Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524111EnglishN-0001November30HealthcareALTERNATIVE TO USE OF LIVE ANIMAL IN TEACHING PHARMACOLOGY AND PHYSIOLOGY IN PHARMACY UNDERGRADUATE CURRICULUM: AN ASSESSMENT OF 120 STUDENTS VIEWS
English0309S.V. TembhurneEnglish D.M.SakarkarEnglishThis study was conducted to ascertain the attitudes of undergraduate pharmacy students to animal experimentation after they had completed experiments on frog abdominal rectus muscle, goat intestine, studied the effect of various drug like analgesic, anti-inflammatory, diuretics etc on mice and rats and on computer programme in academic year 2008-09. Barely sixty five percent of the students agreed with to use alternative of life animals like mice and rats for purpose in education, an overwhelming majority (seventy four percent) felt that the animal experiments involved needless pain and suffering to the animals. A large majority of students felt the need to reduce the number of animals by demonstrating the practical, to explore alternatives to the animal experiments, and to restrict animal experiments for research on life threatening diseases. More than half of the students were unaware of the government regulations on animal experiments. Students considered, understanding physiological processes as the main objective of animal experiments. The results of this study indicate that there is a need to modify the curriculum taking into account the development and availability of the new technology. The students also need to be made aware of the Government regulations on animal experiments particularly the role of the CPCSEA and the institutional animal ethics committee.
EnglishAnimal experiments, Physiology, medical education, undergraduate, ethics.INTRODUCTION
Pharmaceutical Undergraduate curriculum in India continues to follow an archaic syllabus set up decades ago, which has not kept pace with the technological progress. The Animal Physiology syllabus does not reflect the constraints imposed by the Prevention of Cruelty to Animals act of 1960 in the transport, care and experimentation. The CPCSEA (Committee for the Purpose of Control and Supervision of Experiments on Animals) was established under Prevention of Cruelty to Animals act of 1960. The Experiments on Animals (Control and Supervision) Amendment Rules (1998) and the Breeding of and Experiments on Animals (Control and Supervision) Rules (1998) gave powers to the CPCSEA, to make rules in relation to the conduct of experiments on animals, the power to authorize any of its officers to inspect any place (at any time) and the power to prohibit a person or an institution from carrying out experiments on animals1.
In the pharmaceutical undergraduate curriculum especially in experimental pharmacology a lot of stress is laid on pithing. It is usually performed by inserting a sharp probe into the living animals spinal cord or brain, through the nape of the neck, moving the probe vigorously to destroy the brain. It is a common site to see a hapless frog jumping in the lab with a needle passing through its eye or other parts to the shrieks of second and third year students of pharmacy. It is difficult to see the rationale of such rituals in making a good pharmacist.
In the decade since, several states of USA have passed "choice-in-dissection" laws, which affirm a students right to use alternatives to dissection without penalty. In the past, the use of live animals has been routine practice in the pharmacy undergraduate training curriculum. However, recent trends indicate that animal use is declining. According to the Physicians Committee for Responsible Medicine (PCRM), which for the past decade has been pressuring medical schools to replace animal labs with non-animal alternatives, about half of all 126 U.S. medical schools-including prestigious institutions as Mayo, Harvard, Columbia, and Yale-now have no live animal laboratories. One clear conclusion that can be drawn from this information is that live-animal use is not indispensable for pharmacy training also2 .
Keiser and Hamm argue that when dissection is not part of the school curriculum, students may miss the opportunity to prepare for vocations and become valuable contributors in medicine and other health-related professions. However, many students switch career plans away from the life sciences when they learned that they were required to dissect animals3 . There are small number of studies have been conducted in alternative to use of animal in undergraduate training curriculum for saving the live of animals. The study can be also performing on goat intestine which can be explained by our previous results on goat intestine4-6 . Bennett conducted study on 110 U.S. Medical students. 78 percent of the surveyed medical students supported a students right to choose not to participate in required terminal dog labs, and 32 percent felt that, given a choice, they would not participate in such labs7 . Bowd conducted 191 Canadian Undergraduates In a retrospective survey, 27 percent of the surveyed students reported having exclusively negative reactions to dissection, and 38 percent reported both negative and positive reactions8 .
