|Year : 2016 | Volume
| Issue : 4 | Page : 263-266
The effects of ocimum sanctum aqueous extract on intestinal motility of wistar albino rats by in vitro study
K Maheshkumar1, S Tamilmani2, R Sheeladevi2
1 Department of Physiology, Sri Ramachandra Medical College and Research Institute, Sri Ramachandra University, Chennai, Tamil Nadu, India
2 Department of Physiology, Dr. ALM PG IBMS Campus, University of Madras, Chennai, Tamil Nadu, India
|Date of Web Publication||15-Nov-2016|
Dr. K Maheshkumar
Department of Physiology, Sri Ramachandra Medical College and Research Institute, Sri Ramachandra University, Porur, Chennai, Tamil Nadu
Source of Support: None, Conflict of Interest: None
Introduction: Ocimum sanctum has been in use for thousands of years because of its diverse healing property in various conditions. Hence, to evaluate its effect on intestinal motility, the present study aimed to find the effects of O. sanctum on intestinal motility of Wistar albino rats by in vitro study. Materials and Methods: Aqueous extract of O. sanctum was prepared from mature leaves, and from this stock solution, various dilutions of 1:10,000, 1:1000, 1:100, and 1:10 were prepared and used. Thirty healthy adult Wistar-stained male albino rats were selected and divided into five groups (each group comprises six animals): control, 1:10,000, 1:1000, 1:100, and 1:10 dilution groups. Intestinal segments 2-3 cm below the duodenum was removed from the intestinal segment and mounted in the Dales Organ Bath with Tyrode's solution and respective contraction responses for the various dilutions were recorded separately in the physiograph. Results: O. sanctum aqueous extract showed that the amplitude and force of contractions in various dilutions have increased when compared to the control group in the intestinal motility. Among the various dilutions, 1:100 dilution group showed significant (P < 0.01) difference in the contraction than the other dilutions. Conclusion: The increase of amplitude with an increase in duration indicates that O. sanctum could slow down the intestinal motility. This reveals that the aqueous extract of O. sanctum contains pharmacologically active substances with antidiarrheal properties.
Keywords: Diarrhea, intestinal motility, medicinal properties, Ocimum sanctum
|How to cite this article:|
Maheshkumar K, Tamilmani S, Sheeladevi R. The effects of ocimum sanctum aqueous extract on intestinal motility of wistar albino rats by in vitro study. Int J Health Allied Sci 2016;5:263-6
|How to cite this URL:|
Maheshkumar K, Tamilmani S, Sheeladevi R. The effects of ocimum sanctum aqueous extract on intestinal motility of wistar albino rats by in vitro study. Int J Health Allied Sci [serial online] 2016 [cited 2023 Mar 28];5:263-6. Available from: https://www.ijhas.in/text.asp?2016/5/4/263/194130
| Introduction|| |
Ocimum sanctum commonly known as "Holy Basil" is a herbaceous plant found throughout the Southern Asian region. It is widely cultivated for food in many homes and temple gardens due to its religious significance. It has been in use for thousands of years in traditional medicine for its diverse healing properties. Ayurvedic practice recommends Tulsi in several formulations to enhance immunity and metabolic functions as well as in the management of respiratory problems.,, The leaf of O. sanctum has been reported to contain various bioactive phytochemicals such as saponin, flavonoids, triterpenoids, and tannins. Various other specific bioactive compounds isolated from O. sanctum were also studied for its pharmacological effects in the previous studies., The use of O. sanctum leaves in conditions including catarrhal bronchitis, bronchial asthma, dysentery, dyspepsia, skin diseases, chronic fever, hemorrhage, helminthiasis, and ringworms has been reported earlier., Fresh leaves of Basil taken together with black pepper are used as a prophylactic for malaria. Using the decoction of Tulsi leaves for cold is a popular remedy in the herbal medicine. In one study, it was observed the wound healing effect of aqueous extract of O. sanctum in rats. Linoleic acid present in different amount in the fixed oil of different species of O. sanctum has the capacity to block both the cyclooxygenase and lipoxygenase pathways of arachidonate metabolism and this could be responsible for the anti-inflammatory activity. Eugenol and the essential oils have also shown to be immune stimulant claiming the therapeutic potential of O. sanctum in immunological disorders associated with immune suppression.
