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Influence of Ginger Rhizome (Zingiber officinale Rosc) on Survival, Glutathioneand Lipid Peroxidation in Mice after Whole-Body Exposure to Gamma RadiationAuthor(s): Ganesh Chandra Jagetia, Manjeshwar Shrinath Baliga, Ponemone Venkatesh, and Jagadish N.UlloorSource: Radiation Research, 160(5):584-592. 2003.Published By: Radiation Research SocietyDOI: http://dx.doi.org/10.1667/RR3057URL: http://www.bioone.org/doi/full/10.1667/RR3057

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RADIATION RESEARCH 160, 584–592 (2003)0033-7587/03 $15.00q 2003 by Radiation Research Society.All rights of reproduction in any form reserved.

Influence of Ginger Rhizome (Zingiber officinale Rosc) on Survival,Glutathione and Lipid Peroxidation in Mice after Whole-Body Exposure

to Gamma Radiation

Ganesh Chandra Jagetia,a,1 Manjeshwar Shrinath Baliga,a Ponemone Venkatesha and Jagadish N. Ulloorb

a Department of Radiobiology, Kasturba Medical College, Manipal 576 119, India; and b Department of Psychiatry and Program in BehavioralNeuroscience, Boston University School of Medicine, Boston, Massachusetts

Jagetia, G. C., Baliga, M. S., Venkatesh, P. and Ulloor, J.N. Influence of Ginger Rhizome (Zingiber officinale Rosc) onSurvival, Glutathione and Lipid Peroxidation in Mice afterWhole-Body Exposure to Gamma Radiation. Radiat. Res. 160,584–592 (2003).

The radioprotective effect of the hydroalcoholic extract ofginger rhizome, Zingiber officinale (ZOE), was studied. Micewere given 10 mg/kg ZOE intraperitoneally once daily for fiveconsecutive days before exposure to 6–12 Gy of g radiationand were monitored daily up to 30 days postirradiation forthe development of symptoms of radiation sickness and mor-tality. Pretreatment of mice with ZOE reduced the severity ofradiation sickness and the mortality at all doses. The ZOEtreatment protected mice from GI syndrome as well as bonemarrow syndrome. The dose reduction factor for ZOE wasfound to be 1.15. The optimum protective dose of 10 mg/kgZOE was 1⁄50 of the LD50 (500 mg/kg). Irradiation of the ani-mals resulted in a dose-dependent elevation in the lipid per-oxidation and depletion of GSH on day 31 postirradiation;both effects were lessened by pretreatment with ZOE. ZOEalso had a dose-dependent antimicrobial activity against Pseu-domonas aeruginosa, Salmonella typhimurium, Escherichia coliand Candida albicans. q 2003 by Radiation Research Society

INTRODUCTION

The first report of the use of chemicals to protect mam-mals against radiation-induced damage appeared in 1949,when Patt et al. (1) reported that cysteine protected miceand rats against radiation-induced sickness and mortality.Since then, several chemical compounds and their ana-logues have been screened for their radioprotective effects.However, the practical applicability of the majority of thesesynthetic compounds has been limited because of toxicityat radioprotective doses (2). Thus it was considered impor-tant to explore alternatives to the synthetic compounds thatwould be radioprotective at nontoxic doses. Plants have

1 Address for correspondence: Department of Radiobiology, KasturbaMedical College, Manipal-576 119, India; e-mail: gc.jagetia@kmc.manipal.edu.

been used to treat various ailments in humans since timeimmemorial, and herbal preparations have usually beenconsidered safer and less toxic than synthetic compounds.Therefore, it is natural that the choices of alternative radio-protectors would include plants and plant products.

The rhizome of Zingiber officinale, commonly known asginger, is consumed worldwide as a spice and flavoringagent. The rhizome of ginger has been reported to possessdiverse medicinal properties in the traditional Indian systemof medicine, the Ayurveda, and it is widely used in severalmedicinal preparations (3–5). Depending on the season, cli-mate, geographic region, and processing, ginger has beenreported to possess terpenes like ar-curcumene, farnesene,b-bisabolene, g-selinene, b-elemene, b-sesquiphellandrene,phellandrene, camphene, a-pinene, b-pinene, cumene, my-recene, limonene, cymene, cineole, citral, zingiberene andbasabolene. Ginger also contains the oxygenated monoter-penes like 2-heptanol, 2-nonanol, n-nonanol, n-decanol,methyl heptenone, 1,8-cineole, borneol, bornyl acetate, lin-alool, geraniol and neral and oxymethyl phenols like gin-gerol, shogaol, zingerone and paradol (4, 6). Most of thesecompounds have been reported to possess antioxidative,free radical scavenging activities (7, 8), to increase anti-oxidant enzymes (9), and to decrease lipid peroxidation(10).