Many studies in the West have captured the pitiable conditions in which the lab animals are kept. Gibbs et al. conducted an in-depth study to document the conditions of the capture and warehousing of frogs bound primarily for dissection. "As many as 100 frogs were kept in each sack for up to a week or more, the only care being intermittent spraying with water. Eventually, the frogs were put into large tubs of water where they were kept for periods ranging from days to months depending on the season and the demand for shipments. During this period, the frogs were provided no food. Frogs shipped during the summer likely had gone without food for a week or more between capture and arrival at a school; in the early spring, frogs may not have eaten for more than six months. In the summer months, most frogs were "hot," meaning that they were overheated and hyperactive often to the point of convulsion9 ." disciplines, to 39 percent, 10 percent, and 17 percent11 .
MATERIALS AND METHODS
The survey was conducted among the undergraduate second and third year pharmacy students at S.N.Institute of Pharmacy, Pusad during academic year 2008-09. The 120 students had been exposed to various animal experiments e.g. experimentations on frog abdominal rectus muscle, goat intestine for absorption and bioassays studies, effect of various drugs like analgesic, antiinflammatory, diuretics etc and various surgical techniques like adrenoctomy and ovarectomy in mice, rats, while undergoing a practical syllabus of pharmacology subject of Pharmacy second and third year. All the experiments were performed by taking a permission of Institutional Animal Ethical Committee of S.N.Institute of Pharmacy, Pusad. The study was conducted using a questionnaire. The questionnaire was filled anonymously, and voluntarily. Opinion was sought on the following aspects, the need to use animals, alternatives to animal experiments, awareness of government regulations and the objectives of animal experiments.
RESULTS AND DISCUSSION
The CPCSEA (Committee for the Purpose of Control and Supervision of Experiments on Animals) was established under Prevention of Cruelty to Animals act of 1960. The Experiments on Animals (Control and Supervision) Amendment Rules (1998) and established 4 Rs i.e. reduction, refinement, replacement and rehabilitation in relation to the conduct of experiments on animals1, 12 .
Reduction: to constantly work to reduce the number of animals used in research.
Refinement: to improve the lives and living conditions of animals used in research, to make their lives comfortable.
Replacement: to constantly try to find new and better ways to do research that do not require animals.
Table1: Students views toward the use of animals in their practical curriculum
Values given in parenthesis are present in the form of percentage.
There is a global trend towards reduction in animal experiments medical undergraduate training similarly there is needs to follow such trend in pharmacy undergraduate training. The results of this study indicate that there is a need to modify the curriculum taking into account the development and availability of the new technology to use of alternative to live animals13 . In present study 120 undergraduate pharmacy students responded to the questionnaire. The results are summarized in Table 1. Barely sixty four percent of the students disagreed with to use of frog for studying bioassays and other pharmacology experiments in pharmacy curriculum while seventy eight percent of students felt alternative use of goat intestine for bioassay and for other experiments in pharmacology practicals.
An overwhelming majority sixty five percent students felt alternative to use of live animals like computer programs, video films, demonstration by teacher for teaching pharmacology and physiological experiments. A majority of students (forty nine) felt to reduce the number of animals by demonstrating the practical, to explore alternatives to the animal experiments, and to restrict animal experiments for research on life threatening diseases; seventy four percent of students felt the animal experiments involved needless pain and suffering to the animals and these are might be reasons for the sixty five students views toward alternative to use of live animals in teaching. While twenty nine percent of students felt to use of live animals in teaching and this was might be due to thinking the use of animals for better understanding of physiology, improving dissection skill and for passing the university examination.
More than fifty percent of the students were unaware of the government regulations on animal experiments. Students considered, understanding physiological processes as the main objective of animal experiments. Thus students need to make aware of the Government regulations on animal experiments particularly the role of the CPCSEA and the institutional animal ethics committee. There are many alternatives to teach the pharmacological experiments. Computer simulations allow students to view many levels of complexity unavailable to the dissector.
