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Studies on antidiarrhoeal activity of Jatropha curcus root extract in albino mice
Journal of Ethnopharmacology 70 (2000) 183 – 187 www.elsevier.com/locate/jethpharm
Short communication
Studies on antidiarrhoeal activity of Jatropha curcus root extract in albino mice A.M. Mujumdar *, A.S. Upadhye, A.V. Misar Agharkar Research Institute, Agarkar Road, Pune 411 004, India Received 8 March 1999; received in revised form 16 September 1999; accepted 2 October 1999
Abstract Use of Jatropha curcus L. roots in the treatment of diarrhoea is a common ethnobotanical practice in Konkan, a part of the Western coastal area of India. Roots of this species were undertaken for pharmacognostic studies and evaluation of antidiarrhoeal activity in albino mice. Successive solvent extraction was carried out using petroleum ether (60–80°C) and methanol. The methanol extract showed activity against castor oil induced diarrhoea and intraluminal accumulation of fluid. It also reduced gastrointestinal motility after charcoal meal administration in albino mice. The results indicate that action of J. curcus root methanol extract could be through a combination of inhibition of elevated prostaglandin biosynthesis and reduced propulsive movement of the small intestine. © 2000 Elsevier Science Ireland Ltd. All rights reserved. Keywords: Jatropha curcus roots; Antidiarrhosal activity; Albino mice; Castor oil; Charcoal meal
1. Introduction Jatropha curcus L. (Euphorbiaceae) is a native of tropical America. It is a soft-wooded, 5 – 50 foot high shrub, commonly grown as a fence near villages. Oil from Jatropha seeds has commercial value since it is used for the manufacture of candles, soaps, varnishes and lubricants. The oil is also used externally for the treatment of sciatica, * Corresponding author. Fax: +91-020-565-1542. E-mail address: [email protected] (A.M. Mujumdar)
dropsy, paralysis, rheumatism and certain skin diseases. A blue dye, obtained from the bark and leaves, is used for dying cloth and fishing nets (Anonymous, 1959; Agarwal, 1986; Ambasta, 1986). During an ethnobotanical survey of Raigad and Ratnagiri districts of Konkan area, a part of the West coast of India, use of J. curcus L. roots to control dysentery and diarrhoea was recorded. This use was cross-confirmed from several different localities and populations. In order to control these symptoms, a dose of two tablespoons of root suspension with butter milk is recommended,
0378-8741/00/$ - see front matter © 2000 Elsevier Science Ireland Ltd. All rights reserved. PII: S 0 3 7 8 - 8 7 4 1 ( 9 9 ) 0 0 1 6 7 - 1
184
A.M. Mujumdar et al. / Journal of Ethnopharmacology 70 (2000) 183–187
once or twice a day depending on the severity of the symptoms. It is interesting to note that this use is exactly opposite to the well-known purgative action of the seeds of this plant. It is reported that the root is triturated with a little asafoetida, and given with butter milk in dyspepsia and diarrhoea (Desai, 1975; Dymock et al., 1976; Nadkarni, 1976). Some of the pharmacognostic aspects of the roots have also been reported (Persinos et al., 1964; Raghunathan and Roma Mitra, 1982). In the present study, pharmacognosy of J. curcus roots and the antidiarrhoeal activity in albino mice were studied.
2. Materials and methods
2.1. Plant material Roots of J. curcus were collected from the Western ghat area of India in the winter of 1997–1998 in bulk quantities. A type specimen of the collected material has been deposited at Agharkar Herbarium of Maharashtra Association at Agharkar Research Institute, Pune, India (voucher specimen number, AHMA:17567).
2.2. Preliminary pharmacognosy and extraction procedure The roots of J. curcus were thoroughly washed with distilled water to remove dirt and soil, and were shade dried. Routine pharmacognostic studies including organoleptic tests, and macroscopic and microscopic observations were carried out to confirm the identity of the material. The roots were coarsely powdered and subjected to successive solvent extraction in soxhlet apparatus using petroleum ether (60 – 80°C) and methanol. Yields were 1.62 and 5.51%, respectively. These extracts were concentrated at reduced temperature and pressure on a rotary evaporator. The optimum conditions for experiments were decided on the basis of initial pilot experiments performed on three mice per treatment. J. curcus root extracts were administered at up to 1 g/kg
to an individual mouse in a group. There was no mortality due to this treatment. Hence, for further studies, 400 mg/kg (p.o.) of maximum dose was employed. In pilot studies, methanol extract showed antidiarrhosal activity, while the petroleum ether extract was devoid of any such activity. All further experiments were therefore carried out using methanol extract. The high-performance thin layer chromatography (HPTLC) studies of methanol extract were carried out on precoated silica gel plate (Merck 60 F 254) as the stationary phase and ethyl acetate:methanol:water (100:17:13) as the mobile phase. The extract was spotted using a Camag Linomat IV spotter. These plates were sprayed with methanolic sulphuric acid and were scanned on TLC scanner III using CAT software.
