Antineoplastic Activity of Crude Saponin Mixture from the Roots ofLuffa tuberosa (Roxb.) in Ehrlich Ascites Carcinoma Bearing Mice

다국어 초록 (Multilingual Abstract)

antitumor activity of crude saponin mixture obtained from Luffa tuberosa (Roxb.) (Fam;Cucurbitaceae) hairy roots (CSLT) in mice transplanted with Ehrlich ascites carcinoma (EAC) was investigated.elevation in tumor-free survival and a highest number of survivors were observed after administration of CSLT(300 .g/kg), which was considered as an optimum dose for its antineoplastic action. The mean survival time(MST) for this dose was approximately 47.1± 0.74 d, when compared with 19.0± 0.36 d of untreated control.Administration of 30 .g/kg CSLT resulted in 130% long-term increased survival time. The measurement ofbody weight, tumor volume, packed cell volume, viable and non-viable count indicated the efficacy of CSLT intumor-bearing mice, there was a significant recovery in hematological profiles, and there was depletion in lipidnormal levels. The CSLT was found to be devoid of conspicuous short-term toxicity in the mice when animalswere intraperitoneally injected with 250, 500, 750 and 1000 .g/kg bodyweight. The treated mice showedconspicuous toxic symptoms only at a dose of 1500 .g/kg. Mortality of the animals was monitored up to 14 dpost drug treatment, 1/7th of the LD50 dose has been considered for the optimal antineoplastic activity.KeywordsLuffa tuberosa (Roxb.) Ehrlich ascites carcinoma, hematological profiles, antioxidant, antineoplasticactivity

참고문헌 (Reference)

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2 "the other group of mice received the normal saline for6 day period and the mice were observed for the toxicbehaviors and life span monitoring Statistical analysisData were expressed as the Mean± S The minimumlevel of significance was fixed at P < 0" -10,

3 "the mice were sacrificed by cervical dislocation rinsed in ice-cold normalsaline solution followed by cold 0 A 10% w/v homogenatewas prepared in 0 15 M Tris-HCl buffer and was used forthe estimation of lipid peroxidation after precipitating proteins with TCA The rest of thehomogenate was centrifuged at 1500 rpm for 15 min at 4oC The supernatant thus obtained was used for theestimation of superoxide dismutase" 19791979

4 "encounteredin the tumor-bearing mice is mainly due to the reductionin the RBC and heamoglobin percentage and this mayoccur either due to iron deficiency or due to hemolytic ormyelophathic condition TheRBC counts and hemoglobin content in the EAC controlgroup was decreased when compared to normal group Treatment with CSLT has increased the RBC count andhemoglobin content close to the normal levels" 1954

5 "countswere measured from freely flowing tail vein blood Differentialleukocyte count of WBC was carried out using Lieshmanstained blood smears for all thegroups of mice" 1965;wintrobeetal19611958

6 "Tumor growth responseThe antineoplastic effect ofCSLT was assessed by change in the body weight and percentageincrease in life span The mean survival time ofeach group of 6 mice was monitored by recording themortality daily for 6 weeks and % ILS was calculatedusing equations Day of first death + Day of last death Mean survival time of treated group/meanSurvival time of control group" Hematological parametersHemoglobin content me : -100, 2006

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8 "The total WBC counts were found to be increased inEAC control group when compared with normal group Administration of CSLT in EAC- bearing mice reducedWBC count significantly as compared with EAC control In the differential count of WBC the neutrophilcount increased where as the lymphocyte count decreasedto a greater extent in EAC control group Treatment withCSLT at diferent doses recovered these altered parametersalmost to the normal values is formed during oxidativedegeneration as a product of free oxygen radicals which is accepted as an indicator oflipid peroxidation the endproduct of lipid peroxidation was reported to be higher incancer tissues than in non-diseased organ" -2, 199019971987

9 "The present study enlights that the crude saponinfraction of L has a remarkable antineo-plastic and antioxidant activity in EAC-bearing mice at aconcentration as low as 150 and 300 These doses selected for the antitumor activity are foundto be optimum by treatment of 24 h old tumors withvarious doses of CSLT which inhibited the body weightgain indicating the arrest of cell proliferation and growth however administration of 400 and 500 g/kg of CSLTwas accompanied by some toxic side effects and thereduction in life span was observed Based on thisobservation the doses of CSLT were selected to 150 and300 g/kg body weights of tumor-baring mice"

