EFFECT OF CIPROFLOXACIN, VITAMIN E AND THEIR …

www.wjpps.com Vol 7, Issue 3, 2018.

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El-Sayed et al. World Journal of Pharmacy and Pharmaceutical Sciences

EFFECT OF CIPROFLOXACIN, VITAMIN E AND THEIR

COMBINATION ON DNA FRAGMENTATION AND BONE MARROW

CYTOGENICITY

El-Sayed M. G. A.*1, A. A. A. El-Komy

1, Hanan M. Sobhy

2 and Shimaa G. Saied

2

1Department of Pharmacology, Faculty of Veterinary Medicine, Benha Univerisity.

2Department of Biochemistry, Animal Health Research Institute, Dokki, Giza.

ABSTRACT

The antibacterial activities of the fluorinated 4-quinolone,

ciprofloxacin has been ascribed to a marked inhibition of bacterial

DNA gyrase. However, evidence is accumulating that ciprofloxacin

may cause liver DNA damage. Vitamin E is a free radical scavenger

and antioxidant. This study aimed to investigate the influence of

ciprofloxacin in recommended dose on liver and spleen deoxy-

ribonucleic acid as well as on micronuclei formation in bone marrow

erythrocytes and the effects of vitamin E on these changes.Sixty adult

male Sprague Dawley rats were divided into four equal groups:

control, ciprofloxacin treated, vitamin E treated and ciprofloxacin and

vitamin E treated. DNA fragmentation was evidenced in liver and

spleen cells of ciprofloxacin treated rats.Micronuclei formation significantly increased in

ciprofloxacin treated rats compared to control group. Vitamin E administration reduced liver

DNA fragmentation caused by ciprofloxacin in rats. It is suggested that.

KEYWORDS: Ciprofloxacin, Vitamin E, DNA, Cytogenicity.

INTRODUCTION

Ciprofloxacin is a flourinated quinolone carboxylic acid derivative which was developed for

use exclusively in animals. It exhibits a wide spectrum of antimicrobial activity, like other

fluoroquinolones including some Gram positive, Gram negative bacteria and Mycoplasma.[1]

Ciprofloxacin acts by the inhibition of the A subunit of the bacterial DNA gyrase which is an

essential enzyme for DNA replication and synthesis. This synthetic drug acts as bactericidal

Article Received on

21 Jan. 2018,

Revised on 11 Feb. 2018,

Accepted on 02 March 2018

DOI: 10.20959/wjpps20183-11080

*Corresponding Author

El-Sayed M. G. A.

Department of

Pharmacology, Faculty of

Veterinary Medicine, Benha

Univerisity.

[email protected]

WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

SJIF Impact Factor 7.421

Volume 7, Issue 3, 1045-1060 Research Article ISSN 2278 – 4357

mailto:[email protected]


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at relatively low concentration[2]

The type II topoisomerases DNA gyrase and topoisomerase

IV is the target of quinolones.[3]

Vitamin E is the major lipid soluble antioxidant responsible for protecting membranes against

lipid peroxidation and is the term for a group of tocopherals and tocotrienols, of which -

tocopherots, has the highest biological activity.[4]

Most alpha-tocopheral is located in the

mitochondrial fractions and in the endoplasmic reticulum of the cells.[5]

The possible protective effect of vitamin E to the genotoxicity induced by fipronil, a

phenylpyrazol insecticide classified as a carcinogen by United States Environmental

Protection Agency was studied. The structural chromosome aberrations in bone marrow cells

and deoxy-ribonucleic acid damage in the lymphocytes was found to be significantly higher

in the male and female rats exposed to fipronil as compared to their respective controls. The

average degree of protection (male and female animals combined together) shown by

pretreatment of vitamin E against fipronil-induced genotoxicity was 63.28%: chromosome

aberrations; 47.91%: micronuclei formation; and 74.70%: deoxy-ribonucleic acid damage.[6]

The aim of the present work was undertaken to study the cytotoxic effect of ciprofloxacin on

cellular deoxy-ribonucleic acid in vivo and to declare the role of vitamin E as an antioxidant

to protect cellular deoxy-ribonucleic acid. Also to study Effect of ciprofloxacin, vitamin E

and their combination on bone marrow cytogenicity.