2-Mercaptopropionylglycine (MPG), a synthetic thiolcompound, has been reported to be a radioprotector and hasbeen used in clinics as a detoxifying agent (11). Ocimumsanctum has also been reported to be useful in a numberof therapeutic applications, including antimicrobial and ra-dioprotective activities (12–14). Earlier studies have shownthat an aqueous extract of Ocimum sanctum was most ef-fective at a nontoxic dose of 10 mg/kg once daily for 5consecutive days and protected mice against radiation-in-duced mortality, chromosomal damage, and stem cell dam-age (13, 14).

The common use, wide acceptability by humans, and di-verse medicinal and antioxidative properties attributed toginger rhizome stimulated us to examine the radioprotectiveeffect of the hydroalcoholic extract of ginger (Zingiber of-

585RADIOPROTECTIVE EFFECT OF GINGER

ficinale) in mice after whole-body irradiation with differentdoses of g radiation, using Ocimum sanctum and 2-mer-captopropioonyl glycine (MPG) as positive controls.

MATERIALS AND METHODS

The animal care and handling were carried out according to the guide-lines issued by the World Health Organization, Geneva, and the INSA(Indian National Science Academy, New Delhi). The study was approvedby the Institutional Animal Ethical Committee. Ten- to 12-week-old maleSwiss albino mice weighing 30 to 36 g were selected from an inbredcolony maintained under controlled conditions of temperature (23 628C), humidity (50 6 5%) and light (14 and 10 h of light and dark,respectively). The animals had free access to the sterile water and food.Four animals were housed per polypropylene cage containing sterile pad-dy husk (procured locally) as bedding.

Preparation of the Extract

The ginger rhizomes (small local variety) were collected during Apriland the leaves of Ocimum santum in September 2001. The plants wereidentified by Dr. G. K. Bhat, Department of Botany, Poorna Prajna Col-lege, Udupi, India. The ginger rhizomes were separated, cleaned, freedfrom scales, shade-dried and powdered. The extract of ginger powderwas prepared as described earlier (15). Briefly, 100 g of the ginger powderwas extracted with 50% ethanol at 50 to 608C in a Soxhlet apparatus for72 h. The cooled liquid extract was concentrated by evaporation. Theaqueous extract of Ocimum sanctum was prepared as described earlier(14). The extracts were stored at 2708C until use. The extract of gingerwas characterized by HPLC. The extract of Zingiber officianale and Oci-mum sanctum will be referred to as ZOE and OSE, respectively.

Preparation of the Drug Solution and Mode of Administration

2-Mercaptopropionylglycine (MPG; a kind gift from Santen Pharma-ceuticals Limited, Osaka, Japan) was dissolved in the sterile double-dis-tilled water at 2 mg/ml immediately before use. The pH of the solutionwas adjusted to 6.4 by the addition of 0.1 N sodium hydroxide beforeadministration. OSE and ZOE extracts were dissolved at a concentrationof 1 mg/ml in double-distilled water. The animals were given double-distilled water or drugs intraperitoneally unless stated otherwise and thenwere divided into the following groups:

Double-distilled water 1 radiation. The animals in this group received0.01 ml double-distilled water/g body weight before exposure to g radi-ation.

MPG 1 radiation. The animals in this group were given 20 mg MPG/kg before exposure to 10 Gy g radiation (11).

OSE 1radiation. The animals in this group were injected with 10 mgOSE/kg once daily for 5 consecutive days before irradiation with 10 Gy(14).

ZOE 1 radiation. This group of animals was injected with differentdoses or 10 mg ZOE/kg once daily for 5 consecutive days before expo-sure to radiation (16).