2.3. Experimental animals Swiss albino mice of either sex were originally obtained from the National Institute of Virology, Pune, India and have been maintained in the animal house facilities at the Agharkar Research Institute, Pune, for several generations for the past 14 years. They were housed in polypropylene cages in an air-conditioned area at 259 2°C with a 10:14 h light–dark cycle. They were given Amrut brand balanced animal feed and water ad libitum.
2.4. Castor oil induced diarrhoea Mice were divided into different groups as shown in Table 1 for the treatment either with the extract, vehicle or diphenoxylate HCl (5 mg/ kg). After 30 min, each of these animals was given 0.1 ml castor oil by oral route. The number of defecations per animal were recorded up to 4 h. The Jatropha extract and diphenoxylate HCl treated group exhibited a significant effect on castor oil induced diarrhoea, except at 50 mg/kg dose of the extract. For further studies, 100 mg/kg dose of extract was employed (Awouters et al., 1978).
T.H. Kang et al. / Journal of Ethnopharmacology 70 (2000) 177–182
the castor oil induced accumulation of intestinal fluid (Rao et al., 1997).
2.5. Small intestinal secretion Intestinal secretion was indirectly analysed by enteropooling assay. Groups of overnight fasted mice were treated with 100 mg/kg extract or vehicle orally, or chlorpromazine 30 mg/kg intraperitoneally 30 min before the oral administration of castor oil, 0.2 ml per mouse. These mice were sacrificed 30 min later, and the entire small intestine from each animal was weighed and their group average was calculated (Table 2). The difference in the weight of small intestine in control and castor oil treated groups was considered as Table 1 Effect of J. curcus root extract on castor oil induced diarrhoea in albino micea Group
Control
Number of mice
30
185
Mean number of defecations
2.6. Small intestinal transit The effect of the extract on small intestinal transit was studied in overnight fasted mice which were divided in different groups. These groups were control, vehicle, 100 mg/kg extract by oral treatment and 5 mg/kg atropine sulphate intramuscular route. Thirty minutes after the treatment, these mice were given 0.2 ml charcoal meal (3% charcoal in 5% gum acacia) by oral route. All animals were sacrificed after 20 min, the stomach and intestine removed, and the distance travelled by charcoal with reference to total length was calculated to express the percentage of distance travelled (Rao et al., 1997).
2.7. Drugs used
11.3090.54
Extract 50 mg/kg 100 mg/kg 200 mg/kg 400 mg/kg
10 10 10 10
9.29 0.42* 8.790.38* 6.890.34* 6.190.82*
Diphenoxylate HCl 5 mg/kg
10
4.39 0.74*
Data show mean 9 S.E.M. * Significant as compared with control PB0.05.
a
The extracts of J. curcus root were concentrated and suspended in 1% carboxy methyl cellulose (CMC) in water for administration. Castor oil was refined pure from Paras Chemical Industries. Chlorpromazine hydrochloride was purchased from Rhone Poulenc (India) Ltd. Charcoal was from E. Merck (India) Ltd. Atropine sulphate was from Loba Chemicals, and solvents were SQ grade of Qualigens fine chemicals.
2.8. Statistica1 analysis Table 2 Effect of J. curcus root extract on castor oil stimulated intraluminal fluid accumulation in the small intestine of mouse gut Experimental group
Weight of small in- Castor oil induced testine (mg/20 g 9 intraluminal fluid S.E.M.) (mg)
Control Castor oil CMC Extract (100 mg/kg) Chlorpromazine (30 mg/kg)
1062 980 1606 980* 1618 970 1342 9100 c
– 544 556 280 c
1083 990 c
21 c
* Significant as compared with control group, PB0.05. c Significant as compared with castor oil treated group, PB0.05.