MATERIALS AND METHODS

Materials

1-Drugs

Ciprofloxacin is a flouroquinolone available as white to yellowish white powder under trade

name CIPROVAN®

20% (Turkey)-it is easily soluble in water. The drug used directly after

reconstitution in water for oral administration. Vitamin E is available as oral gelatinous

capsules contain yellowish oily substance. It was purchased from PHARCO Company,

Egypt. It is chemically α-Tochopherol, 5,7,8- trimethyltocol. Ciprofloxacin and vitamin E

were administrated in a dose of 12.5mg/ Kg b.wt and 100mg/kg b.wt by stomach tube once

daily for one week.[7,8]

Agrose gel (1%), TAE buffer (0.5x Tris Acetate EDTA buffer),

ethydium/bromide and Marker (M) 100-bp ladder size marker were obtained from Sigma

company. Kits for DNA extraction obtained from Intron Company, Korea.


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2-Rats

Sixty mature male rats of an average body weight 150-180 grams of Sprague Dawley strain

were obtained from Animal Health Research Institute Giza, Egypt. Basal diet supplying the

essential vitamins and trace elements was prepared.[9]

METHODS

Experimental design

The experiment was done in the pharmacology unit, Biochemistry, Toxicology and Food

Deficiency Department, Animal Health Research Institute. Animals were acclimatized to

laboratory conditions for one week. The animals were kept under hygienic conditions, housed

in metal cages containing wood shaving as bedding material. Rats kept under controlled

temperature at 23°C ± 2°C and maintained on a 12/12 h light and dark cycle during the

experiment with nonstop access to rat chow and tap water. The use of animals in the study

was in accordance to the guidelines of National Research Council for the care and use of

Laboratory Animals. Rats fed on basal diet and water supply was given ad-libtium all over

the period of the experiment. Rats were divided into four groups which were group (1) that

act as negative control, group (2) that given ciprofloxacin, group (3) that given vitamin E and

group (4) that given vitamin E as pretreatment followed by ciprofloxacin for one week.Liver

and spleen from each sacrificed rat were collected in sterilized eppindrof tubes and kept in

deep freeze (-80oC) for DNA fragmentation study. Also, bone marrow samples extruded from

femur of control and treated rats were collected separately and fixed immediately with

ethanol for cytogenic assay.

DNA fragmentation [DNA ladder Assay]: DNA Damage Inhibition Analysis was done in

Genetic Research Center El-Sadat University. The extent of DNA fragmentation (DNA

ladder) has been assayed by electrophoresing genomic DNA samples, isolated from normal

as well as treated rat livers and spleens; on agrose/ethydium bromide gel.[10]

High quality

genomic DNA was extracted from (-80oC preserved liver and spleen samples of all treated

and control groups) by precipitation of protein and other contents and further precipitation of

high molecular weight genomic DNA by absolute ethanol[11]

The amplified products of

different exons were verified in 1% (W/V) agrose gel. DNA molecules are separated by

applying an electric field to move the negatively charged molecules through an agrose matrix.

Shorter molecules move faster and migrate farther than longer ones because shorter

molecules migrate more easily through the pores of the gel.


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Bone marrow cytogenetic assay: In order to investigate the ameliorating effect of vitamin E

on the possible mutagenic effects of ciprofloxacin, the micronucleus test was performed to

detect chromosomal damage associated with the treatment. Micromuclei were identified as

dark blue staining bodies in the cytoplasm of the poly chromatic erythrocytes (PCEs). Using

standard laboratory technique[12]

,bone marrow cells of control and treated rats were extruded

with a pin into a clean dry glass slide and homogenized with two drops of fetal calf serum on

the slide, air dried, fixed in absolute methanol and stained with Giemsa 5% in phosphate

buffer pH 6.8.The pollychromatic erythrocytes (PCEs, 1000 per animal) were screened for

micronuclei, and the changes in the mitotic activity[13,14]

were assessed on the basis of the

ratio of polychromatic to normochromatic erythrocytes (PCE/NCE ratio).