No studies have been done so far to find the effects of O. sanctum on intestinal motility in healthy rats either in vivo or in vitro. Hence, the present study aimed to find the effects of O. sanctum on intestinal motility of Wistar albino rats by in vitro study.
| Materials and Methods|| |
Healthy adult Wistar-stained male albino rats, weighing 180-200 g (3 months old), were used in this study. The Institutional Animal Ethical Clearance (IAEC No: January 19, 2013) was obtained. The rats were kept in the animal room with controlled ambient temperature (24°C ± 2°C), humidity, and light. They were fed with standard rat feeds and water from the animal house department. Five groups of animals were used in this study and each group had six animals. The different groups used in this study are given in [Table 1]. The plant was collected at Chennai, Tamil Nadu. O. sanctum was identified and authentication (No.ARC/RR/2004/1091/September. 2004) was obtained from the experts. Mature greenish leaves of O. sanctum were collected and washed with sterile distilled water until the removal of all debris. These leaves were subjected to shade drying at room temperature for 2-3 weeks. One hundred grams of leaves was weighed and powdered. Briefly, aqueous extracts were prepared by soaking the O. sanctum powder in sterile distilled water, filtered through sterile cotton wool and 0.2 μ syringe filter, and the filtrate was freeze-dried and stored at – 20°C until use. For preparing stock solution, 2 mg of the aqueous extract was taken diluted in 9.4 ml of distilled water. From the stock, various dilutions of 1:10,000, 1:1000, 1:100, and 1:10 were prepared and used. Normal rats were sacrificed by cervical dislocation in the morning after overnight fasting. The abdomen was cut open and intestinal segments of 2.5-3 cm length below the duodenum were removed. The segments were flushed of luminal contents carefully with fresh Tyrode's solution and mounted vertically inside the central organ bath (15 ml capacity) containing Tyrode's solution, bubbled aerated (95% O2: 5% CO2 ) through hollow bent glass tube in Dales Organ Bath Apparatus (37°C, pH 7.4). The isolated organ was set up in such a way to record contractions from the longitudinal axis. Tension changes were recorded using isometric force transducers (Model T 305, INCO, India) to describe the changes in the contractile pattern of the isolated tissues, motility changes with and without drug application using student physiograph. Aqueous extract in various dilutions of O. sanctum was added to the bath and left in contact with the tissue for 30 s and then minimum of ten modified contractions were noted and washed out with normal saline. For each dilution, this procedure was repeated separately and the contractions were recorded.
All data were analyzed with the SPSS for Windows statistical package (version 19.0, SPSS Institute Inc., Cary, North Carolina, USA). Data were expressed as mean ± standard deviation (SD). Statistical significance between the different groups was determined by one-way analysis of variance. If the groups showed significant difference, Tukey's multiple comparison test was done. The significance level was fixed at P < 0.05.
| Results|| |
The segment of the intestine that submerged in the Tyrode's solution [Table 2] which has the same composition of extracellular fluid of rats showed rhythmic spontaneous contraction and relaxation and it represented control amplitude record. [Table 3] shows the amplitude (mm) and duration (second) of contraction of intestinal motility in controls and O. sanctum in various dilutions. The amplitude of intestinal contraction showed significant (P < 0.05) noticeable increase from control in 1:10,000, 1:1000, 1:100, and 1:10 dilution of O. Sanctum (df 4, F11). However, 1:100 dilution of O. Sanctum showed [Figure 1] marked increase than other dilutions of O. Sanctum and this increase, when compared to the same with other dilutions, was significant (P < 0.05). In the same way, duration of contraction in the presence of O. sanctum aqueous extract in various dilutions [Figure 2] showed marked increase in 1:100 dilution of O. Sanctum than other dilutions of O. Sanctum and this increase, when compared to the same with other dilutions, was significant (P < 0.05).