All results were reported as mean9 S.E.M. These results were further analysed by using Student’s t-test to calculate significance of the results. P values less than 0.05 were considered significant.
3. Results J. curcus roots are tap roots with tuberous appearance and numerous branches. Young roots are soft. Fresh material is yellowish white, bitter with a typical agreeable aroma. The root shows longitudinal corrugations and well-developed lenticels. The transverse section of a mature root
186
T.H. Kang et al. / Journal of Ethnopharmacology 70 (2000) 177–182
Table 3 Effect of J. curcus root extract on gastrointestinal transit in mice Group
Number of mice used
Distance travelled by charcoal marker as % of total length of small intestine (mean 9 S.E.M.)
% Inhibition
Control Extract (100 mg/kg) Atropine sulphate (5 mg/kg)
10 10 10
83.38 94.75 67.0593.95* 39.359 6.86*
– 19.55 52.81
* Significant as compared with control, PB0.05.
shows well-developed lenticels, parenchymatous periderm and 16–25 layered tangentially elongated thin-walled cells. The phelloderm shows the presence of prismatic and flake-shaped calcium oxalate crystals singly or in groups. The zone of secondary phloem is comparatively broad. The secondary tissue shows a rosette type of calcium oxalate crystals and vessels are diffused. Three major peaks were observed on HPTLC at 366 nm with a 400 k filter at 0.10, 0.23 and 0.86 Rf. The densitometric proportion of these spots was 12.31, 51.44 and 36.26%, respectively.
3.1. Castor oil induced diarrhoea The methanol extract showed dose dependent inhibition of castor oil induced diarrhoea. This effect is significant as compared with control at 100, 200 and 400 mg/kg methanol extract and 5 mg/kg diphenoxylate, as shown in Table 1.
3.2. Small intestinal secretion The castor oil induced intraluminal accumulation of fluid is inhibited by 49.53% at 100 mg/kg extract. The reference drug chlorpromazine at a dose of 30 mg/kg reduced intestinal secretion by 96.14%. Both these values were significant as compared with control, as shown in Table 2.
3.3. Small intestinal transit The results of the present study revealed that the methanol extract at 100 mg/kg and atropine sulphate at 5 mg/kg significantly inhibited the gastrointestinal transit of charcoal in mice by
19.59 and 52.81%, respectively, as compared with control as shown Table 3.
4. Discussion In the present investigation, methanol extract of J. curcus roots has shown dose dependent antidiarrhoeal activity in a castor oil induced model in albino mice. This activity is significant at a dose of more than 100 mg/kg. Furthermore, this observation was also substantiated by significant action or castor oil induced intraluminal accumulation of fluid by indirect enteropooling assay in mice. The experiments carried out on gastrointestinal tract motility after charcoal meal administration have shown a reduction in the propulsive movement of small intestine after pre-treatment with methanol extract or atropine. These observations demonstrate the inhibitory effect of J. curcus root extract on castor oil induced diarrhoea, intraluminal fluid accumulation and peristaltic activity in small intestine. Prostaglandins contribute to the patho-physiological functions of the gastrointestinal tract, and also acts on the local electrical and mechanical activities of ileal circular muscles (Sanders, 1984). Castor oil increases peristaltic activity and produces permeability changes in the intestinal mucosal membrane to electrolytes and water (Bruton, 1985). Induction of diarrhoea by castor oil is through elevated prostaglandin biosynthesis (Awouters et al., 1978). From these observations, it can be deduced that the extract may act through inhibition of prostaglandin and reduction in propulsive movement of small intestinal tract. Present results are consistent with the
A.M. Mujumdar et al. / Journal of Ethnopharmacology 70 (2000) 183–187
reports on Nelumbo nucifera rhizome or plant constituents like bisnore (bisnordihydrotoxiferine) from Strychnos triner6is roots, ternatin from flowers of Egletes 6iscosa, flavonoids like apigenin, flavone, kaempferol, morin and rutin using the presented experimental models in animals, while investigating their mechanism of action (de Melo et al., 1988; Di Carlo et al., 1993; Mukherjee et al., 1995; Rao et al., 1997). Based on the present observations, the ethnobotanical use of J. curcus roots for antidiarrhosal activity can be explained.