STATISTICAL ANALYSIS

Statistical analysis was performed using one-way analysis of variance (ANOVA) for multiple

comparisons of all groups. Means and there standard errors (SE) were calculated using the

SPSS 16 software, followed by Least Significant Difference test (LSD) to calculate the

statistical difference between various groups. A value of P<0.05 was considered statistically

significant.[15]

RESULTS

Effect of ciprofloxacin, vitamin E and its combination on DNA fragmentation

Liver DNA fragmentation on agrose/ethydium bromide gel, DNA isolated was loaded onto

1% (W/V) agrose gels compared to Marker (M) 100-bp ladder size marker at one day post

last treatment as in figure (1). Lane (1) represented DNA isolated from liver tissues of

negative control rats revealed a single band corresponding to intact nuclear DNA. Lane (2)

represented DNA isolated from ciprofloxacin treated liver samples revealed smear formation

that indicated DNA damage and necrotic cell death to hepatic cells. Lane (3) represented

DNA extracted from liver samples treated with vitamin E revealed intact DNA as one nuclear

band. Lane (4) represented DNA extracted from liver samples of rats treated with both

ciprofloxacin and vitamin E at the same time which revealed incomplete protective action of

vitamin E to nuclear DNA fragmentation.


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Figure. (1): DNA isolated from experimental liver tissues 1 day post last treatment on

agarose/ethydium bromide gel.

Liver DNA extracted from liver samples at 7 days post last treatment. The protective effect of

vitamin E to the necrotic cell death is clearly visible in lane (4), as the gels revealed no smear

formation while only one migrating band of low molecular weight in each lane in comparison

with smear and band formation in lane (2) that represent DNA extracted from liver cells of

ciprofloxacin treated rats as represented in figure (2).

Figure. (2): DNA isolated from experimental liver tissues 7 days after stopping of

treatment on agarose/ethydium bromide gel.

DNA extracted from liver samples at 14 days post last treatment showed that both lane (2)

and lane (4) revealed weak staining quality of extracted genomic DNA from liver samples

suggesting DNA diminution in hepatocytes. Lane (4) showed no smear formation in

comparison with lane (2) because of a consequence of low DNA degradation as represented

in figure (3).


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Figure. (3): DNA fragmentation on agarose/ethydium bromide gel. DNA isolated from

experimental liver tissues 14 days after stop of treatment.

DNA fragmentation on agarose/ethydium bromide gel isolated from spleen of rats at one day

after stop of treatment was represented in figure (4). Lane (1): represents DNA extracted and

electrophoresed from spleen of negative control samples revealed a single nuclear band. Lane

(2) represents DNA extracted from spleen samples of ciprofloxacin treated rats exhibited

multiple fragments evidenced by a ladder – like separation of DNA fragments on the gel.

Lane (3) represented DNA extracted from spleen samples of vitamin E treated rats, exhibited

intact nuclear DNA. Lane (4) represents DNA extracted from spleen samples ciprofloxacin

and vitamin E treated rats revealed that vitamin E is effective to some extent in protection of

the cells and DNA fragmentation.

Figure. (4): DNA isolated from experimental spleen tissues 1 day after stop of treatment

agrose / ethydium bromide gel.


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DNA fragmentation on agarose/ethydium bromide gels, isolated from spleen samples of rats

at 7 days after stop of treatment. Lane (2) showed ladder like separation of genomic DNA

fragments to some extent due to stopping of treatment on week later, while in lane (4), no

ladder like fragmentation as found in figure (5).

Figure. (5): DNA isolated from experimental spleen tissues 7 days after stopping of

treatment on agarose/ethydium bromide gel.

DNA extracted from spleen samples at 14 days post last treatment was represented in figure

(6). Lane (2) represents DNA extracted from spleen samples of ciprofloxacin treated rats. It

revealed smear formation but no clear ladder – like separation of genomic DNA as a result of

decrease oxidative stress of ciprofloxacin on spleen cells by time. Lane (4) revealed intact

migrating bands and slight smear formation, due to antioxidant role of vitamin E on DNA

damage induced by ciprofloxacin.

Figure. (6): DNA fragmentation on agarose/ethydium bromide gel. DNA isolated from

experimental spleen tissues 14 days after stop of treatment.


