Introducing Cichorium Pumilum as a PotentialTherapeutical Agent Against Drug-Induced BenignBreast Tumor in Rats

M-Ali H. Al-Akhras1, Khaled Aljarrah1, Hasan Al-Khateeb1, Adnan Jaradat1,Abdelkarim Al-omari2, Amjad Al-Nasser3, Majed M. Masadeh4,Amr Amin5, Alaaeldin Hamza5, Karima Mohammed5,Mohammad Al Olama6 & Sayel Daoud7

1Department of Physics, Bio-Medical Physics Laboratory, Jordan University of Science &Technology (JUST), Irbid, Jordan, 2Department of General and Pediatric Surgery,Faculty of Medicine, Jordan University of Science and Technology (JUST), Jordan,3Department of Statistics, Yarmouk University, Irbid, Jordan, 4Department ofPharmaceutical Technology, Faculty of Pharmacy, Jordan University of Science andTechnology (JUST), Jordan, 5Biology Department, UAE University, Al Ain, UAE,6Department of Microbiology, Zayed Complex for Herbal Research and TraditionalMedicine, Abu Dhabi, UAE, and 7Pathology Department, Tawam Hospital, Al Ain, UAE

Cichorium Pumilum (chicory) is could be a promising cancer treatment in which aphotosensitizing drug concentrates in benign tumor cells and activated by quanta at certainwavelength. Such activated extracts could lead to cell death and tumor ablation. Previous studieshave shown that Cichorium Pumilum (chicory) contains photosensitive compounds such ascichoriin, anthocyanins, lactucin, and Lactucopicrin. In the present study, the protective effect ofsun light-activated Cichorium against the dimethylbenz[a]anthracene (DMBA) induced benignbreast tumors to female Sprague-Dawley rats was investigated. Chicory’s extract has significantlyincrease P.carbonyl (PC) and malondialdehyde (MDA) and decreases the hepatic levels of totalantioxidant capacity (TAC) and superoxide dismutase (SOD) in benign breast tumors-inducedgroup compared to control. It also significantly decrease the number of estrogen receptorsER-positive cells in tumor masses. These results suggest that chicory extracts could be used asherbal photosensitizing agent in treating benign breast tumor in rats.

Keywords Chicory, Benign breast tumor, Photohemolysis, Oxidative stress

INTRODUCTION

It is necessary for cells to remain healthy to have an intact antioxidant capability tocompensate for oxidant forces (Delimaris et al., 2007). The susceptibility of tissue tofree radicals attack is function of the overall balance between the magnitude ofoxidative stress and its own antioxidant potential (Stadelmann-Ingrand et al., 2004).Healthy cells have developed an antioxidant system which consists of enzymatic

Address correspondence to M-Ali H. Al-Akhras, Department of Physics, Bio-Medical PhysicsLaboratory, Jordan University of Science & Technology (JUST), P.O. Box:3030, Irbid 22110, Jordan.E-mail: alakmoh@just.edu.jo

Electromagnetic Biology and Medicine, Early Online: 1–11, 2012Copyright Q Informa Healthcare USA, Inc.ISSN: 1536-8378 print / 1536-8386 onlineDOI: 10.3109/15368378.2012.662193

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mechanisms such as superoxide dismutase (SOD), catalase (CAT), and nonenzymatic mechanisms including the reduced glutathione (Bourre et al., 1993).

Photodynamic therapy (PDT) involves administration of tumor localizingphotosensitizer agent that produces reactive oxygen radicals during light irradiationand ultimately leads to cell death (Petrovic et al., 2004; Shindo et al., 1993). Thereare two well-defined mechanisms for generating cytotoxic species: the firstmechanism produces free radicals or superoxide ions resulting from hydrogenor electron transfer; second mechanism is singlet oxygen (1O2) which generatedvia an energy transfer process that occurs during collision of excited sensitizerwith oxygen.

The relationship between cancer and UV-induced oxidative damage has beenreported (Kripke, 1981). UV-induced DNA damage can also inhibit DNA repair (Nishiet al., 1981). Sunlight, which consists of broad spectrum of UV, can be used as aphoto-source to treat cancer through a photochemical reaction with previouslyinjected photosensitizers that could cause tumor necrosis. UV incorporated withreactive oxygen species (ROS), such as superoxide anion (O21 or · 2 ), hydroxylradical (OH 2 ), and hydrogen peroxide (H2O2), could cause oxidative damage(Wang et al., 1992). ROS was shown to peroxidate the membrane lipids and induceDNA-protein cross linking, pyrimidine dimmers, and single-strand breaks(Hanimoglu et al., 2007). Also, it was shown that ROS was involved in all steps oftumorigensis, from cancer development to malignant conversion (Ray et al., 2000).Clinical investigations and epidemiological reports have proven the role of ROS in theetiology of cancer (Al-Akhras, 2006b).