Programs currently being used The CD-ROMs on the human body produced by ADAM (Animated Dissection of Anatomy for Medicine) software, for example, show not only gross structural anatomy in high detail, but also contain histology images, animations, and video clips of body processes unobservable during gross dissection of a living or dead organism. Digital Frog by Digital Frog International. 3-D Body Adventure, by Knowledge Adventure displays "fly through" of the skeletal and circulatory systems of the human, in which the viewer tours these systems in threedimensional space as if piloting a miniature airplane. Physiology is one of the heaviest users of animals. A survey by the Association of Chairmen of Departments of Physiology reported that most physiology faculty believed that no alternative could fully replace liveanimal use in education14 . The Virtual Physiology Series (five CD-ROMs), produced at the University of Marburg, Germany, covers the entire field of nerve-muscle physiology and simulates all of the classic experiments15 .
The SimBioSys Physiology Labs use animations, simulations, exercises, and quizzes, and cover general, cardiovascular, respiratory, and renal physiology; over 1,000 physiological parameters can be reproduced; by altering parameters, students gain understanding of how the body works16 . Dyna Pulse Systems allows students to monitor their own cardiovascular profiles; also includes a "patient management" system that allows longterm tracking and statistical analyses of students cardiovascular status17 . Intelitools software series allows students to study respiratory physiology (Spriocomp), muscle contraction (Physiogrip, Flexicomp), and cardiac physiology (Cardiocomp); students generate their own original data from their own bodies, making them both the investigators and the experimental subjects18. To sum up there is no dearth of alternatives.
CONCLUSION
The results of this study indicate that there is a need to modify the curriculum taking into account the development and availability of the new technology. The students also need to be made aware of the Government regulations on animal experiments particularly the role of the CPCSEA and the institutional animal ethics committee.
Englishhttp://ijcrr.com/abstract.php?article_id=2274http://ijcrr.com/article_html.php?did=22741. The Breeding of and Experiments on Animals (control and supervision) Rules. The Gazette of India no.809, 1998.
2. Physicians Committee for Responsible Medicine (PCRM). 1998. Medical school Curricula with no live animal laboratories. Washington, D.C.: PCRM, 1998.
3. Keiser TD and Hamm RW. Forum: Dissection-The case for. The Science Teacher 1991; 58(1): 13-15.
4. Tembhurne SV, Sakarkar DM. Transport of Glucose in isolated goat ileum preparation: an alternative research tool for absorption study. Indian J. Physiol Pharmacol 2008; 52(2): 211-13.
5. Dewhurst DG, Hardcastle J, Hardcastle PT, Stuart E. Comparison of a computer simulation program and a traditional laboratory practical class for teaching the principles of intestinal absorption. American Journal of Physiology 1994; 267: S95-S104.
6. Surendra H. Bodakhe, J.S. Dangi, Alpana Ram, K. P. Namdeo and Kiran S. Bodakhe. Isolated Cock Ileum: A Tool for Pharmacology Experiments. Indian J.Pharm. Educ. Res. 43(2), 2009, 199-202.
7. Bennett J. New survey shows Colorado students want a choice. Good Medicine 1994; 3(3): 6.
8. Bowd AD. Dissection as an instructional technique in secondary science: Choice and alternatives. Society and Animals. 1993; 1(1): 83-88.
9. Gibbs EL, Nace GW, Emmons MB. The live frog is almost dead. BioScience 1971; 21: 1027- 34.
10. Schrock JR. Dissection. The Kansas School Naturalist 1990; 36(3): 3-16.
11. Ammons SW. Use of live animals in the curricula of U.S. medical schools in 1994. Academic Medicine 1995; 70: 740 43.
12. Guidelines for care and use of Animals in Scientific Research, Indian National Science Academy, 2000.
13. Barnard ND, Stolz J, Baron L. Use of and alternatives to animals in laboratory courses at U.S. medical schools. Journal of Medical Education 1988; 63: 720-22.