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Effect of ciprofloxacin, vitamin E and its combination on bone marrow cytogenicity

The obtained results showed that treatment with ciprofloxacin produced significant decrease in normochromatic erythrocytes (NCE) at one and

seven days post last treatment and increase in multinucleated polychromatic erythrocytes (MPCE) as well as the ratio of polychromatic

erythrocytes to normochromatic erythrocytes (PCE/NCE) compared to control group. Vitamin E treatment produced no change in

normochromatic erythrocyts (NCE) 1, 7and 14 days post last treatment compared with control group. Rats treated with both ciprofloxacin and

vitamin E (group 4) indicated significant increase in normochromatic erythrocytes (NCE) compared to ciprofloxacin treated group 1and 7 days

post last treatment as well as decrease in multinucleated polychromatic erythrocytes(MPCE) and the ratio of polychromatic erythrocytes to

normochromatic erythrocytes(PCE/NCE) compared to control group (group1), 1and 7 days post last treatment as illustrated in table 1 and figure

7,8 and 9.

Table. (1): Effect

of ciprofloxacin, vitamin E and their combination on the incidence of multinucleated polychromatic

erythrocytes(MPCE), normochromatic erythrocytes(NCE) and polychromatic erythrocytes / normochromatic erythrocytes

ratio(PCE/NCE), (n=5).

Groups PCE

NCE MPCE PCE/NCE

Time post treatment Time post treatment Time post treatment

1 day 7days 14days 1 day 7days 14days 1 day 7days 14days

Group (1) 5000 2156.67±

9.63a

2159.4±

12.46a

2148.98±

23.82a

5.32±

0.20a

5.25±

0.15a

5.15±

0.13a

2.22±

0.30a

2.39±

0.17a

2.55±

0.11a

Group (2) 5000 740.39±

11.49 b

824.82±

29.77b

2130.21±

5.00b

14.58±

0.43 b

9.22±

0.072b

5.63±

0.11b

6.77±

0.22 b

5.81±

0.24 b

2.83±

0.26b

Group (3) 5000 2153.86±

20.19c

2183.9±

31.58c

2156.92±

5.59c

5.86±

0.12c

5.67±

0.27c

5.26±

0.14c

2.10±

0.10a

2.32±

0.09a

2.03±

0.14a

Group (4) 5000 944.63±

32.18 b

961.79±

18.02 b

2149.81±

41.09b

9.14±

0.24 d

7.04±

0.13 d

5.75±

0.15d

3.78±

0.12 c

3.70±

0.10 c

2.76±

0.18b


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Figure. (7): Effect

of ciprofloxacin, vitamin E and their combination on

Normochromatic erythrocytes (NCE).

Figure (8): Effect

of ciprofloxacin, vitamin E and their combination on multinucleated

polychromatic erythrocytes.

Figure (9): Effect

of ciprofloxacin, vitamin E and their combination on polychromatic

erythrocytes/ Normochromatic erythrocytes.

G(1):Untreated control rats.

G((2):Rats treated with ciprofloxacin(12.5 mg/kg b.wt) for 7 days.

G(3):Rats treated with vitamin E (100mg/kg b.wt.) for 7 days.

G(4):Rats treated with ciprofloxacin (12.5 mg/kg b.wt) and vitamin E(100mg/kg b.wt.) for 7

days.


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DISCUSSION

It is known that ciprofloxacin could inhibit of gyrase, a specific prokaryot enzyme that

catalyses the conversion of relaxed DNA into negatively supercoiled DNA. Acombination of

gyrase and topoisomerase-I is required to correct DNA topology during replication and

transcription. The inhibition of gyrase by quinolones triggers replication arrest and cell

death.[16]

The obtained results showed that the cellular DNA isolated from splenic and hepatic

cells, one day post last treatment in ciprofloxacin treated group evidenced clear fragmentation

and distinct ladder-like appearance especially in splenic DNA. Explaining the results,

Bredberg et al., (1991)[17]

reported that ciprofloxacin-induced marked induction of double-

strand DNA breaks in human lymphoblastoid cells. Moreover, Albertini et al., (1995)[18]

declared that, ciprofloxacin inhibit resolution of the normally transient topoisomerase II-

DNA cleavage complex resulting in chromosome stickness. This was supported later by

Gürbay et al., (2007)[19]

, who reported that ciprofloxacin generated reactive oxygen species

(Hydrogen peroxide) which considered one of most free radical causing cellular protein

damage and apoptosis in primary cultures of rat astrocytes and normal human fibroblast cells.