Many photosensitizers such as Photofrin, Hypericin, Lutetium Lexaphyrin,Protoporphyrin IX are already known and some of them are used in vivo (Al-Akhras,2006a; Al-Akhras et al., 2007a; Al-Akhras and Grossweiner, 1996; Bilgin et al., 2000).The need to search for photosensitizers that have better quantum efficiency,reductions in toxicity and ability to enable targeting of highly effective payloadremains a desirable therapeutic goal.

Recently, Cichorium Pumilum Jacq (chicory) has been shown to have a strongabsorbance in the UV region from 210–360 nm (Bischoff et al., 2004). Crude extractsfrom flowers and aerial parts of Cichorium Pumilum contain more than onephotosensitive compound; flowers extracts contain cichoriin (Shindo et al., 1993),root extracts contain Lactucin and Lactucopicrin (Norbaek et al., 2002), andanthocyanins (Al-Akhras et al., 2007b). Therefore, Cichorium is expected to be apromising herbal agent due to its strong photosensitivity and temperature dependent(Bischoff et al., 2004; Giese, 1980). Same as other herbs, such as Hypericum which isresponsible for the photodynamic diseases of grazing animals (Pan et al., 2003),Cichorium could be responsible for the sudden death of white sheeps after grazing onchicory and exposed to light (Bischoff et al., 2004).

The main objective of the present study is to assess the potential of chicory’sextract as an efficient agent with solar exposure dose against drug-induced benignbreast tumor in rats.

MATERIALS AND METHODS

ChemicalsSerum aspartate aminotransferase activity was purchased from BioMerieux, RCSLyon, France. O-Dianisidine, 2,4-dinitophenylhydrazine, Thiobarbituric acid, Folinreagent, Epinephrine, SOD enzyme, H2O2, and bovine albumin were obtained fromSigma Chemical Co., St. Louis, MO, USA. All other chemicals were obtained fromlocal commercial suppliers.

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Plant ExtractChicory was collected from a local wild area in northern Jordan. Collected chicoryflower and aerial parts were washed with running water and air dried in a dark room,and afterwards they were cut into small pieces then grinded for a short period of timeto get powder mixtures. Mixtures were extracted by microwave-assisted extractionmethod (Bischoff et al., 2004; Giese, 1980; Hazra et al., 2002). This method is based onheat selection for a solution of 50% ethanol and 50% water and contains the targetextract. This process allows heat selection for the compound without excessiveenergy time consuming.

Hundred grams of dried powder were mixed in 1,000ml of 50% ethanol and 50%water (1 gm/10ml). The suspension was divided into four 250-ml conical flask.Mixtures were then irradiated with a 300-W microwave for two minutes, and wasirradiated for 25 s with power on to give the desired temperature of about 85oC andthen disconnect immediately and re-irradiate for 5 s with power on for heating andthen for 10min with power off for cooling. The process was repeated three timeswithout reaching the boiling condition. The suspension kept over night inside therefrigerator. The extract was finally filtered through gauze and evaporated usinga rotary evaporator under vacuum at 408C. The herbal extraction then kept in therefrigerator for further use.

Animals and Experimental ProtocolFemale Sprague-Dawley rats (150–200 g body weight) were used in theseexperiments. They were fed standard pellets diet and tap water ad libitum, placedin polycarbonate cages with wood chip bedding under a 12 h light/dark cycle, andkept at room temperature of 21 ^ 18C. Rats were acclimatized prior to experimentalby one week. Then at day 50 postpartum, they were randomly divided into6 individual groups each of n ¼ 8: the first group served as sham-control, receiveda daily dose of distilled water (W) based on body weight to an equivalent volumesame as treated animals; the second group received herbal extract (H) and exposed todirect sun light (L); the third group who received a single carcinogenic dose ofdimethylbenz[a]anthracene (DMBA) to induce benign breast tumor (BBT) aretreated withW only; the fourth group BBT and H only; the fifth group, BBT and L; andthe sixth group treated with BBT plus H and L.

