“Comparison of anticancer activity of turmeric and mint using Ehrlich Ascites Carcinoma cells”
BRIEF RESUME OF INTENDED WORK
NEED FOR STUDY Cancer is the second largest single cause of death, claiming over six million lives each year worldwide. Depending on the stage of cancer, surgery, radiotherapy and chemotherapy are the most commonly used treatment modalities1.
In the present scenario the search for newer molecules is changing towards the natural bioactive substances owing to their fewer side effects and low cost. About 60% of currently existing anticancer drugs are derived from natural sources2. Turmeric(Curcuma longa) and Mint(Mentha Piperita) are frequently used in our diet. Anticancer activity of these two functional foods has been proved 3,4. The present study is taken up to compare the anticancer activity of turmeric and mint in the EAC tumor bearing mice as per the standard protocol recommended by the National Cancer Institute, USA. This will enable us to suggest to increase the frequency of the more potent of the two in diet for prophylactic/curative purpose. Review of Literature:
Herbal drugs constitute a major share of all the officially recognised systems of health in India viz. Ayurveda, Yoga, Unani, Siddha, Homeopathy and Naturopathy, except Allopathy. More than 70% of India’s 1.1 billion populations still use these non-allopathic systems of medicine. Currently, there is no separate category of herbal drugs or dietary supplements, as per the Indian Drugs Act. Significant basic and clinical research has been carried out on the medicinal plants and their formulations, with the state-of-the-art methods in a number of Institutes/Universities5. Turmeric is dried as well as fresh rhizomes of the plant known as Curcuma longa Linn. Belonging to the family Zingiberaceae. It contains about 5% of volatile oil, resins8. In traditional Indian Ayurvedic medicine, turmeric has been used as a tonic for the digestive system and the liver; to dispel worms, strengthen the body, and dissolve gallstones; and for menstrual irregularity and arthritis. In old Hindu texts it is described as an aromatic, stimulant, and carminative (an agent that helps expel gas from the intestines and treats colic). Mixed with slaked (hydrated) lime, turmeric was a well known household remedy for sprains and swellings caused by injury7. It has been proved to posses anti-inflammatory, antiarthritic6. antioxidant, antifungal8 and anticancer activities3,6. Mint a widely available herb with a number of medicinal properties and helps to promot good health. The oil is obtained by steam distillation of the fresh flowering tops of plants known as Mentha piperita Linn. Belonging to the family Labiatae. It contains volatile oils majorly l-menthol. Mentha oil is traditionally used as carminative, stimulant and flavouring agent6. It proved to posses antiseptic, antimicrobial9 and anticancer activities10. Objective of the study:
The objectives of the present study are as follows :
Comparison of anticancer activity of turmeric and mint, using EAC.
Evaluation of antioxidant enzyme in turmeric and mint pretreated mice.
MATERIALS & METHODS Source of data
Whole work is planned to generate data from laboratory studies i.e.; experiments are performed as described in reference, experimental studies in journals and in textbooks available with college, IISc library, RGUHS digital library (Helinet), websites www.sciencedirect.com,
www.ncbi.nlm.nih.gov/pubmed,www.google.com,www.ijp-online.com,will be used to obtain related information regarding this research protocol.
METHODS OF COLLECTION OF DATA
The data collected will be based on laboratory animal experimentation 1. Preparation of drug and mode of administration:
The drugs Mentha piperita and Curcuma longa will be obtained locally and authenticated. The crude drug will be shade dried, powdered and extracted using distilled water.
Swiss albino mice either sex of 6-8 weeks old (25±5 g body weight) will be used in these study. The animal experiments will be performed according to the rules and regulations of the Institutional Animal Ethics Committee1,11. 3.Tumor cells
The Ehrlich’s Ascites Carcinoma (EAC), will be obtained from, Amala Cancer Research Center, Thrissur, Kerala. They will be maintained by weekly intraperitoneal inoculation of 106 cells/mouse.
4.Anticancer effects in the Ehrlich Ascites Carcinoma bearing mice
The preinoculated (1x106 cells/mice, i.p.) will be maintained for 15 days12,13. After two week of inoculation, the peritoneal fluid will be aspirated and diluted to get 1x106 EAC cell / ml in a phosphate buffer. Mice will be reinoculated with 1x106 EAC cell / mice 12 and divided into 5 groups consisting of 8 animals in each group.
No. of Animals
EAC mice+ distilled water
EAC mice + cisplatin
EAC mice + turmeric
500mg/kg b.wt 15.
EAC mice + mint
b. wt = body weight
Group 1-5 will be treated with vehicle & drug samples orally 24 hrs after the inoculation of EAC and continued for 9 days and the observation will continue until the termination of the study.The dose of Cisplatin, turmeric and mint are selected based on literature.
TUMOR GROWTH RESPONSE1
At termination, surviving animals of EAC tumor bearing mice will be counted and the average survival time (MST)17 and the % increase in life span (% ILS) will be Calculated by following formula.
Median survival time (MST) = (Day of 1st death + Day of last death) / 2
Percentage increase in life span = [ (MST of treated group/ MST of control group) – 1] x 100 Body weight analysis: During the same time of study, weekly body weight analysis will be recorded.
% Increase in weight = [(animal weight on respective day/animal weigh on day 0) – 1] x 100 ESTIMATION OF HEMATOLOGICAL PARAMETERS1
On the last day of treatment, blood will be drawn from each mouse in the conventional way and the white blood cell count (WBC), red blood cell count (RBC), hemoglobin, differential count will be determined using Haemocytometer and Sahli’s haemometer. ESTIMATION OF ANTIOXIDANT ENZYME18.
Lipid peroxidase , glutathione, super oxide dismutase and catalase levels will be measured in tumor cells collected from untreated mice and 1 h after treatment of tumor-transplanted mice with last dose in each group by using standard procedures. STATISTICAL ANALYSIS:-
The data obtained from experimentation will be subjected to one way Analysis Of Variance (ANOVA) with suitable post-hoc test.
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