On the other hand, the obtained results of DNA fragmentation in ciprofloxacin treated rats

seven days post last treatment showed DNA damage in hepatic cells, suggesting the

continuation of impact action of ciprofloxacin through generation of reactive oxygen species

resulted in liver cellular damage.[20,21]

Results of splenic DNA fragmentation, seven days

following last treatment, are in harmony with that obtained by Klaudia and Alina, (2015)[22]

who reported that enrofloxacin, florfenical and ceftiofur administrated in therapeutic doses

may decrease the percentage of B and T cells subset in lymphatic organs and might influence

on the proportion of B and T cells in chicken spleen and thymus. The results of DNA

fragmentation are parallel fomer studies.[23-26]

The obtained results, 14 days post treatment for

hepatic and splenic genomic DNA indicated decreased smear formation isolated form

ciprofloxacin treated rats and low DNA degradation, this could be explained by that the

ciprofloxacin concentration, decrease in the cellular fluid by time, so it is evidenced from the

study that the effect of ciprofloxacin on DNA might be reversible. In contrary with our results

at one and 7 days post treatment, Herbold et al., (2001)[27]

mentioned that ciprofloxacin tested

for in vivo genotoxicity revealed no genotoxic effect using the following test systems:

micronucleus test in bone marrow of mice, cytogenetic chromosome analysis, in Chinese

hamster, dominant lethal assay in male mice and UDS tests in primary rat and mouse

hepatocytes in vivo. In addition, ciprofloxacin not found to be carcinogenic in two rodent


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long-term bioassays. On the other hand Smart et al., (2008)[28]

suggested that DNA-douple

strand breaks are not induced by 0-100 g/ml ciprofloxacin (0-10 hours exposure) in human

derived cells, therefore ciprofloxacin –induced clastogenicity in such cells arised from DNA

damage is different to direct DNA double strand breaks, moreover, the study didn't rule out

the possibility of other types of ciprofloxacin-mediated DNA damage as single strand DNA

lesions.

Regarding DNA fragmentation in hepatic cells, in ciprofloxacin and vitamin E group(G1),

one and seven days post treatment, Lane (4) in figure (1) and (2) revealed partial protection

by vitamin E, as decreased, smear formation with migration of low molecular weight nucleic

acid bands. Furthermore, vitamin E has no protective action on splenic DNA degradation by

ciprofloxacin in group(4),one day post treatment as evidenced by ladder-like separation of

splenic DNA, showed in lane (4), figure (4). One the other hand, vitamin E, showed a clear

protection in lane (4) in both figure (5) and (6) that representing DNA isolated from splenic

cells of ciprofloxacin treated rats, seven and 14 days respectively following last treatment, as

that genomic DNA has a likely an intact appearance with high molecular weight and low

degradation model.

Explaining the obtained results, about role of vitamin E in monitoring cytotoxic effect of

ciprofloxacin on hepatic and splenic cells, we can say that, vitamin E is mainly a chain-

breaking antioxidant against membranal damage, but probably not as effective against the

damage at mitochondrial level[19],

so vitamin E couldn't reverse the induction of oxidative

DNA damage by ciprofloxacin in hepatic and splenic cells as evidenced by the obtained

results one day post treatment. Moreover, vitamin E should be used as a pretreatment to this

drug, this supported by Mozdarani and Salimi, (2006)[29]

who declared that administration of

vitamin E before irradiation effectively reduced the frequency of chromosomal abnormalities.

The partial protective effect of vitamin E, seven and 14 days post treatment, declared its role

as radical scavenger. Similar results of vitamin E were also observed.[6,30-33]

According to the results of ciprofloxacin effect on micronuclei formation in bone marrow

erythrocytes, The administration of ciprofloxacin at therapeutic dose for 7 days causing

increase in multinucleated polychromatic erythrocytes statistically following stopping of

administration by one and seven days in comparison with control group. The micronecleus

assay is devised primarily for evaluating the ability of test agent to induce structural and/or


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numerical chromosomal damages. This results come in accordance with Curry et

al.,(1996)[23]

who found that structural chromosomal aberrations detected in metaphase cells

was attributed to ciprofloxacin-inhibition of topoisomerase II prior to mitosis while

micronuclei arised in binucleated cells as a result of this effect which interfers with

chromosomal separation. In the same manner, Olajuyigbe et al.,(2011)[34]

declared that

apossible mechanism for the induction of micronuclei in the mouse bone marrow cells is that

ciprofloxacin inhibited topoisomerase II in mice exposed to different concentrations of

ciprofloxacin (16,32,80,160, 320 µg/ml). In-vitro study by Ṥekeroğlu et al.,(2017).[26]

investigated the cytotoxic and genotoxic potentials of ofloxacin in cultured human peripheral

lymphocytes. Cultures were treated with 30, 60 and 120 μg/ml of ofloxacin for 48 hours.