The carcinogenic dose of DMBA (50mg/kg body) was based on previous cancerstudies in rodents (Costa et al., 2002). The DMBA was administered by gavage toinduce BBT in female Sprague-Dawley rats. Animals which received light were dailyexposed for 15min during the experiment time. The light exposure dose was(720–1500W/cm2). The crude extract of herb was given orally at daily doses of0.5 g/kg b.wt (Sorg, 2004). At the end of 120 day-experimental period, rats wereanesthetized with diethyl ether and sacrificed by decapitation. Animals were starvedovernight before sacrifice. All experiments were performed in accordance withprotocols approved by the local experimental ethics committee at faculty ofmedicine, Jordan University of Science & Technology (JUST). Procedures for the careand use of research animals at JUST meet the international laws and regulations.

Samples PreparationBlood samples were collected from the retro-orbital plexus of the anesthetized ratsand mammalian glands were removed. The rat’s BBT were performed on section ofstained with hematoxylin and eosin for histopathological examination. Liver sampleswere immediately fixed in 10% buffered formalin. For biochemical studies, the rightlobe of the liver was homogenized in ice-cold KCl (150mmol/l). The ratio of tissueweight to homogenization buffer was 1:10. Suitable dilutions were prepared to

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determine the levels of calcium, protein carbonyl (PC), malondialdehyde (MDA), andtotal antioxidant capacity (TAC). Dilutions with different buffers were used to assessthe activity of SOD) and CAT.

Histopathology and Biochemistry AssayThe most prominent product of lipid peroxidation (LP) is MDA which is used as anindirect index of LP in biological system (Mihara and M. Uchiyama, 1978). Themethod of Aebi (1984) was used to determine MDA which is based on its reactionwith thiobarbituric acid (TBA) to form a pink complex with absorption maximumpeak at 535 nm.

CAT activity was determined bymeasuring the exponential disappearance of H2O2at 240nm and expressed in units/mg of protein as described by Sun and Zigman(1978). The SOD enzyme activity was determined according to the method describedby Reznick et al. (1994). This method is based on the ability of SOD to inhibit the auto-oxidation of epinephrine at alkaline pH to adrenochrome and other derivatives, whichare easily monitored in the near-UV region of the absorption spectrum.

PC contents were determined according to the method of Benzie and Strain(1996). This method is based on spectrophotometric detection of the end product ofthe reaction of 2,4-dinitophenylhydrazine with PC to form protein hydrazones at370 nm. The results were expressed as nanomoles of carbonyl group per milligram ofprotein with molar extinction coefficient of 22,000 l mol21 cm21.

The TAC in liver tissues was evaluated with the ferric reducing antioxidant power(FRAP) assay. The FRAP assay was determined according to the method described byLowry et al. (1951). The FRAP assay measures the change in absorbance at 593 nmcaused by the formation of a blue-colored ferrous-tripyridyltriazine complex from aninitially colorless oxidized ferric form. Tissue-embedded electron-donatingantioxidants drive the reduction of Fe (III) to Fe (II).

The total protein contents of liver tissues were determined according to the Lowrymethod (Peterson, 1977) as modified by Ian and Macdonald (2001). Absorbance wasrecorded using a Shimadzu recording spectrophotometer (UV-160) for allmeasurements. For the histological examinations, small pieces of heart tissue werefixed in 10% neutral phosphate-buffered formalin. Hydrated tissue sections, 5mm inthickness, were then stained with hematoxylin and eosin. The sections wereexamined under a Leica DMRB/E light microscope.

Statistical AnalysisSPSS (version 15) (SPSS Inc., Chicago, IL, USA) was used to assess the statisticaldifference between the groups. To determine a suitable test, the data first werechecked for normality assumption. Because the sample size is small for all groups,Shapiro-Wilk test was used. ANOVA (Parametric) is used if the normality assumption issatisfied or Kruskal Wallis H-Test (non parametric) if normality assumption notsatisfied. If the result of H-Test indicates that there is statistical difference betweengroups, a pairwise comparison test were performed using Mann Whitney U- Test. Ifresults of ANOVA indicate that there is statistical difference between groups, amultiplecomparison test (Exact Fisher LSD test) is used for pair wise comparisons. Data wasexpressed as means ^SD. Significant differences were considered at p , 0.05.