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11 "The hairy tuberous root of L tuberosa contains thehigher content of steroidal and triterpenoidal saponins thatare notified by the water solubility A survey of experimentalresults indicates that the saponin mixtures present inplants and plant products possess diverse biologicaleffects when present in the animal body Such as cellmembrane permeabilization as wel the saponinsare widely examined as possible tumor therapeuticsbecause they results in anticarcinogenic effects Most of the experimentsto study the biological effects of saponins have been doneusing crude saponin extracts from plant sources ratherthan purified compounds Because of the presence of alarge number of structurally diferent saponins in plantextracts it is difficult to pinpoint the individual saponinsresponsible for the observed effects" 2005

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13 "The administration of crude saponin mixture of L tuberosa at the dose of 150 and 300 g/kg body weightcaused significant dose-dependent retardation of tumordevelopment and inhibition in the proliferation of carci-noma in Swis albino mice which was evident by theinhibition in the body weight gain increase in life span of the experimental animals to 39 Thegroup treated with the intraperitonial injection of standard5-fluoruracil 47 days of survivaltime Reduction in tumor volume packed cell volume andviable cell count was observed in a dose-dependentmanner in comparison to those of EAC control group nonviable tumor cell counts atdifferent doses of CSLT were increased when comparedwith the EAC control" 73 74 20-0 47 0 0 1,

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15 "Seperation of crude saponin mixtureThe hairyroots of L tuberosa were separated from the plant sliced and dried under shade followed by an incubatorfor 2 d at 40oC crushed in an electrical grinder and thenpowdered wassubjected for extraction with various solvent in soxhletapparatus based on increasing polarity in order ofpetroleum ether methanol and hydroalcoholic mixture of methanol and water the methanol and hydro alcoholic extracts were combinedtogether and diluted with water and fractionated succe-sively with ethyl acetate and n-butanol The resulting n-butanol layer after concentrating was subjected for D 101polymer resin and eluted with water in order to eliminatepolysaccharides A portion of crude saponinafter drying was used to ascertain the antineoplasticactivity" 5 3-,

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19 "Our findings indicated that the thiobarbuturic acid reactivesubstances levels in the tumorous tissues arehigher than those in normal tissues Treatment with CSLTreduced the level of MDA in tumor-bearing mice 1depict the levels of TBARS in liver tissue of experimentalanimals the levels of MDA weresignificantly increased in EAC control animalsAebi Methods of enzymaticanalysis Antimicrobialactivity of certain Sudanese plants used in folkloric Medicine Screening for Antibacterial Activity" 331-337, 19831985

20 "Other cucurbitaceous plants like Momordica charantia have been reported to possessantineoplastic activity in different tumor models in vivoand in vitro Ascites fluid isconsidered to be direct nutritional source for tumor cellsand a rapid increase in ascites fluid with tumor growthwould be a means to meet the nutritional requirement oftumor cells Myelosuppression and anemia are the two majorproblems encountered in cancer chemotherapy" In EAC-bearing mice a regular rapid increasein ascites tumor volume was observed 1995

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22 "Mortality of the animals was monitored up to 14 d postdrug treatment 1/7th of the LD50 dose has considered forthe antineoplastic activity Selection of optimum doseA preliminary experi-ment was carried out to in order to ascertain the dose atwhich maximum anti-tumor activity of CSLT observed For this the EAC-bearing mice were administered CSLTat the doses of 100 g/kg bodyweight once daily for a period of 6" 400-,

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27 "Experimental protocolMale Swiss albino micewere divided in to five groups of twelve animals The CSLT was dissolved in isotonic saline solution and used directly in the assay EACcells were collected from the donor mouse and weresuspended in sterile isotonic saline under the micro-scope and were adjusted at 2× 106 cels/ml 1 ml of EACcells per 10 g body weight of the animals was injected A day of incubation wasallowed for multiplication of the cells" 24-0,

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42 "AnimalsMale Swis albino mice of about 6 to 8weeks of age with an average body weight of 20± 2gwere used for the experiment Indian Institute of Chemical Biology The animals were grouped andhoused in polyacrylic cages and maintained understandard laboratory conditions at a temperature 24± 2oC They were allowed freaccess to standard pellet diet The rats were acclimatized tolaboratory condition for 7 d before comencement ofexperiment Al procedures described were reviewed andapproved by the University Animals Ethical Comittee cellswere obtained from Chittaranjan National Cancer Institute The EAC cells were maintainedin vivo in Swiss albino mice transplantation of 2× 106 cells/mouse after every 10 days and 9 d old EAC cells were used for the screening of theCSLT"

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