Ofloxacin significantly induced chromosomal aberrations at all concentrations and sister

chromatid exchanges at higher concentrations (60 and 120 μg/ml) compared with solvent

control. El-Habit, Ola, et al (2001)[35]

suggested that the direct assessment of genotoxicity of

ofloxacin was achieved by scoring the incidence of multinucleated polychromatic

erythrocytes and multinucleated normochromatic erythrocytes. The results demonstrated a

positive dose response relationship, which suggested that ofloxacin is mutagenic at high

doses. The equivalent therapeutic dose, however, did not induce marked increase in the

incidence of micronucleated erythrocytes. The discrepancies noted between different reports

can be attributed to differences in dose, rout of administration, duration of treatment or test

system. In contrary with the present study, Herbold et al., (2001)[27]

mentioned that

ciprofloxacin tested for in vivo genotoxicity revealed no significant increase in the number of

micronucleated erythrocytes in bone marrow of mice.

However,vitamin E in combination with ciprofloxacin decreased the level of micronuclei

formation and polychromatic erythrocytes to normochromatic erythrocytes ratio. This

supported by Chorvatovičová.,et al., (1991)[36]

who found that the micronucleus test in bone

marrow showed that vitamin C caused antimutagenic effect against bichromate, in both rats

and guinea pigs,while the effect of vitamin E was demonstrated only in an increase of the

ratio of normochromatic erythrocytes to polychromatic erythrocytes, i.e., in a decrease of the

cytotoxic but not the mutagenic effects of hexavalent chromium. Moreover, Konopacka and

Rzeszowska-Wolny (2001)[37]

decleared that in-vitro study, human lymphocytes treated with

vitamin E at a concentration above 2 mg/ml showed a decrease at the level of radiation-

induced micronuclei when compared to the cells irradiated without vitamin treatment. Singh

et al., (2008)[8]

demonstrated that vitamin E along with atrazine, decrease the percentage of

http://www.sciencedirect.com/science/article/pii/016579929190104C#!


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micronuclei as compared to atrazine treated rats. Furthermore, Badgujar et al.,(2017)[6]

suggested that adult male and female animals gavaged with various doses of fipronil (2.5,

12.5, and 25 mg/kg body weight to evaluate micronucleus test (mice), chromosome

aberration, and comet assay (rats), respectively and were pretreated with vitamin E orally

(400 mg/kg body weight) for 5 days prior to administration of fipronil (12.5 mg/kg).

Pretreatment of vitamin E induced a decrease in fipronil-induced micronuclei formation by

47.91%.

CONCLUSION

The results of the present study suggested that ciprofloxacin induce genotoxicity on basis of

cellular deoxy-ribonucleic acid. Moreover,micronuclei formation and increase in immature

erythrocytes were evidenced. Treatment with a known antioxidant, vitamin E has shown

protective effect to DNA fragmentation.

REFERENCES

1. Oliphant, C. M., & Green, G. M., Quinolones: a comprehensive review. American family

physician, 2002; 65(3): 455-64.

2. Brown, S.A.,Fluoroquinolones in animal health. J. Vet. Pharmacol. Therap., 1996; 19:

1-14.

3. Champoux, J. J.,0 DNA topoisomerases: structure, function, and mechanism. Ann. review

of biochem, 2001; 70(1): 369-413.

4. Brigelius-Flohe, R., & Traber, M. G.,Vitamin E: function and metabolism. The FASEB

J., 1999; 13(10): 1145-55.

5. Drevon, C. A.,Absorption, transport and metabolism of vitamin E. Free radical research

communications, 1991; 14(4): 229-46.

6. Badgujar PC, Selkar NA, Chandratre GA, Pawar NN, Dighe VD, Bhagat ST, Telang AG,

Vanage GR. Fipronil-induced genotoxicity and DNA damage in vivo: Protective effect of

vitamin E. Human & experimental toxicology, 2017; 36(5): 508-19.