RESULTS

Histological evaluationNo observed acute toxicity after the administration of DMBA or the herb was noticed.In the present study, DMBA-induced typical mammary lobular hyperplasia was

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histopathologically diagnosed. Mammary gland pathology treated with DMBA wasclassified into ductal or lobular hyperplasia and fibroadenoma. These lesions wereconsidered as benign because the basal layer is maintained and the nuclei areuniform without hyperchromasia. Ductal or lobular hyperplasia was observed in allrats treated with DMBA and was significantly higher than those of control group aswell as group treated with Cichorium and light. Concomitant treatment with DMBACichorium and light treated rats are significantly decreased the percentage of ductalor lobular hyperplasia (Fig. 1).

Effects on Estrogen Receptors ExpressionEstrogen receptors (ER) distributions of all tested groups are shown in Fig. 2D–F.Positive ER-immunoreactive had a strong dark brown stain within the nuclei of theepithelial cells of the mammary ducts. Samples from sham control group, and both(BBT þ H þ L) and (H þ L) groups showed consistent nuclear immunoreactivity inthe epithelial cells but occasional and weaker intra-cytoplasmic positivity was

FIGURE 1 Photomicrographs of mammary tissues of rats with vehicle treatment (A, control),DMBA- treated rats (B) showed lobular hyperplasia where lobules have lumens, DMBA andCichorium treated rats (C) showed lobular hyperplasia, DMBA and light treated rats (D) showedhyperplasia with abounded stroma of fibrous tissue (Fibroadenoma), DMBA, Cichorium and lighttreated rats (E) showed normal lobules, and Cichorium þ light treated rats (F) showed normalmammary glands. Hematoxylin and Eosin staining, 400 £ .

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detected Fig. 2E. The remaining groups showed a significant increase of nuclear ERimmunoreactivity.

Effects on Lipid PeroxidationMDA values show different significance levels of elevation when compared withsham-control, whereas groups (BBT þ H), (BBT þ L), (H þ L) and (BBT þ W) havep , 0.001 and (BBT þ H þ L) has p , 0.01. The p-value of (BBT þ H) group vs.(BBT þ L) shows no significant variation. There is significant elevation inMDA levelsfor group (BBT þ H) compared with (BBT þ H þ L) (p , 0.05) as well as (BBT þ H)compared with (BBT þ W) (p , 0.01). No significant changes in MDA levels arefound in group (BBT þ H þ L) when compared with (BBT þ L) and (BBT þ W).

FIGURE 2 Immunohistochemical staining of ER in mammary tissues of rats with no treatment(A, control), benign breast cancer (B, induced), benign breast cancer þ Cichorium (C), benignbreast cancerþ L (D), benign breast cancerþ Cichoriumþ L(E), and Cichoriumþ L (F), Cichoriumtreatments decrease the number of ER-positive cells in tumor masses (D-F). Counter stained withhematoxylin. All images, 400 £ .

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Effects on Protein OxidationIn comparison with sham-control, groups (BBT þ H), (BBT þ L), and (BBT þ Hþ L)show no significant variations in hepatic PC. On the other hand, groups (H þ L) and(BBT þ W) showed significant elevation (p , 0.001). The p-value of (BBT þ H þ L)group vs. (BBT þ L), (BBT þ H), and (BBT þ W) showed no significant change.There is significant elevation in PC levels for group (BBT þ H) when compared withboth (BBT þ W) (p , 0.05) and (BBT þ L) (p , 0.01).

Effects on Antioxidants Defense SystemEffect of SODSOD levels were significantly decreased, in (BBT þ L, p , 0.001), (BBT þ W,p , 0.001), and (BBT þ H þ L, p , 0.01)) as compared to sham-control, while nosignificant change in (BBT þ H) and (H þ L). No significant changes were found incomparison of [(BBT þ H þ L) vs. (BBT þ L) and (BBT þ H); (BBT þ W) Vs(BBT þ L)]. A significant decrease was noticed in [(BBT þ W) vs. (BBT þ H,p , 0.05) and (BBT þ H þ L, p , 0.01).

Effect of CATThe antioxidant profiles in liver are shown in Table 1. The CAT levels are significantlydecreased, (p , 0.01) for both (BBT þ H) and (BBT þ H þ L), and (p , 0.001) for(BBT þ W) when compare to sham-control. Intra-comparison of other groups showno significant change in (BBT þ Hþ L) with (BBT þ L), (BBT þ H), and (BBT þ W).