14. Greenwald GS. ACDP survey on use of animals in teaching physiology. Physiologist 1985; 28: 478- 80.
15. Thieme Interactive. Virtual physiology: The unique truly interactive simulation Software Promotional brochure. (http://www.thieme.com)
16. Critical Concepts, Inc. (CCI). Critical Concepts releases physiology simulation on CDROM: Academia can save thousands using computer simulations. Company press release, 15 April.1999. (http:/ /www.laketech.com)
17. Pankiewicz PR. Software review: The DynaPulse 200M. The American Biology Teacher 1995; 57(2): 121-22.
18. Intelitool Physiogrip and Spriocomp: Capabilities and features, Promotional brochures 1998. (http://www.intelitool.com)
Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524111EnglishN-0001November30HealthcareSTUDYING PHYSICIANS PERCEPTION TOWARDS MEDICAL REPRESENTATIVES
English1013Nidhi SinhaEnglish Ajay PiseEnglish Shilpa PiseEnglish N. UdupaEnglishEnglishINTRODUCTION
Concept of perception Perception is the process by which organisms interpret and organize sensation to produce a meaningful experience of the world. It describes the ability to understand the true nature of something. In humans it is a process whereby sensory stimulation is translated into organized experience.
Importance of medical representatives in pharmaceutical marketing Medical representatives are the backbone of any pharmaceutical company. They are the link between the company and the physicians. It is his responsibility to convince the prescribers to prescribe the products of his company. For this he offers literatures, gifts, and free product samples to the prescriber. They provide the physicians with scientific and clinical information. It is his duty to keep the physicians updated about new products, the purpose for which it is intended to be used, the possible side effects of it and all other prescribing information. They also keep them informed about the ongoing research in his company. It is he who enquires the physician about his experience in using the companys product and actions to be taken in case of any complaints. He can also act as an advisor to the physician in case any adverse drug reaction is reported.
Changing trends in sales promotion of pharmaceutical marketing Promotional activities for product are considered as life blood of all marketing strategies. It helps in increasing the sale of drug. In pharmaceutical marketing, product promotional strategies are mainly focus on inducing physicians prescription. In todays scenario Pharmaceutical companies are facing cut-throat competition and they are applying several new methods of product promotion to promote their brand. In this process, they spend heavily on advertising which leads to increased price of the product. Pharmaceutical companies are promoting their product through medical representatives, journals and direct emailing to the physicians.
OBJECTIVES OF STUDY
1. Analyzing physicians perception towards medical representatives
2. To identify the nature and extent of relationship between the physicians and the medical representatives and its effect on the behavior of physicians
3. To identify and analyze the influencing factors in medical representatives
4. To analyze effectiveness of verbal communication of medical representatives against the visual aids
5. To identify whether the drug information from internet sources can replace the medical representatives
6. To analyze which age group of medical representatives are most efficient
7. To analyze whether female medical representatives are more influencing than the male
IMPORTANCE OF STUDY: This study is important in the following areasThe companies train their medical representatives in a way which is beneficial to them. This study will help the company to strive and change the strategy of training of medical representatives to increase the overall productivity of the company. This study will help the physicians to gain maximum benefit from the medical representatives regarding their knowledge of drugs within short span of time. The physicians have a very busy schedule and they can hardly afford to spend more than 5 minutes on each medical representative. With the help of this study the medical representatives will aim to deliver the physicians only the required information which is expected out of him. Thus he will try to make the best possible use of the physicians precious time. This study is also important for academic purpose as it will reveal and encourage further studies to be carried out in this area.
METHODOLOGY:
1. Determining sample size by application of market research software (statistics calculator software). a. Best estimate of the population size in the area (physicians) = 1500 b. Best estimate of the rate in the population (%) = 50 c. Maximum acceptable difference (%) = 7 d. Desired confidence level (%) = 85 e. Required sample size = 98
2. Preparation of questionnaire: Open and close ended questions were prepared to analyze the perception of physicians regarding medical representatives.