Acknowledgements The authors are grateful to Dr A.D. Agate, Director, Agharkar Research Institute, Pune and Dr M.S. Kumbhojkar, In-charge, Botany Group for providing necessary facilities for this work.
References Agarwal, V.S., 1986. Economic Plants of India. Kailash Prakashan, Calcutta, p. 196. Ambasta, S.P. (Editor-in-Chief), 1986. The Useful Plants of India. Publication and Information Directorate, CSIR, New Delhi, p. 302. Anonymous, 1959. The wealth of India (Raw materials), Vol. V. Council of Scientific and Industrial Research Publication, New Delhi, p. 295. Awouters, F., Niemegeers, C.J.S., Lenaerts, F.M., Janssen, P.A.J., 1978. Delay of castor oil diarrhoea in rats: a new way to evaluate inhibitors of prostaglandin biosynthesis. Journal of Pharmacy and Pharmacology 30, 41–45.
.
187
Bruton, L.L., 1985. Agents affecting gastrointestinal water flux and motility, digestants; and bile acids. In: Gillman, A.G., Rall, T.W., Nies, A.S., Taylor, P. (Eds.), The Pharmacological Basis of Therapeutics Vol. 2, 8th ed. McGraw-Hill International editions, New York, p. 914. Desai, V.G., 1975. Aushadhi Sangraha. (Marathi). Shri Gajanan Book Depot, Dadar, Mumbai, p. 188. Di Carlo, G., Gautore, A., Izzo, A., Maiolino, P., Mascolo, N., Viola, P., Diurno, M.V., Capasso, F., 1993. Inhibition of intestinal motility and secretion by flavonoids, in mice and rats; structure – activity relationships. Journal of Pharmacy and Pharmacology 45, 1054. Dymock, C., Warden, C.J.H., Hooper, D., 1976 (Reprinted). Pharmacographica Indica — A History of the Principal drugs of Vegetable origin. Bishen Singh Mahindra Pal Singh. Dehra-Dun, p. 274. de Melo, M.F.F., Thomas, G., Mukherjee, R., 1988. Antidiarrhoeal activity of bisnordihydrotoxiferine isolated from the root bark of Strychnos triner6is (Vell.) Mart. (Loganiaceae). Journal of Pharmacy and Pharmacology 40, 79. Mukherjee, P.K., Das, J., Balasubramanian, R., Saha, K., Pal, M., Saha, B.P., 1995. Antidiarrhoeal evaluation of Nelumbo nucifera rhizome extract. Indian Journal of Pharmacology 22, 262. Nadkarni, A.K., 1976 (Revised). Dr. K.M. Nadkarni’s Indian Materia Medica Vol. I (originally K.M. Nadkarni (Ed.)). Popular Prakashan, Bombay, p. 705. Persinos, G.J., Quimby, M.W., Schermerhorn, J.W., 1964. A preliminary pharmacognostical study of ten Nigerian plants. Economic Botany 18, 329. Raghunathan, K., Roma Mitra (Eds.), 1982. Pharmacognosy of Indigenous Drugs Vol. I. Central Council for Research in Ayurveda and Siddha, New Delhi, p. 262. Rao, V.S.N., Santos, F.A., Sobreira, T.T., Souza, M.F., Melo, C.L., Silveira, E.R., 1997. Investigations on the gastroprotective and antidiarrhoeal properties of ternatin, a tetramethoxyflavone from Egletes 6iscosa. Planta Medica 63, 146. Sanders, K.M., 1984. Evidance that prostaglandins are local regulatory agents in canine ileal circular muscles. American Journal of Physiology 246, G361.