Effect of TACTAC levels were significantly decreased (p , 0.001) in groups (BBT þ H) and(BBT þ W), and (BBT þ L) (p , 0.01), while no significant change in groups(BBT þ H þ L) and (H þ L) when compare to sham-control. No significant changein (BBT þ H þ L) vs. (BBT þ L) and (BBT þ H), while it is elevated significantlyfor (BBT þ W, p , 0.001). A significant decrease was noticed in group (BBT þ W) vs.(BBT þ L) (p , 0.05).

DISCUSSION

Interaction of photosensitizers (Chicory) with sunlight produces free radical thatreacts with oxygen and produce ROS. Other free radical reaction leads to theformation of highly reactive singlet oxygen (1O2). In biological tissues, either singletoxygen and/or ROS interact and cause cell death and tumor necrosis (Corner et al.,1996; Nevrelova et al., 2005). Chicory extracts was used experimentally to treatmalignant and benign diseases, also in clinical trials to treat tumors of the bronchus,bladder, esophagus, head and neck, brain, and skin (Gonier et al., 1989; Henderson,1989; Cerutti, 1994a).

ROS are normally produced in tissue cells. Over expression of their levels inbiological tissues causes oxidative stress which is measured using biomarkers asMDA and PC, whereas MDA produced from free radical attack to polyunsaturatedfatty acids and it is known to play an important rule in lipid peroxidation. PC contentis used as a biomarker for protein oxidation. To neutralize oxidative stress, cellsdeveloped antioxidant defense systems to protect lipid peroxidation, whichfunctioning in the level of scavengers and reflected by abnormal erythrocyteactivities of SOD, CAT, and TAC.

Our results show a positive significant elevation of bothMDA and PC, and negativesignificant difference of the antioxidative parameters when comparing the BBT withcontrol group. These significant differences are due to the reason that cancer cells

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TABLE1EffectofCichorium

(H)andligh

t(L)onhep

aticmarkersofoxidative

stress

(MDAandPC)andantioxidanten

zymes

(SOD,CAT,andTAC)in

controlandben

ign

breast

tumor(BBT)-groups.UnitsofandTAC,MDA,andPCare

nmol/mgprotein,unitsofSOD

andCATen

zymes

are

units/mgprotein.

MDA

PC

SOD

CAT

TAC

Mean^

SD

Mean^

SD

Mean^

SD

Mean^

SD

Mean^

SD

Groups

(p-value)

(p-value)

(p-value)

(p-value)

(p-value)

Sham

Control

0.193^

0.01

1.06^

0.09

13.68^

0.81

238.75^

03.85

54.29^

1.77

(Sham-C

ontrol)(W

)H

þL

0.224^

0.018

1.350^

0.137

12.788^

1.605

235.531^

11.881

53.448^

3.142

(0.001)

(0.001)

(0.234)

(0.959)

(0.615)

BBTþ

L0.227^

0.020

1.134^

0.167

9.900^

3.119

227.581^

14.741

49.099^

3.486

(0.001)

(0.371)

(0.001)

(0.065)

(0.003)

BBTþ

H0.241^

0.015

1.193^

0.191

11.438^

3.465

213.380^

14.985

48.334^

3.339

(0.001)

(0.268)

(0.105)

(0.003)

(0.001)

BBTþ

W0.216^

0.017

1.505^

0.189

8.300^

2.237

223.251^

7.597

45.491^

2.091

(0.001)

(0.001)

(0.001)

(0.001)

(0.001)

BBTþ

Hþ

L0.214^

0.024

1.275^

0.409

11.800^

2.108

224.099^

13.203

51.203^

5.091

(0.010)

(0.426)

(0.005)

(0.003)

(0.071)

(BBTþ

L)

––––

––––

––––

––––

––––

(0.153)

(0.713)

(0.161)

(0.798)

(0.214)

(Ben

ignBreast

Tumorþ

Herbþ

Light)

(BBTþ

H)

––––

––––

––––

––––

––––

(BBTþ

Hþ

L)

(0.035)

(0.916)

(0.505)

(0.130)

(0.092)

BBTþ

W––––

––––

––––

––––

––––

(0.711)

(0.092)

(0.010)

(0.234)

(0.001)

(Ben

ignBreast

Tumorþ

Water)

(BBTþ

L)

––––

––––

––––

––––

––––

BBTþ

W(0.316)

(0.004)

(0.161)

(0.798)

(0.036)

(BBTþ

H)

––––

––––

––––

––––

––––

(0.006)

(0.021)

(0.028)

(0.105)

(0.950)

(Ben

ignBreast

Tumorþ

Herb)

BBTþ

L––––

––––

––––

––––

––––

(BBTþ

H)

(0.291)

(0.431)

(0.798)

(0.083)

(0.648)

Resultsare

expressed

asmeans^

SD.Significantdifferencesbetweengroupswereco

nsidered

atp,

0.05.