3. Sampling unit: The prepared questionnaire was administered from tertiary care hospitals, clinics, nursing homes and govt. hospitals in the area of North Karnataka state of India
4. Data analysis: Percentage analysis method was used to analyze the collected responses.
RESULTS AND DISCUSSION: It was observed that 70.4% physicians spend less than 5 min with each medical representative. 74.5% physicians were of the opinion that they attend 1-5 medical representatives daily. The maximum range which a physician attended was 6- 10. Good communication skill was considered as the influencing factor by 48.9% physicians; whereas knowledge was given high ranking by 24.1% physicians. A few of them believed positive attitude and personality to be significant. 59.2% physicians opined that medical representatives provide ample information about new products. 53.1% physicians denied that medical representatives provide sufficient product samples. 50% physicians believed verbal communication to be more effective than their visual aids. In such a modern technological era 47.95% physicians alleged that drug information from internet sources cannot replace the medical representatives, whereas 33.7% believed that both of them are required for them to be updated regarding drug information. 67.3% physicians recommended that medical representatives should change their detailing style on subsequent visits. 61.2% physicians were of the opinion that medical representatives were not able to clear their doubts about the products. 26.5% believed that they were able to do so only on their next visit. Regarding the age, 59.2% physicians were of the opinion that medical representatives of age group 26-30 were the best in their work. 73.5% physicians believed that female medical representatives are less influencing than their male counterpart.
CONCLUSION: From above study it was observed that majority of physicians (70.4%) spend less than 5 min with each medical representative, it indicates that medical representative has to convince physicians within less than five minutes. 74.5% physicians attend 1-5 medical representatives daily. As good communication skill was considered as the influencing factor by 48.9% physicians, pharmaceutical companies can focus their training mainly on communication skills for their sales force. It is observed that 53.1% physicians denied that medical representatives provide sufficient product samples, by taking this in account we can conclude that pharmaceutical companies can provide more samples to physicians.
Above study shows that 50% physicians believed verbal communication of Medical Representative are more effective than visual aids; it indicates that pharmaceutical companies should concentrate more on developing verbal communication of medical representatives. Majority of physicians (67.3%) recommended that medical representatives should change their detailing style on subsequent visits. 61.2% physicians were of the opinion that medical representatives were not able to clear their doubts about the products at first visit. Majority of physicians were of the opinion that medical representatives of age group 26- 30 are the best in their work. 73.5% physicians believed that male medical representatives are more influencing than their female counterpart.
RECOMMENDATIONS: From above conclusion we would like to recommend-
1. Medical Representative has to convince physicians within less than five minutes, therefore he has to put his efforts to convince physicians within short span of time.
2. Pharmaceutical companies should train their medical representatives in verbal communication.
3. Medical representatives should change their detailing style in every subsequent visit.
4. Medical representatives should possess detail knowledge about the product so that they can clarify physicians doubt on the spot.
LIMITATIONS OF STUDY: The sample size considered in this study is restricted to 98. It is not universal representation; therefore results derived from this study should not be generalized.
Englishhttp://ijcrr.com/abstract.php?article_id=2275http://ijcrr.com/article_html.php?did=2275Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524111EnglishN-0001November30HealthcareANALYZING MEDICAL REPRESENTATIVES ROLE IN PRESCRIPTION WRITING HABIT OF THE PHYSICIANS
English1415Preeti KumariEnglish Ajay PiseEnglish N. UdupaEnglishStudy was designed to identify the role of medical representatives in prescription writing habit of the physicians. Identifying the source of information of ADR and to identify the role of hospital pharmacy in the prescription writing habit were secondary objectives of the study. This study is important for Medical Representatives because it gives them the knowledge bout the factors which influences the prescription writing habit. This study mainly focuses on analysis of physicians’ prescription writing behaviour. This analysis would help companies to understand physicians’ mindset and to target company’s marketing efforts precisely. A questionnaire designed with open and close ended questions to collect information against the framed objectives. A market research software (statistics calculator) was used to determine sample size from the population. Convenient sampling process was followed. Government hospitals, private hospitals, private clinics, medical college from , Udupi, Mangalore and nearby area. Collected data was analyzed by using percentage analysis method to derive results.
EnglishObjectives of study
1. To identify the role of medical representatives in prescription writing habit of the physicians
2. To know the pattern of prescription for different types of patients
3. To identify the role of hospital pharmacy in the prescription writing habit of the physician
4. To identify the priority of dosage form in the prescription
5. To identify the source of information of ADR
Importance of study:
1. Medical Representative: Medical Representative is the direct source of information for physician about the drug. This study is important for Medical Representatives because it gives them the knowledge bout the factors which influences the prescription writing habit of physician.