Determination of Acute Toxicity of ZOE

The acute toxicity of ZOE was determined according to Prieur et al.(17) and Ghosh (18). Briefly, the animals were fasted by withdrawingtheir food and water for 18 h and were divided into groups of 10 mice/group. Groups were injected with various doses of freshly prepared ZOE(125, 200, 250, 300, 400, 500, 600, 700, 800 and 1000 mg/kg bodyweight). The animals were provided with food and water immediatelyafter administration of the drug. Mortality was observed for 14 days afterdrug treatment.

Selection of Optimum ZOE Dose

The optimum radioprotective dose of ginger was selected by givingthe animals 5, 10, 20, 40 or 80 mg ZOE/kg once daily for 5 consecutivedays. One hour after the last administration, the animals were exposed to10 Gy of g radiation (16). A minimum of 25 animals were used for eachdrug dose.

Selection of Route of ZOE Administration

The most effective route of administration for radioprotection was se-lected by giving the animals 10 mg ZOE/kg orally or intraperitoneallyonce daily for 5 consecutive days before exposure to 10 Gy of g radiation.A minimum of 25 animals were used for each group.

Selection of Dosing Schedule for ZOE

To select the best schedule for radioprotection, mice were given ZOEas a single dose of 50 mg, 25 mg once daily for 2 days, or 10 mg oncedaily for 5 consecutive days before exposure to 10 Gy of g radiation. Aminimum of 25 animals each were used for each schedule.

Radioprotective Effect of ZOE

To ascertain the radioprotective ability of ZOE, the animals were di-vided into two groups as described above. The animals in the ZOE 1radiation group received 10 mg/kg ZOE once daily for 5 consecutivedays before exposure to 6, 7, 8, 9, 10, 11 or 12 Gy of g radiation. Aminimum of 25 animals were used for each radiation dose in each con-current group.

Irradiation

One hour after the last administration of double-distilled water, ZOEor OSE and 30 min after the administration of MPG, the immobilizedanimals were exposed whole-body to 60Co g radiation (16) in a speciallydesigned well-ventilated acrylic box. Ten animals were irradiated eachtime at a dose rate of 1.66 Gy/min at a source-to-animal distance (mid-point) of 70 cm.

Unless otherwise started, the animals were monitored daily for thedevelopment of symptoms of radiation sickness and for mortality for 30days postirradiation. The dose reduction factor (DRF) was calculated bythe method of Miller and Tainter (19):

LD of ZOE 1 radiation group50DRF 5LD of double-distilled water 1 radiation group50

Estimation of Glutathione and Lipid Peroxidation

The livers of the surviving animals in the ZOE 1 radiation and double-distilled water 1 radiation groups were studied on day 31. Mice werekilled and the livers were perfused with ice-cold saline transcardially.Each whole liver was removed, blotted dry, and weighed, and a 10%homogenate was prepared with an ice-cold 0.2 M sodium phosphate buff-er, pH 8.0, using a homogenizer (Yamato LSG LH-21, Japan). A mini-mum of 4 animals were used for each radiation dose in each group.

Total protein, was estimated by the method of Lowry et al. (20) usingbovine serum albumin as standard, while the GSH contents and lipidperoxidation were estimated as described earlier (21, 22).

Antimicrobial Activity

The antimicrobial activity of ZOE was evaluated on the enteric path-ogens Escherichia coli, Klebsiella, Salmonella typhimurium, Shigella dy-senteriae, Vibrio cholera o1, Pseudomonas aeruginosa and Staphylococ-cus aureus. The antifungal activity of the ginger extract was studied usingCandida albicans, an agent that causes diarrhea in immunocompromisedhosts.

586 JAGETIA ET AL.

FIG. 1. HPLC fingerprints of the 50% hydroalcoholic extract of ginger.

FIG. 2. Kaplan-Meier survival curves for mice treated with variousdoses of ginger, 10 mg/kg Ocimum extract, or 20 mg/kg MPG beforeexposure to 10 Gy g radiation.

The antimicrobial activity was estimated by the agar diffusion method(23). Briefly, the bacterial and Candida colonies were inoculated intopeptone water and nutrient broth and incubated at 378C for 4–6 h. Theturbidity was compared using McFarland’s turbidity standard tube no. 5.Bacteria or fungus were inoculated in sterile petri dishes containing Mull-er-Hinton agar (MHA) and Sabourauds dextrose agar (SDA) by the lawnculture method. Wells of 6 mm in diameter were punched into the agarmedium using a sterile borer. These wells were inoculated with 15, 22.5,30, 37.5 or 45 mg of ZOE. The double-distilled water and solvent controlswere run simultaneously. The plates were incubated at 378C overnight,and the diameters of the zones of inhibition were measured in millimeters.