7. Abu-Aita, N.A.; Ahmed, K.A. and Mouneir, S.M. The protective effect of ginger and

Nacetyl cysteine on ciprofloxacin-induced reproductive toxicity in male rats. Journal of

American Science, 2011; 7(7): 741-52.

8. Singh, M., Kaur, P., Sandhir, R., &, Kiran, R., Protective effects of vitamin E against

atrazine-induced genotoxicity in rats., Mutation Research/Genetic Toxicology and

Environmental Mutagenesis, 2008; 654(2): 145-49.‏

http://het.sagepub.com/search?author1=PC+Badgujar&sortspec=date&submit=Submit


1058


9. NRC "National Research Council": Nutrient requirement of laboratory. Fourth reviser

edition. Pp: 29-30 National Academy Press Washington, animals, D.C. Environ. Sci.

Health, 1995; 25: 487-494.

10. Sellins, K. S., & Cohen, J. J., Gene induction by gamma-irradiation leads to DNA

fragmentation in lymphocytes., The Journal of Immunology, 1987; 139(10): 3199-206.‏

11. Sambrook J, Fritsch EF and Maniatis T.,Molecular cloning: A laboratory manual. 2nd

edition, Cold Spring Harbor Laboratory, CSH, NY:1989.

12. Salamon, M.;J. Heddle; E. Suart & M. Katz,Towards an improved Studies on 3 model.

Mitomycin C, cyclmicronucleus test:counting, distribution and ophosphamide and

dimethyl benzanthracene., Mutat. Res., 1980; 74: 347-56.

13. Hart JW, Engberg-Pedersen H. Statistics of the mouse bone-marrow micronucleus test:

counting, distribution and evaluation of results, Mutation Research/Fundamental and

Molecular Mechanisms of Mutagenesis. 1983; 111(2):195-207.

14. Al-Bekairi AM, Qureshi S, Chaudhry MA, Shah AH. Uric acid as an inhibitor of

cyclophosphamide-induced micronuclei in mice. Mutation Research Letters, 1991;

262(2): 115-8.

15. Artimage, G.Y. & W.G. Berry. Statistical Methods 7th Ed. Ames, Iowa Stata University

Press, 1987; 39-63.

16. Drlica, K., & Franco, R. J.,Inhibitors of DNA topoisomerases. Biochem., 1988; 27(7):

2253-2259.

17. Bredberg, A. N. D. E. R. S., Brant, M. A. R. T. A., & Jaszyk, M. A. L. G. O. R. Z. A. T.

A.. Ciprofloxacin-induced inhibition of topoisomerase II in human lymphoblastoid

cells. Antimicrobial agents and chemotherapy, 1991; 35(3): 448-50.

18. Albertini S, Chételat AA, Miller B, Muster W, Pujadas E, Strobel R, Gocke E.

Genotoxicity of 17 gyrase-and four mammalian topoisomerase II-poisons in prokaryotic

and eukaryotic test systems. Mutagenesis, 1995; 10(4): 343-51.

19. Gürbay, A., Gonthier, B., Barret, L., Favier, A., & Hıncal, F., Cytotoxic effect of

ciprofloxacin in primary culture of rat astrocytes and protection by vitamin E.

Toxicology, 2007; 229(1): 54-61.‏

20. Dharnidharka, V. R., Nadeau, K., Cannon, C. L., Harris, H. W., & Rosen, S.,

Ciprofloxacin overdose: acute renal failure with prominent apoptotic changes. Am. J. of

Kid. Dis., 1998; 31(4): 710-12.

21. Pouzaud, F., Dutot, M., Martin, C., Debray, M., Warnet, J. M., & Rat, P., Age-dependent

effects on redox status, oxidative stress, mitochondrial activity and toxicity induced by


1059


fluoroquinolones on primary cultures of rabbit tendon cells., Comparative Biochemistry

and Physiology Part C: Toxicology & Pharmacology, 2006; 143(2): 232-41.

22. Klaudia, C., & Alina, W., The influence of enrofloxacin, florfenicol, ceftiofur and E. coli

LPS interaction on T and B cells subset in chicks. Veterinary research

communications, 2015; 39(1): 53-60.