2. Pharmaceutical Industry: This study mainly focuses on the analysis of physicians’ prescription writing behviour. This analysis would help pharmaceutical companies to understand physicians’ mindset and to target company’s marketing efforts precisely.
3. Hospital pharmacy: As the prescription writing habit of physician are greatly influenced by the medicine present in hospital pharmacy. Hence this study will help the hospital organization to implement some policies for betterment of hospital pharmacy.
Research methodology: Questionnaire design: A questionnaire was designed with open and close ended questions to collect information against the framed objectives. Designing sample size: A market research software (statistics calculator) was used to determine sample size from the population. Total estimate of physicians’ population in selected area: 1000 Best estimate of the rate in the population: 50% Maximum acceptable difference: 9% Desired confidence level: 95% Required sample size: 105
Sampling process: convenient sampling Sampling Unit: Government hospitals, private hospitals, private clinics, medical college from Manipal, Udupi, Mangalore and nearby area Data Analysis: collected data was analyzed by using percentage analysis method.
Results: Among 187 responses from physicians most (24.06%) of the doctors think that internet is the best source for updating their knowledge about the new drug. 21.93% respondents believe that journals are good source of information for updating their knowledge. 20.86% physicians update their knowledge from the information provided by Medical representatives. Respondents also update their knowledge about drug from text books (15.5%), CIMS(10.16%), Conferences(4.23%), MIMS(3.21%).
Englishhttp://ijcrr.com/abstract.php?article_id=2276http://ijcrr.com/article_html.php?did=2276Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-524111EnglishN-0001November30HealthcareINFLUENCE OF BENZENE EXTRACT OF PIPER BETEL ON DELAYED SMALL INTESTINAL TRANSIT BY
CHLORPROMAZINE INVOLVING CALCIUM INNERVATIONS IN MICE
English1622Bairagi G.B.English Hatkar A.D.English Zaferuddin S.D.English Dhaked P.S.English Tembhurne S.V.English Sakarkar D.MEnglishPiper Betel Popularly known as ‘Pan’ in India belongs to family Piperaceae. Medicinally it has stimulant, antiseptic, sialogogue activity. It is reported that calcium is involved in the initiation of contraction of smooth muscle. It increase intestinal motility through L-Type. Chlorpromazine blocks the calcium channel on smooth muscle and relaxes by attenuating intestinal motility. The present study was to evaluate the influence of benzene extract of Piper betel (BEPB) on small intestinal motility (SIT). The study was also undertaken to evaluate the mechanism involve in SIT. BEPB (400 and 800 mg/kg p.o) administered to 15 hrs fasted Swiss albino mice. 4% charcoal meal was administered (10 ml/kg p.o) 1hr after the drug administration and after 20 min all animals were dissected for determination of SIT. For exploration of calcium channel in gastrointestinal motility chlorpromazine (5mg/kg p. o.) was administer 30 min prior administration of drug respectively. The results of study indicate that BEPB accelerate the intestinal transit in normal mice. At 800mg/kg dose it was found to accelerate the transit by 89.5% compared to normal mice which was significant at (P < 0.05) compared to vehicle group. Chlorpromazine inhibits GI transit by 28.85% in normal mice. In presence of BEPB it able to reverse the effect ofchlorpromazine, while the significant effect was observed only at 800mg/kg BEPB. In presence of BEPB chlorpromazine could able to produce upto15.85% inhibitions of SIT which was comparable to standard Metaclopramide (10mg/kg p.o) indicates BEPB could partly produce acceleration effect through calcium involvement as well as by some other pathway as chlorpromazine could not completely inhibit the acceleratory effect of Piper betel on small intestinal transit.