The statistical significance of differences between the treatments wasdetermined using the Z test for the survival studies, while Student’s t testwas used for biochemical and antimicrobial activities. All the results areexpressed as means 6 SEM (standard errors of the mean).

RESULTS

HPLC Fingerprinting

ZOE was analyzed using HPLC. The chromatogramshows a total of nine peaks; peak 4 corresponds to gingerol,while the other eight peaks represent unknown compounds(Fig. 1).

Determination of Acute Toxicity of ZOE

Administration of ZOE at doses of 125, 200, 250 and300 mg/kg did not induce mortality during the whole ob-servation period. A dose of 400 mg/kg body weight re-sulted in a 20% reduction in the survival of mice, while a50% reduction in survival was observed for 500 mg/kgbody weight, 600 mg/kg resulted in 80% mortality, and100% mortality was observed at doses of 700 mg/kg andhigher.

Selection of Optimum Dose of ZOE

The optimum dose of ZOE for radiation protection wasselected by giving mice 0, 2.5, 5, 10, 20 or 40 mg /kg ZOEbefore whole-body exposure to 10 Gy g radiation. The ad-ministration of these doses of ZOE for 5 consecutive daysdid not induce mortality. The first mortality in the double-distilled water 1 10 Gy radiation group was observed onday 3, and all the animals died by 16 days postirradiation(Fig. 2). Irradiation of mice with 10 Gy induced symptomsof radiation sickness including reduction in food and waterintake, irritability, watering of eyes, epilation, weight loss,

587RADIOPROTECTIVE EFFECT OF GINGER

FIG. 3. Effect of ZOE, OSE (10 mg/kg) or MPG (20 mg/kg) on thesurvival of mice exposed to 10 Gy of g radiation. Solid bars, 10-daysurvival; mottled bars, 30-day survival. IR 5 irradiation, SIR 5 shamirradiation. Error bars are 95% confidence limits.

FIG. 4. Effect of route of administration on the Kaplan-Meier survivalcurves of mice treated with 10-mg/kg ZOE before exposure to 10 Gy gradiation.

FIG. 5. Effect of route of administration on the survival of mice ex-posed to 10 Gy of g radiation. Solid bars: 10-day survival: mottled bars:30-day survival. IR 5 irradiation, SIR 5 sham irradiation. Error bars are95% confidence limits.

emaciation, lethargy, diarrhea and facial edema. The pre-treatment of mice with ZOE delayed or reduced the severityof those symptoms. The onset of radiation-induced mortal-ity was also delayed in the ZOE 1 radiation group com-pared to the double-distilled water 1 10 Gy radiationgroup. The longest delay was observed for 10 and 20 mg/kg ZOE, where the first mortality occurred by day 8 post-irradiation, while the shortest delay was observed for 2.5mg/kg, where the first mortality occurred on day 5 postir-radiation (Fig. 3).

The majority of the animals (67%) in the double-distilledwater 1 10 Gy radiation group died within 10 days afterirradiation. The treatment of mice with ZOE amelioratedgastrointestinal tract (GI) injury as shown by an increasein their 10-day survival. The lowest mortality was observedin the animals treated with 20 mg/kg (12%) followed by10 mg/kg ZOE (25%). A significant elevation in the 10-day survival was observed for treatment with 10 and 20mg/kg ZOE (P , 0.001) treatment (Fig. 3).

Treatment of mice with ZOE resulted in a drug dose-dependent increase in the survival of irradiated animals at30 days up to a dose of 10 mg/kg (P , 0.01), where thehighest survival (33%) was observed (Fig. 3). An increasein the drug dose to 20 and 40 mg resulted in a 13% (P ,0.002) and a 21% reduction in the survival compared to 10mg/kg ZOE (Fig. 2).

Selection of Route of Administration

Greater survival was seen for intraperitoneal than oraladministration (Figs. 4, 5). Therefore, intraperitoneal ad-ministration was used for all subsequent studies.