Dashed

lines

representsamedata

forsamegroupmen

tioned

above.

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produce large amount of free radicals (Dormandy, 1983a,b; Cerutti, 1994b; Syed et al.,2004; Chan et al., 2009). The increase of lipid peroxidation causes higher levels ofMDA and PC which is associated with reduction in the levels of SOD, CAT, and TAC.These are consistent with results found by Chan et al. (Kisiel and Zielinska, 2001).Similar results also found when comparing H þ L with control group, whereas thereaction of the H and L produces ROS species. These findings support the previousresults that Cichorium contains some photosensitive compounds that could be usedas a potential drug for treating some types of benign tumors (Bischoff et al., 2004;Giese, 1980; Kisiel and Michalska, 2003; Ahmad et al., 2008).

MDA level significantly decreases for group administrated BBT þ H þ Lcomparing to those administrated with BBT þ H. This could be attributed tophotosensitizer activation that cause suppression to BBT cells activity and reducesthe oxidative stress. Even though both MDA and PC are oxidative stress markersand elevated compare to control, but the significant change in MDA levels arepoorly correlated with that of PC. This could be an indication that there is no PCvariation unless there is a severe change occurred (Yagci et al., 2008; Dalle-Donneet al., 2003; Amirkhizi et al., 2007). Further investigation of oxidative stress show thatrats administrated BBT þ H or BBT þ L show significant reduction on their PC level,and elevation in MDA with highly significant in BBT þ H. This could be due to theeffect of intra-reaction between sensitizer Cichorium, BBT, and ROS which is not veryclear to us. This result contradicts what was found by Petrovic et al. (Shindo et al.,1993), that the Cichorium is a strong antioxidant. To the best of our knowledge,no further similar studies or explanation are available in the moment to supportthis result.

This study reports the impact of the Cichorium as a photosensitizers agent onthe oxidative and antioxidative stress. All results show similar responds overall,whereas all show less values of SOD, CAT, and TAC compare to control. Thesereduction of antioxidative stress could be due to the elevation levels of oxidativestress. The effect of Cichorium as a photosensitizer on the antioxidative stress groupsadministrated BBT show reduction in their levels as compare to (BBT þ H þ L),which could be due to the hypothesis that the sensitizer induces apoptosis in cancercells and tumor cells. This result does not valid for CAT which shows poor correlationwith TAC (Vertechy et al., 1989; [49]). SOD is considered to be the first line in thedefense system that converts the superoxide anion (O2

1 or ·2) into hydrogen peroxide(H2O2) which in turns removed by catalase. So CAT could be less dependent andas a second line in the defense system. Also various antioxidant markers reactdifferently and poorly correlate (Vertechy et al., 1989; [49]).

The influence of Cichorium as a photosensitizer agent appears on the ERdistributions (Fig. 2), whereas samples from control group, group (H þ L), andgroup (BBT þ H þ L) showed consistent nuclear immunoreactivity in the epithelialcells compare to the cells with BBT. The Cichorium treatments significantly decreasethe number of ER-positive cells in tumor masses.

CONCLUSION

In the present work, the concomitant treated with Cichorium and light reduced thelobular hyperplasia and fibroadenoma induced by DMBA in female mammaryglands. It also found that Cichorium and light treatments decreased the number ofER-positive cells in mammary glands. These effects were accompanied withincreased in oxidative stress markers (MDA, SOD, CAT, and TAC) in mammarytissues. This indicated that the influence of the Cichorium and light againstpathological effect of DMBA on mammary glands could be attributed to free

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radical production. The results of the present study clearly demonstrated thatCichorium act as photosensitizing agent against breast cancer by inducing oxidativestress damage.

ACKNOWLEDGEMENTS

This work was funded by the scientific research council of United Arab Emirates,grant # 08-02-2-11/06, and partially by higher council for science and technology/Jordan, grant # 62/2000.The authors are grateful toMiss. Fedae Alhaddad,Miss. NadaAl-Kahlout, and to Mr. Taleb for their valuable assistance throughout this work.

Declaration of interest

The authors report no conflicts of interest. The authors alone are responsible for thecontent and writing of this article.

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