Short communication
Studies on antidiarrhoeal activity of Jatropha curcus root extract in albino mice A.M. Mujumdar *, A.S. Upadhye, A.V. Misar Agharkar Research Institute, Agarkar Road, Pune 411 004, India Received 8 March 1999; received in revised form 16 September 1999; accepted 2 October 1999
Abstract Use of Jatropha curcus L. roots in the treatment of diarrhoea is a common ethnobotanical practice in Konkan, a part of the Western coastal area of India. Roots of this species were undertaken for pharmacognostic studies and evaluation of antidiarrhoeal activity in albino mice. Successive solvent extraction was carried out using petroleum ether (60–80°C) and methanol. The methanol extract showed activity against castor oil induced diarrhoea and intraluminal accumulation of fluid. It also reduced gastrointestinal motility after charcoal meal administration in albino mice. The results indicate that action of J. curcus root methanol extract could be through a combination of inhibition of elevated prostaglandin biosynthesis and reduced propulsive movement of the small intestine. © 2000 Elsevier Science Ireland Ltd. All rights reserved. Keywords: Jatropha curcus roots; Antidiarrhosal activity; Albino mice; Castor oil; Charcoal meal
1. Introduction Jatropha curcus L. (Euphorbiaceae) is a native of tropical America. It is a soft-wooded, 5 – 50 foot high shrub, commonly grown as a fence near villages. Oil from Jatropha seeds has commercial value since it is used for the manufacture of candles, soaps, varnishes and lubricants. The oil is also used externally for the treatment of sciatica, * Corresponding author. Fax: +91-020-565-1542. E-mail address: [email protected] (A.M. Mujumdar)
dropsy, paralysis, rheumatism and certain skin diseases. A blue dye, obtained from the bark and leaves, is used for dying cloth and fishing nets (Anonymous, 1959; Agarwal, 1986; Ambasta, 1986). During an ethnobotanical survey of Raigad and Ratnagiri districts of Konkan area, a part of the West coast of India, use of J. curcus L. roots to control dysentery and diarrhoea was recorded. This use was cross-confirmed from several different localities and populations. In order to control these symptoms, a dose of two tablespoons of root suspension with butter milk is recommended,
0378-8741/00/$ - see front matter © 2000 Elsevier Science Ireland Ltd. All rights reserved. PII: S 0 3 7 8 - 8 7 4 1 ( 9 9 ) 0 0 1 6 7 - 1
184
A.M. Mujumdar et al. / Journal of Ethnopharmacology 70 (2000) 183–187
once or twice a day depending on the severity of the symptoms. It is interesting to note that this use is exactly opposite to the well-known purgative action of the seeds of this plant. It is reported that the root is triturated with a little asafoetida, and given with butter milk in dyspepsia and diarrhoea (Desai, 1975; Dymock et al., 1976; Nadkarni, 1976). Some of the pharmacognostic aspects of the roots have also been reported (Persinos et al., 1964; Raghunathan and Roma Mitra, 1982). In the present study, pharmacognosy of J. curcus roots and the antidiarrhoeal activity in albino mice were studied.
2. Materials and methods
2.1. Plant material Roots of J. curcus were collected from the Western ghat area of India in the winter of 1997–1998 in bulk quantities. A type specimen of the collected material has been deposited at Agharkar Herbarium of Maharashtra Association at Agharkar Research Institute, Pune, India (voucher specimen number, AHMA:17567).
2.2. Preliminary pharmacognosy and extraction procedure The roots of J. curcus were thoroughly washed with distilled water to remove dirt and soil, and were shade dried. Routine pharmacognostic studies including organoleptic tests, and macroscopic and microscopic observations were carried out to confirm the identity of the material. The roots were coarsely powdered and subjected to successive solvent extraction in soxhlet apparatus using petroleum ether (60 – 80°C) and methanol. Yields were 1.62 and 5.51%, respectively. These extracts were concentrated at reduced temperature and pressure on a rotary evaporator. The optimum conditions for experiments were decided on the basis of initial pilot experiments performed on three mice per treatment. J. curcus root extracts were administered at up to 1 g/kg
to an individual mouse in a group. There was no mortality due to this treatment. Hence, for further studies, 400 mg/kg (p.o.) of maximum dose was employed. In pilot studies, methanol extract showed antidiarrhosal activity, while the petroleum ether extract was devoid of any such activity. All further experiments were therefore carried out using methanol extract. The high-performance thin layer chromatography (HPTLC) studies of methanol extract were carried out on precoated silica gel plate (Merck 60 F 254) as the stationary phase and ethyl acetate:methanol:water (100:17:13) as the mobile phase. The extract was spotted using a Camag Linomat IV spotter. These plates were sprayed with methanolic sulphuric acid and were scanned on TLC scanner III using CAT software.
2.3. Experimental animals Swiss albino mice of either sex were originally obtained from the National Institute of Virology, Pune, India and have been maintained in the animal house facilities at the Agharkar Research Institute, Pune, for several generations for the past 14 years. They were housed in polypropylene cages in an air-conditioned area at 259 2°C with a 10:14 h light–dark cycle. They were given Amrut brand balanced animal feed and water ad libitum.