Selection of Dosing Schedule of ZOE

The daily administration of 10 mg/kg ZOE for 5 con-secutive days (cumulative dose of 50 mg/kg) was found tobe better than a single administration of 50 mg or 25 mg

daily for 2 consecutive days since it afforded greater pro-tection against both the GI and hemopoietic syndromes(Figs. 6, 7). Further studies were carried out using this reg-imen.

Radioprotective Effect of ZOE

The radioprotective action of ZOE was evaluated by us-ing the optimum dose of 10 mg/kg before exposure of themice to 6, 7, 8, 9, 10, 11 or 12 Gy of g radiation. Irradiationof the mice with different doses of g radiation resulted inthe development of symptoms of radiation sickness within2–4 days after exposure. Higher radiation doses resulted inthe early onset of the symptoms. Facial edema was ob-served after 9 Gy or more. Some animals exposed to higherradiation doses exhibited difficulty in locomotion duringthe second week after exposure. The severity of the symp-toms increased with increasing radiation dose.

Whole-body irradiation of mice with 6 Gy did not inducemortality. However, with increasing dose of radiation, sur-

588 JAGETIA ET AL.

FIG. 6. Effect of different dosing schedules on the Kaplan-Meier sur-vival curves for mice treated with 10 mg/kg ZOE before exposure to 10Gy g radiation. IR 5 irradiation, SIR 5 sham irradiation.

FIG. 7. Effect of drug dose schedule of 50 mg/kg ZOE on the survivalof mice exposed to 10 Gy of g radiation. Solid bars: 10-day survival;mottled bars: 30-day survival. IR 5 irradiation, SIR 5 sham irradiation.Error bars are 95% confidence limits.

vival decreased in a dose-dependent manner until a nadirwas reached at 10 Gy, where no survivors were observedbeyond 16 days postirradiation (Fig. 8). As the dose in-creased, the time of mortality advanced (Fig. 8). The LD50/30

was 8.2 Gy for this group. Pretreatment with 10 mg/kgZOE delayed or reduced the severity of radiation sicknessand delayed the onset of mortality by 4–5 days comparedto the concurrent double-distilled water 1 radiation group.Deaths from the GI syndrome were fewer compared to thedouble-distilled water 1 radiation group for all doses (Fig.9a). The number of survivors increased significantly in theZOE 1 radiation group at 30 days postirradiation (P , 0.05for 9 Gy and P , 0.01 for 10 Gy) compared to the con-current double-distilled water 1 radiation group (Fig. 9b).The LD50/30 was 9.4 Gy, an increase of 1.4 Gy, giving adose reduction factor of 1.1 (Fig. 9b).

Glutathione

The administration of ZOE alone before sham irradiationdid not alter the glutathione content. Irradiation resulted ina significant and dose-dependent decline in the GSH con-tents in the double-distilled water 1 radiation group. ZOEtreatment before irradiation elevated the GSH content sig-nificantly (P , 0.01, 0.005) compared to the concurrentdouble-distilled water 1 radiation group.

Lipid Peroxidation

Administration of ZOE before sham irradiation did notincrease lipid peroxidation. Irradiation increased the lipidperoxidation in a dose-dependent manner in both the dou-ble-distilled water 1 radiation and ZOE 1 radiationgroups. Pretreatment with ZOE significantly (P , 0.02,0.001) reduced the induction of lipid peroxidation, therebyprotecting against radiation-induced lipid peroxidation at allthe doses studied. In spite of the reduction in the lipid per-oxidation by ZOE, the lipid peroxidation levels were abovecontrol levels at day 31 postirradiation (Fig. 10b).

Antimicrobial Activity

The zones of inhibition for the alcoholic extract of gingerextract are shown in Table 1. The ginger extract showedlow to moderate inhibitory activity on enteric bacteria andCandida albicans.

DISCUSSION

Dietary ingredients may be very useful if they protectagainst the deleterious effects of ionizing radiation. Gingeris a dietary ingredient that possesses several medicinalproperties (3–5). The aim of the present study was to eval-uate the radioprotective effect of ginger in mice exposedwhole-body to different doses of g radiation.

