3.

Satoh K. Serum lipid peroxide in cerebrovascular disorders determined by a new colorimetric method. Clin Chim Acta 1978;90(1):37-43.

Shukla AK, Bigoniya P, Soni P. Hypolipidemic activity of Lepidium sativum Linn. seed in rats. IOSR J Pharm Biol Sci 2015;10(4):13-22.

Soares PM, Lima-Junior RC, Mota JM, Justino PF, Brito GA, Ribeiro RA, et al. Role of platelet-activating factor in the pathogenesis of 5-fluorouracil-induced intestinal mucositis in mice. Cancer Chemother Pharmacol 2011;68(3):713-20.

Sorrentino MF, Kim J, Foderaro AE, Truesdell AG. 5-fluorouracil induced cardiotoxicity: review of the literature. Cardiol J 2012;19:453-8.

Tsavaris N, Kosmas C, Vadiaka M, Efremidis M, Zinelis A,

Beldecos D, et al. Cardiotoxicity following different doses and schedules of 5-fluorouracil administration for malignancy- a survey of 427 patients. Med Sci Monit 2002;8(6):PI51-7.

Umesha SS, Naidu KA. Vegetable oil blends with a-linolenic acid rich Garden cress oil modulate lipid metabolism in experimental rats. Food Chem 2012;135(4):2845-51.

Upaganlawar A, Gandhi C, Balaraman R. Effect of green tea and vitamin E combination in isoproterenol induced myocardial infarction in rats. Plant Foods Hum Nutr 2009;64:75-80.

Valko M, Leibfritz D, Moncol J, Cronin MT, Mazur M, Telser J. Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol 2007;39:44-84.

Yadav YC, Srivastav DN, Seth AK, Saini V, Balaraman R, Ghelani TK. In vivo antioxidant potential of Lepidium sativum seeds in albino rats using cisplatin induced nephrotoxicity. Int J Phytomed 2010;2:292-8.

Yadav YC, Jain A, Srivastava DN, Jain A. Fracture healing activity of ethanolic extract of Lepidium sativum L. seeds in internally fixed rats' femoral osteotomy model. Int J Pharm Pharm Sci 2011;3(2):193-7.

Zhang J, Knapton A, Lipshultz SE, Weaver JL, Herman EH.

Isoproterenolinduced cardiotoxicity in Sprague-Dawley rats: correlation of reversible and irreversible myocardial injury with release of cardiac troponin T and roles of iNOS in myocardial injury. Toxicol Pathol 2008;36:277-8.

Zhang R, Brennan ML, Shen Z, MacPherson JC, Schmitt D, Molenda CE, et al. Myeloperoxidase functions as a major enzymatic catalyst for initiation of lipid peroxidation at sites of inflammation. J Biol Chem 2002;277:46116-22.

Zhao TD, Etherton KR, Martin PJ, Gillies SG. Dietary linolenic acid inhibits proinflammatory cytokine production by peripheral blood mononuclear cells in hypercholesterolemic subjects. Am J Clin Nutr 2007;85:385-91.

Zia-Ul-Haq M, Ahmad S, Calani L, Mazzeo T, Del Rio D, Pellegrini N, et al. Compositional study and antioxidant potential of Ipomoea hederacea Jacq and Lepidium sativum L. seeds. Molecules 2012;17:10306-21.