|Figure 1: Role of Ocimum Sanctum on Amplitude of contraction. Significance fixed at a level of P < 0.05. a: Control versus different dilutions. b: Represents significance of variation among the dilutions|
Click here to view
|Figure 2: Role of Ocimum Sanctum on duration of contraction. Significance fixed at a level of P < 0.05. a: Control versus different dilutions. b: Represents significance of variation among the dilutions|
Click here to view
|Table 3: Role of Ocimum sanctum on the amplitude (mm) and duration of contraction (s) |
Click here to view
| Discussion|| |
In the present study, we found that aqueous extract of O. sanctum in the lowest concentration has a significant role in altering the intestinal motility in the albino rats. The enteric nervous system is a collection of neurons in the gastrointestinal (GI) tract and constitutes the "brain in the gut" since it has the unique ability to control several GI functions, such as exocrine and endocrine secretions, motility, blood flow, and immune processes independent of the central nervous system., The sympathetic nervous system inhibits the intestinal movement and secretions, whereas the parasympathetic increases the motility and secretions. The important physiological role of local intestinal adrenergic mechanisms in reducing gut motility was well established, as discussed in several reviews.,, Gastric emptying is complex and reflects a variety of functions which include accommodation and coordinated relationship between the proximal/distal stomach and antro-pyloro-duodenal contractility. Several physiological conditions can be responsible for an altered GI transit time. When nerves or muscles in any portion of the digestive tract do not function with their normal strength and coordination, a person develops symptoms related to motility problems. Recently, medicinal plants have received more attention aiming its clinical application despite the concerns about their reliability and safety analysis. Animals have been used as experimental models for centuries and their uses have enabled researchers to make significant advances in many areas of human health and disease. The present study was designed based on this information. The observation indicated that O. sanctum have the action on intestinal motility. The increase of amplitude with an increase in duration indicated that O. sanctum could slow down the intestinal motility. An earlier study on O. sanctum in the form of aqueous extract diabetic rat has reported that it improves diabetes-induced delay in the intestinal motility by decreasing the lipid peroxidation because of the antioxidant activity. This variation of findings might be due to the difference in the study design because that was an in vivo study and delayed charcoal meal transit time in the intestine has been measured for the intestinal motility and this was totally different from the current study. Another study by Hannan et al. found that ethanol extract of O. sanctum decrease the gastric motility in the diabetic-induced rats which is in line with the results of the present study. In an in vivo study, the possibility of altering the activity of the intestine motility by other factors such as hormones and nerves could not be ruled out clearly. They have evaluated the intestinal motility in the form of gastric emptying and distal part of intestine transit time for the in vivo study. Since this is an in vitro study, we have used initial part of small intestine, especially 2-3 cm below duodenum to evaluate the motility. To the best of our knowledge, this was the first study showing the role of O. sanctum on intestinal motility in healthy albino rats and it confirms that aqueous extract of O. sanctum can alter the gut motility in this lowest concentration (0.02 mg/ml of aqueous extract).
| Conclusion|| |
This in vitro study shows that aqueous extract of O. sanctum in various dilution increases the amplitude and duration of the intestinal contraction. Therefore, it decreases the rate of intestinal contraction. This reveals that the aqueous extract of O. sanctum contains pharmacologically active substances with antidiarrheal properties. This property explains the basis for the effective use of the plant as an antidiarrheal agent in traditional medicine.
We would like to thank Dr. Ravindran, Professor and Head, Department of Physiology, Dr. ALM PG IBMS Campus, University of Madras, for his immense help and support in the study.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Das SK, Vasudevan D. Tulsi: The Indian holy power plant. Nat Prod Radiance 2006;5:279-83.
Mediratta PK, Dewan V, Bhattacharya SK, Gupta VS, Maiti PC, Sen P. Effect of Ocimum sanctum
Linn. on humoral immune responses. Indian J Med Res 1988;87:384-6.
Sakina MR, Dandiya PC, Hamdard ME, Hameed A. Preliminary psychopharmacological evaluation of Ocimum sanctum
leaf extract. J Ethnopharmacol 1990;28:143-50.