The exposure of animals to g radiation resulted in radi-ation-induced sickness and mortality; the higher doseskilled all the animals within 10 days, in agreement withearlier reports (14, 16, 24). In mice, death within 10 dayspostirradiation is due to gastrointestinal damage (14, 16,25). The bone marrow stem cells are more sensitive to ra-diation damage than the intestinal crypt cells, but the pe-ripheral blood cells have a longer transit time than the in-testinal cells. Hence the gastrointestinal syndrome appearsearlier than the bone marrow syndrome. In mice, deathfrom 11 to 30 days postirradiation is due to the hemopoieticdamage (14, 25).

The radioprotective effect of ZOE increased in a dose-dependent manner up to 10 mg/kg (once daily for 5 con-secutive days); above that dose, the radioprotective effectdeclined. Earlier studies on radioprotection showed similareffects for other agents (11, 14, 16, 26). The reason maybe that above a particular concentration, a compound maystart manifesting its toxic effects (11, 14, 16, 26). Oral ad-ministration at an equimolar dose did not afford significantprotection. This may be due to the faster absorption of ZOEthrough the i.p. route. A higher oral dose of ZOE may berequired to produce blood levels required for radioprotec-

589RADIOPROTECTIVE EFFECT OF GINGER

FIG. 8. Kaplan-Meier estimate of survival of mice treated with 10 mg/kg ZOE before exposure to various doses of radiation. Panel a: 7 Gy; panelb: 8 Gy; panel c: 9 Gy, panel d: 10 Gy, panel e: 11 Gy, panel f: 12 Gy. Double-distilled water 1 radiation (squares); ZOE 1 radiation (circles).

FIG. 9. Effect of 10 mg/kg ZOE on the survival of mice exposed todifferent doses of g radiation. Double-distilled water 1 radiation (circles);ZOE 1 radiation (squares). Panel a: 10-day survival; panel b: 30-daysurvival. Error bars are 95% confidence limits.

FIG. 10. Alteration in the radiation-induced free GSH contents (panela) and lipid peroxidation (panel b) in the livers of mice exposed to dif-ferent doses of g radiation. Double-distilled water 1 radiation (squares);ZOE 1 radiation (circles). Error bars are 95% confidence limits.

tion, since a significant amount may be lost during the ac-tions of various enzymes in the digestive tract. A similarobservation has also been reported for Ocimum sanctum(14).

Treatment with ZOE once daily for consecutive 5 dayswas superior to the single- or two-dose administration of25 mg for radioprotection, which is in good agreement withthe earlier findings for Moringa oleifera (27). The increasedsurvival of mice after 10 mg/kg ZOE after various radiationdoses indicates the effectiveness of ginger in arrestingdeaths from both the GI and bone marrow syndromes. Thisreduction in death from the GI syndrome may be due to

the protection of the intestinal epithelium. Ginger has beenreported to ameliorate chemotherapy-induced nausea andvomiting and to protect against gastrointestinal hemorrhagein humans (28, 29). Ginger and its active constituents b-sesquiphellendrene, b-bisabolene, ar-curcumene, 6-shogaol,zingiberene, terpenoid, 6-gingerol and 6-gingesulfonic acidhave been reported to inhibit experimentally induced gastriculcerations in rats (30–34). The activities of ginger mayhave helped to protect the intestinal epithelium, allowingbetter absorption of nutrients, and resulting in the prolon-gation of life.

590 JAGETIA ET AL.

TABLE 1Effect of ZOE on the Enteric Pathogens

Microorganism(enteric pathogens)

Zingiber officianale extract (mg/75 ml)

15 22.5 30 37.5 45

Staphylococcus aureusEscherichia coliKlebsiella aerogensSalmonella typhimuriumShigella dysenteriaeVibrio cholera o1Pseudomonas aeruginosaCandida albicans

22212211

12211211

112111

1111

1112

1111

11111

11111

111111

111111

Notes. D 5 diameter of zone of inhibition in millimeters; 111 5 16mm; 11 5 12–15 mm; 1 5 8–11 mm; 2 5 no zone of inhibition.

The hemopoietic syndrome is induced at low doses ofradiation and is manifested by hemopoietic stem cell de-pletion and ultimately by the depletion of mature hemo-poietic and immune cells. The reduction in deaths frombone marrow syndrome by ZOE may be the result of pro-tection of the stem cell compartment of the bone marrow.The LD50 was increased to 9.4 Gy, with a DRF of 1.15.