EnglishPiper betel, Intestinal transit, Chlorpromazine, Calcium channel.INTRODUCTION Gastrointestinal dysmotility impacts on the quality of life of patients for example, a significant percentage of patients with diabetes have gastrointestinal dysmotility. Gastrointestinal complications of diabetes can affect one or more parts of the gut and produce nausea, vomiting, abdominal pain, constipation and/or diarrhea. Abnormal gastric emptying, or gastroparesis, may lead to poor glucose control and complications of diabetes 1, 2, 3 . The gastrointestinal tract is in a continuous state of contraction, relaxation and secretion. These functions are controlled by neurohumoral systems, which in turn are regulated by various receptor systems, such as cholinergic, adrenergic, serotonergic, opioidergic and calcium channels. Many drugs affect GI transit by acting as agonists or antagonists at specific cellular receptors, such as cholinergic4 , adrenergic5 , serotonergic6, 7, opioidergic8, 9 , and calcium channels10, 11 .
In present study we used Piper Betel for screening of gastrointestinal motility. Piper Betel popularly known as ‘Pan’ in India belongs to family piperaceae. In India Peoples are widely using this after having meals for digestion and warm taste, medicinally it has stimulant, antiseptic, sialogogue activity12. It is reported that calcium is involved in the contraction of smooth muscle. It increases small intestinal motility through L-Type channel10, 11. Thus the present study was designed to investigate the effect of benzene extract of Piper betel on small intestinal motility. The main objective of the present study was to evaluate the involvement of Ca2+ in acceleration of intestinal transit time by using chlorpromazine which blocks calcium channel on smooth muscle cell and relaxes by attenuating intestinal motility.
Material and Method Preparation of Extract: The fresh leaves of Piper betel were collected from local market at village Pusad in Yavatmal district region, Maharashtra, India. The collected leaves of Piper betel were dried under shade and undergone crushing in electric blender to form powdered and subjected to extraction by using Maceration in a air tight closed container by using benzene as a solvent. The extract was concentrated by evaporation at room temperature and used for testing a gastrointestinal motility.
Material: benzene extract of Piper betel leaves, Activated charcoal (S.D. Fine chemical, Mumbai) and Chlorpromazine (La Pharmaceutical Pvt. Ltd., Ahmedabad), Metaclopramide (Wallace Pharmaceutical Ponda,Goa.)
Experimental animals: All the experiments were carried out in adult Swiss albino male mice. The animals were fasted for 15 hrs prior experimentation while had free access to water, and they were housed in a natural (12 hrs each) light–dark cycle. The animals were acclimatized to the laboratory conditions for at least 5 days before exposed for experimentation. The experimental protocol was approved by the Institutional Animal Ethics Committee (IAEC) and the care of laboratory animals was taken according to the guidelines of CPCSEA, Ministry of Forests and Environment, Government of India (registration number 729/02/a/ CPCSEA).
Administration of Extract: Suspension of Benzene extract was prepared in 0.5% carboxymethyl cellulose using tween 20 (0.2% v/v) as a suspending agent. The extract was administered in a dose of 400 and 800mg/kg p.o. respectively. Control groups were given only 0.5% carboxymethyl cellulose with tween 20 (0.2% v/v).
Administration of charcoal: The mice were administered charcoal meal consisting of 4 % of activated charcoal and 2% carboxy methyl cellulose orally (10ml/kg) after 1 hr. of respective treatments.
Administration of Chlorpromazine: The mice were administered with Chlorpromazine (5mg/kg p.o.)30 min prior treated with drugs.
Administration of Metaclopramide: The mice were administered with Metaclopramide (10mg/kg p.o.) as similar with test extract.
EXPERIMENTAL DESIGN Mice were randomly divided into 7 groups of 5 animals each. All the animals were fasted for 15 hrs. Group 1 served as a control and received vehicle only. Group 2, 3 were selected for evaluating acceleratory effects of Piper betel on intestinal transit while group 4, 5, 6 were selected for assessing the calcium channel in acceleration of intestinal transit by using chlorpromazine while group 7 was selected as standard Metaclopramide.
Acceleratory effect of Piper betel on GI transit in normal mice: For evaluation of acceleratory effect on intestinal transit, group 2, 3 received benzene extract of Piper betel (400 and 800mg/kg p.o) respectively.
Statistical Analysis All value are expressed as the mean ± S.D. Statistical significance was assessed by the unpaired Student’s t test for all results.