Singh S, Taneja M, Majumdar DK. Biological activities of Ocimum sanctum
L. fixed oil - An overview. Indian J Exp Biol 2007;45:403-12.
Jaggi RK, Madaan R, Singh B. Anticonvulsant potential of holy basil, Ocimum sanctum
Linn. and its cultures. Indian J Exp Biol 2003;41:1329-33.
Hakkim FL, Shankar CG, Girija S. Chemical composition and antioxidant property of holy basil (Ocimum sanctum
L.) leaves, stems, and inflorescence and their in vitro
callus cultures. J Agric Food Chem 2007;55:9109-17.
Kelm MA, Nair MG, Strasburg GM, DeWitt DL. Antioxidant and cyclooxygenase inhibitory phenolic compounds from Ocimum sanctum
Linn. Phytomedicine 2000;7:7-13.
Wagner H, Nörr H, Winterhoff H. Plant adaptogens. Phytomedicine 1994;1:63-76.
Warier PK. Eugenia jambolana
Linn. In: Warrier PK, Nambiar VP, Ramankutty C, editors. Indian Medicinal Plants. Madras, India: Orient Longman; 1995. p. 48-51.
Pandey BP. Anita. Economic Botany. Ramnagar, New Delhi: Chand and Company Ltd.; 1990. p. 294.
Shetty S, Udupa S, Udupa L, Somayaji N. Wound healing activity of Ocimum sanctum
Linn with supportive role of antioxidant enzymes. Indian J Physiol Pharmacol 2006;50:163-8.
Singh S. Comparative evaluation of antiinflammatory potential of fixed oil of different species of Ocimum
and its possible mechanism of action. Indian J Exp Biol 1998;36:1028-31.
Prakash P, Gupta N. Therapeutic uses of Ocimum sanctum
Linn (Tulsi) with a note on eugenol and its pharmacological actions: A short review. Indian J Physiol Pharmacol 2005;49:125-31.
Ganong W. Review of Medical Physiology. 22 nd
ed. Singapore: McGraw-Hill; 2005.
Epstein FH, Goyal RK, Hirano I. The enteric nervous system. N Engl J Med 1996;334:1106-15.
Bogers JJ, Timmermans JP, Scheuermann DW, Pelckmans PA, Mayer B, van Marck EA. Localization of nitric oxide synthase in enteric neurons of the porcine and human ileocaecal junction. Ann Anat 1994;176:131-5.
Daniel E. Pharmacology of adrenergic, cholinergic, and drugs acting on other receptors in gastrointestinal muscle.Mediators and Drugs I Gastrointestinal Motility II. BerlinHeidelberg: Springer; 1982. p. 249-322.
Bülbring E, Tomita T. Catecholamine action on smooth muscle. Pharmacol Rev 1987;39:49-96.
Szarka LA, Camilleri M. Methods for measurement of gastric motility. Am J Physiol Gastrointest Liver Physiol 2009;296:G461-75.
Di Lorenzo C, Youssef NN. Diagnosis and management of intestinal motility disorders. Semin Pediatr Surg 2010;19:50-8.
Baggio CH, Freitas CS, Rieck L, Marques MC. Gastroprotective effects of a crude extract of Baccharis
illinita DC in rats. Pharmacol Res 2003;47:93-8.
Robinson V. Less is more: Reducing the reliance on animal models for nausea and vomiting research. Br J Pharmacol 2009;157:863-4.
Kumar A. Ocimum sanctum
attenuates altered colonic contractility and intestinal transit in HFD-FED/STZ-treated type 2 diabetic rats. Res J Pharmacol Pharmacodyn 2012;4:341-5.
Hannan J, Ojo O, Ali L, Rokeya B, Khaleque J, Akhter M, et al
. Actions underlying antidiabetic effects of Ocimum sanctum
leaf extracts in animal models of type 1 and type 2 diabetes. Eur J Med Plants 2015;5:1.
[Figure 1], [Figure 2]
[Table 1], [Table 2], [Table 3]