As far as we are aware, this is the first report of a radio-protective effect of ginger. However, other plants includingPanax ginseng (35), Ocimum sanctum (14) and Mentha ar-vensis (16) have been reported to protect mice against ra-diation-induced mortality. A direct comparison of the pres-ent findings with polyherbal preparations like triphala, cy-stone and abana (36–38) is not possible because of the pres-ence of several plant ingredients, including ginger, in thesedrugs. The present findings are in good agreement with ourearlier findings on mint and Ocimum sanctum, where op-timum radioprotection was observed with a similar doseschedule and route of administration (14, 16).

The observed antimicrobial activity of ZOE is in goodagreement with the earlier reports on ginger (5, 39). Theantimicrobial activity of ZOE may be due to the presenceof terpenes in it, which have also been reported to be an-tibacterial (40, 41). The inhibition of protein, DNA andRNA synthesis and cell wall synthesis and disruption ofplasma membrane are the principal mechanisms of micro-bial killing (42). The antimicrobial activity of ZOE may bedue to one or all of these pathways. Other plants such asOcimum sanctum and mint that show antimicrobial activity(5, 12, 16) have also been reported to be radioprotective.

The mechanism of action of ZOE is not known. Freeradical scavenging is a common mechanism of radiopro-tection. Ginger and its volatile oil have been reported toscavenge free radicals like superoxide anion and H2O2 invitro (43, 44). The aqueous and non-aqueous extracts ofginger along with curcuminoids and cassumunin A and Bthat are present in it have been reported to be antioxidants(45, 46). Unlike most antioxidants and free radical scav-engers, ginger has been found to be anti-inflammatory (47).

One of the peaks in the chromatogram of ZOE corre-sponds to gingerol, which may have contributed to the ob-

served radioprotective action. This possibility is supportedby the observation that gingerol is an antioxidant, an anti-inflammatory agent, and a scavenger of superoxide anion(7, 48, 49). Ginger has also been reported to contain sul-fonated compounds like 4-gingesulfinic and shagosulfonicacids (50), which may also be responsible for observed ra-dioprotection.

Ionizing radiation induces lipid peroxidation, which canlead to DNA damage and cell death (51–53). Therefore, anagent that protects against such damage can provide pro-tection against radiation damage. The administration ofZOE significantly reduced the amount of lipid peroxidationcompared to the concurrent control groups. This inhibitionof lipid peroxidation by ginger extract may also have beenresponsible for the observed radioprotection. Ginger and itsconstituent compound 6-dehydrogingerdione have beenfound to inhibit the induction of lipid peroxidation in vitro(54–58).

The significant elevation in the GSH level by ZOE at allradiation doses may be responsible for the scavenging ofradiation-induced free radicals, including lipid peroxida-tion, and thereby protecting against radiation-induced mor-tality. Ginger has been reported to elevate GSH levels inmice and rats and to reduce lipid peroxidation (55). It hasalso been found to increase the acid-soluble sulfhydryl, glu-tathione-S-transferase and aryl hydrocarbon hydroxylase,superoxide dismutase, catalase and glutathione peroxidasein experimental animals (55, 57). Several investigators havereported that lipid peroxidation starts as soon as the endog-enous GSH is exhausted, and that the addition of GSHpromptly stops further peroxidation (58).

In conclusion, ginger provided protection against radia-tion-induced mortality in mice by protecting against the GIand bone marrow syndromes. The free radical scavenging,elevation in antioxidant status, and reduction in lipid per-oxidation appear to be the important mechanisms of radio-protective action by ginger.

ACKNOWLEDGMENTS

We thank Prof. M. S. Vidyasagar, and Dr. J. Velumurugan, Departmentof Radiotherapy and Oncology, Kasturba Medical College, Manipal, In-dia, for providing the necessary irradiation facilities and radiation dosim-etry. We also thank Prof. P. G. Shivananda and Dr. Mamatha Ballal,Department of Microbiology, for their assistance and help in carrying outantimicrobial studies. We thank Prof. N. Udupa, Dean, and Mr. Subra-manyam, College of Pharmaceutical Sciences, Manipal, for analyzing theginger extract with HPLC. Thanks are also due to Prof. GopalkrishnaBhat Poorna Prajna College Udupi, Karnataka, for identifying the plants.Financial support from the Indian Council for Medical Research, Govt.of India, New Delhi, is gratefully acknowledged.

Received: October 15, 2002; accepted: April 20, 2003

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