RESULTS
Acceleratory Effect of Piper betel on Intestinal Transit Benzene extract of Piper betel administration at higher doses (800 mg/kg) produced a significant (P < 0.05 Table 1) acceleration of intestinal transit while at lower dose (400 mg/kg) unable to produce significant effects (Table 1 and Figure 1).
Calcium channel: Influence of Piper betel on delay transit by Chlorpromazine 30 minute prior treatments, group 4, 5, 6, 7 of animals were treated with Chlorpromazine (5 mg/kg p.o.) for induction of delayed intestinal transit. In which group 4 served as a pure Chlorpromazine treated group for evaluating calcium channel in induction of delaying transit time, while remaining groups received benzene extract of piper betel (400 and 800mg/kg p.o) and standard Metaclopramide (10mg/kg p.o) respectively. 1 hr after treatments all the groups of animals were administered 4% activated charcoal meal and 20 min later killed by cervical dislocation for determination of intestinal transit. The small intestine was removed from the pyloric sphincter to the ilioceacal junction and the distance travelled by the charcoal meal was noted and expressed as percentage of intestinal transit using following formula13 .
Cholinergic system: Inhibitory effects of Chlorpromazine Chlorpromazine (5 mg/kg p.o.) produced significant (P < 0.05 Table 1) attenuation of intestinal transit by 28.85% when compared with vehicle treated group. In Chlorpromazinel-pretreated group, administration of benzene extract (400 and 800mg/kg) inhibits the delay of intestinal transit while the significant effect was observed at higher (800mg/kg p.o.) dose only (P < 0.05 Table 1). Chlorpromazine able to produce inhibition of intestinal transit upto 24.47 % in at lowest dose benzene extract of Piper betel.
DISCUSSION The gastrointestinal tract is in a continuous state of contraction, relaxation and secretion. These functions are controlled by neurohumoral systems, which in turn are regulated by various receptor systems, such as cholinergic, adrenergic, serotonergic, opioidergic and calcium channels4-11, 15.The results of the present study indicate that Piper betel accelerate the intestinal transit dose dependently. I found to accelerate the transit by 82.32 and 89.5% at dose of 400 and 800mg/kg of benzene extract of Piper betel respectively. The significant (P < 0.05 Table 1) acceleration was observed at higher dose (800mg/kg) compared to vehicle treated group (Table 1 and Figure 1). Various agents used to evaluate the pathways for acceleration or attenuation of intestinal transits e.g.
Chlorpromazine used for evaluation calcium channel10, 11 , clonidine used for adrenergic pathway14 , naloxone in opioidergic pathway9 , ondasetron in serotonergic system7 , and atropine in cholinergic mechanism4 . In present study we used chlorpromazine was used for delaying in the transit time by interfering calcium channel pathways11 . Calcium is involved in the initiation of contraction of smooth muscle10, 11. The visceral smooth muscle has a poorly developed sarcoplasmic reticulum and the increase in intracellular calcium concentration is primarily due to Ca2+ influx from the extracellular fluid via voltagegated Ca2+ channels15. The L-type calcium channel is present in many cells and it is the main source of Ca2+ for contraction of smooth muscle16. The result of the study found that chlorpromazine significantly inhibit the response of Piper betel by inhibiting the availability of calcium from extracellular sites represent from the result of inhibition of intestinal transit upto 24.47% compared to vehicle control (Table 2) indicating the involvement of Ca2+ channels in normal physiology of small intestinal motility (Table 2). When Chlorpromazine treated group was administered with Piper betel, it reverses the delay of intestinal transit induced by chlorpromazine (Figure 1).
This finding indicates that Piper betel possibly acts through calcium channel. In present study Piper betel increases the intestinal transit possibly by increasing the intracellular calcium concentration through calcium channel. Since, Chlorpromazine could able produce upto 24.47 % inhibitions of intestinal transit in presence lowest dose of Piper betel while in presence of higher dose chlorpromazine was able to inhibit by 15.85% only which was comparable to standard Metaclopramide (Table 1). This indicates Piper betel could partly produce acceleratory effect by increasing the intracellular calcium concentration through calcium channel and also by some other pathways as Chlorpromazine could not completely prevent the acceleratory effect of Piper betel.
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