23. Curry, P. T., Kropko, M. L., Garvin, J. R., Fiedler, R. D., & Theiss, J. C.,In vitro

induction of micronuclei and chromosome aberrations by quinolones: possible

mechanisms. Mutation Research/Fundamental and Molecular Mechanisms of

Mutagenesis, 1996; 352(1): 143-50.

24. Lawrence, J. W., Darkin‐Rattray, S., Xie, F., Neims, A. H., & Rowe, T. C., 4‐Quinolones

cause a selective loss of mitochondrial DNA from mouse L1210 leukemia cells.Journal of

cellular biochemistry, 1993; 51(2): 165-74.

25. Poulsen, K. L., Olivero-Verbel, J., Beggs, K. M., Ganey, P. E., & Roth, R. A.,

Trovafloxacin Enhances Lipopolysaccharide-Stimulated Production of Tumor Necrosis

Factor-α by Macrophages: Role of the DNA Damage Response. Journal of Pharmacology

and Experimental Therapeutics, 2014; 350(1): 164-70.‏

26. Şekeroğlu, V., Aksoy, M., & Atlı Şekeroğlu, Z.,Cytogenetic alterations in human

lymphocyte cultures following exposure to ofloxacin., Drug and chemical toxicology,

2017; 40(2): 140-45.

27. Herbold, B. A., Brendler-Schwaab, S. Y., & Ahr, H. J.,Ciprofloxacin: in vivo

genotoxicity studies. Mutation Research/Genetic Toxicology and Environmental

Mutagenesis, 2001; 498(1): 193-205.‏

28. Smart, D. J., Halicka, H. D., Traganos, F., Darzynkiewicz, Z., & Williams, G. M.,

Ciprofloxacin-induced G2 arrest and apoptosis in TK6 lymphoblastoid cells is not

dependent on DNA double-strand break formation, Cancer biology & therapy, 2008; 7(1):

‏.113-19

29. Mozdarani, H., & Salimi, M., Numerical chromosome abnormalities in 8-cell embryos

generated from γ-irradiated male mice in the absence and presence of vitamin E.

International journal of radiation biology, 2006; 82(11): 817-22.‏

30. Kaymak, C., Kadioglu, E., Basar, H., & Sardas, S.. Genoprotective role of vitamin E and

selenium in rabbits anaesthetized with sevoflurane. Human & experimental toxicology,

‏.413-19 :(8)23 ;2004

31. Porrini, M., Riso, P., Brusamolino, A., Berti, C., Guarnieri, S., & Visioli, F., Daily intake

of a formulated tomato drink affects carotenoid plasma and lymphocyte concentrations


1060


and improves cellular antioxidant protection. British Journal of Nutrition, 2005; 93(01):

‏.93-9

32. Laurent, C., Voisin, P., & Pouget, J. P., DNA damage in cultured skin microvascular

endothelial cells exposed to gamma rays and treated by the combination pentoxifylline

and α-tocopherol. International journal of radiation biology, 2006; 82(5): 309-21.‏

33. Rezaei-Basiri, M., Rezazadeh, H., Aswadi-Kermani, I., & Ghazi-Khansari, M., Anti-

mutagenicity Effects of Vitamin E on Oncology and Non-oncology Hospital Nurses by

Ames Assay., Journal of clinical and diagnostic research: JCDR, 2013; 7(12): 2917.‏

34. Olajuyigbe, O., Alabi, O., & Anaba, U., Induction of micronucleus and abnormal sperm

cells in albino mice exposed to ciprofloxacin.Pharm. Lett, 2011; 3(2): 438-45.

35. Ola, El-Habit, H., Abd-Allah, A. R., & El-Sais, S. N., Comparison of genotoxicity of

ofloxacin in normal and immune stressed mice. Suez Canal Univ. Med., 2001; 4: 1-8.

36. Chorvatovičová, D., Ginter, E., Košinová, A., & Zloch, Z.,Effect of vitamins C and E on

toxicity and mutagenicity of hexavalent chromium in rat and guinea pig. Mutation

Research Letters, 1991; 262(1): 41-46.

37. Konopacka, M., & Rzeszowska-Wolny, J., Antioxidant vitamins C, E and β-carotene

reduce DNA damage before as well as after γ-ray irradiation of human lymphocytes in

vitro. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 2001;

‏.1-7 :(1)491

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