2.4. Castor oil induced diarrhoea Mice were divided into different groups as shown in Table 1 for the treatment either with the extract, vehicle or diphenoxylate HCl (5 mg/ kg). After 30 min, each of these animals was given 0.1 ml castor oil by oral route. The number of defecations per animal were recorded up to 4 h. The Jatropha extract and diphenoxylate HCl treated group exhibited a significant effect on castor oil induced diarrhoea, except at 50 mg/kg dose of the extract. For further studies, 100 mg/kg dose of extract was employed (Awouters et al., 1978).
T.H. Kang et al. / Journal of Ethnopharmacology 70 (2000) 177–182
the castor oil induced accumulation of intestinal fluid (Rao et al., 1997).
2.5. Small intestinal secretion Intestinal secretion was indirectly analysed by enteropooling assay. Groups of overnight fasted mice were treated with 100 mg/kg extract or vehicle orally, or chlorpromazine 30 mg/kg intraperitoneally 30 min before the oral administration of castor oil, 0.2 ml per mouse. These mice were sacrificed 30 min later, and the entire small intestine from each animal was weighed and their group average was calculated (Table 2). The difference in the weight of small intestine in control and castor oil treated groups was considered as Table 1 Effect of J. curcus root extract on castor oil induced diarrhoea in albino micea Group
Control
Number of mice
30
185
Mean number of defecations
2.6. Small intestinal transit The effect of the extract on small intestinal transit was studied in overnight fasted mice which were divided in different groups. These groups were control, vehicle, 100 mg/kg extract by oral treatment and 5 mg/kg atropine sulphate intramuscular route. Thirty minutes after the treatment, these mice were given 0.2 ml charcoal meal (3% charcoal in 5% gum acacia) by oral route. All animals were sacrificed after 20 min, the stomach and intestine removed, and the distance travelled by charcoal with reference to total length was calculated to express the percentage of distance travelled (Rao et al., 1997).
2.7. Drugs used
11.3090.54
Extract 50 mg/kg 100 mg/kg 200 mg/kg 400 mg/kg
10 10 10 10
9.29 0.42* 8.790.38* 6.890.34* 6.190.82*
Diphenoxylate HCl 5 mg/kg
10
4.39 0.74*
Data show mean 9 S.E.M. * Significant as compared with control PB0.05.
a
The extracts of J. curcus root were concentrated and suspended in 1% carboxy methyl cellulose (CMC) in water for administration. Castor oil was refined pure from Paras Chemical Industries. Chlorpromazine hydrochloride was purchased from Rhone Poulenc (India) Ltd. Charcoal was from E. Merck (India) Ltd. Atropine sulphate was from Loba Chemicals, and solvents were SQ grade of Qualigens fine chemicals.
2.8. Statistica1 analysis Table 2 Effect of J. curcus root extract on castor oil stimulated intraluminal fluid accumulation in the small intestine of mouse gut Experimental group
Weight of small in- Castor oil induced testine (mg/20 g 9 intraluminal fluid S.E.M.) (mg)
Control Castor oil CMC Extract (100 mg/kg) Chlorpromazine (30 mg/kg)
1062 980 1606 980* 1618 970 1342 9100 c
– 544 556 280 c
1083 990 c
21 c
* Significant as compared with control group, PB0.05. c Significant as compared with castor oil treated group, PB0.05.
All results were reported as mean9 S.E.M. These results were further analysed by using Student’s t-test to calculate significance of the results. P values less than 0.05 were considered significant.
3. Results J. curcus roots are tap roots with tuberous appearance and numerous branches. Young roots are soft. Fresh material is yellowish white, bitter with a typical agreeable aroma. The root shows longitudinal corrugations and well-developed lenticels. The transverse section of a mature root
186
T.H. Kang et al. / Journal of Ethnopharmacology 70 (2000) 177–182
Table 3 Effect of J. curcus root extract on gastrointestinal transit in mice Group
Number of mice used
Distance travelled by charcoal marker as % of total length of small intestine (mean 9 S.E.M.)
% Inhibition
Control Extract (100 mg/kg) Atropine sulphate (5 mg/kg)
10 10 10
83.38 94.75 67.0593.95* 39.359 6.86*
– 19.55 52.81
* Significant as compared with control, PB0.05.
shows well-developed lenticels, parenchymatous periderm and 16–25 layered tangentially elongated thin-walled cells. The phelloderm shows the presence of prismatic and flake-shaped calcium oxalate crystals singly or in groups. The zone of secondary phloem is comparatively broad. The secondary tissue shows a rosette type of calcium oxalate crystals and vessels are diffused. Three major peaks were observed on HPTLC at 366 nm with a 400 k filter at 0.10, 0.23 and 0.86 Rf. The densitometric proportion of these spots was 12.31, 51.44 and 36.26%, respectively.
3.1. Castor oil induced diarrhoea The methanol extract showed dose dependent inhibition of castor oil induced diarrhoea. This effect is significant as compared with control at 100, 200 and 400 mg/kg methanol extract and 5 mg/kg diphenoxylate, as shown in Table 1.
3.2. Small intestinal secretion The castor oil induced intraluminal accumulation of fluid is inhibited by 49.53% at 100 mg/kg extract. The reference drug chlorpromazine at a dose of 30 mg/kg reduced intestinal secretion by 96.14%. Both these values were significant as compared with control, as shown in Table 2.
3.3. Small intestinal transit The results of the present study revealed that the methanol extract at 100 mg/kg and atropine sulphate at 5 mg/kg significantly inhibited the gastrointestinal transit of charcoal in mice by
19.59 and 52.81%, respectively, as compared with control as shown Table 3.
4. Discussion In the present investigation, methanol extract of J. curcus roots has shown dose dependent antidiarrhoeal activity in a castor oil induced model in albino mice. This activity is significant at a dose of more than 100 mg/kg. Furthermore, this observation was also substantiated by significant action or castor oil induced intraluminal accumulation of fluid by indirect enteropooling assay in mice. The experiments carried out on gastrointestinal tract motility after charcoal meal administration have shown a reduction in the propulsive movement of small intestine after pre-treatment with methanol extract or atropine. These observations demonstrate the inhibitory effect of J. curcus root extract on castor oil induced diarrhoea, intraluminal fluid accumulation and peristaltic activity in small intestine. Prostaglandins contribute to the patho-physiological functions of the gastrointestinal tract, and also acts on the local electrical and mechanical activities of ileal circular muscles (Sanders, 1984). Castor oil increases peristaltic activity and produces permeability changes in the intestinal mucosal membrane to electrolytes and water (Bruton, 1985). Induction of diarrhoea by castor oil is through elevated prostaglandin biosynthesis (Awouters et al., 1978). From these observations, it can be deduced that the extract may act through inhibition of prostaglandin and reduction in propulsive movement of small intestinal tract. Present results are consistent with the
A.M. Mujumdar et al. / Journal of Ethnopharmacology 70 (2000) 183–187
reports on Nelumbo nucifera rhizome or plant constituents like bisnore (bisnordihydrotoxiferine) from Strychnos triner6is roots, ternatin from flowers of Egletes 6iscosa, flavonoids like apigenin, flavone, kaempferol, morin and rutin using the presented experimental models in animals, while investigating their mechanism of action (de Melo et al., 1988; Di Carlo et al., 1993; Mukherjee et al., 1995; Rao et al., 1997). Based on the present observations, the ethnobotanical use of J. curcus roots for antidiarrhosal activity can be explained.
Acknowledgements The authors are grateful to Dr A.D. Agate, Director, Agharkar Research Institute, Pune and Dr M.S. Kumbhojkar, In-charge, Botany Group for providing necessary facilities for this work.
References Agarwal, V.S., 1986. Economic Plants of India. Kailash Prakashan, Calcutta, p. 196. Ambasta, S.P. (Editor-in-Chief), 1986. The Useful Plants of India. Publication and Information Directorate, CSIR, New Delhi, p. 302. Anonymous, 1959. The wealth of India (Raw materials), Vol. V. Council of Scientific and Industrial Research Publication, New Delhi, p. 295. Awouters, F., Niemegeers, C.J.S., Lenaerts, F.M., Janssen, P.A.J., 1978. Delay of castor oil diarrhoea in rats: a new way to evaluate inhibitors of prostaglandin biosynthesis. Journal of Pharmacy and Pharmacology 30, 41–45.
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187
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