Freeze dried extracts of Bidens biternata (Lour.) Merr. and Sheriff. show significant antidiarrheal activity in in-vivo models of diarrhea.
Diarrhea remains one of the main killers of children aged below five years. Traditional antidiarrheal remedies form a potentially viable source of novel low cost efficacious treatments in low resource settings. There is therefore a pressing need to scientifically evaluate these remedies.


OBJECTIVE
This study aimed to investigate the in vivo and in vitro antidiarrheal activity of freeze dried Bidens biternata, a herb used in traditional Ayurvedic medicine in the management of diarrhea.


METHODS
In the castor oil test, twenty (20) adult Sprague-Dawley rats were randomized to a negative control (normal saline, n=5), a positive control (5mg/kg loperamide, n=5), and two test groups. The low dose test group received 200mg/kg Bidens biternata extract (n=5) while the high dose test group received 400mg/kg B. biternata extract (n=5). Castor oil (4ml/kg) was then administered to the animals one hour after administration of the respective treatments after which the total mass of fecal output excreted after four (4) hours was determined. In the charcoal meal test fifteen (15) Sprague Dawley rats were randomized to a control group (normal saline 5ml/kg orally, n=5), a positive control group (atropine sulfate 0.1mg/kg i.p., n=5) and a test group (400mg/kg B. biternata extract, n=5). Charcoal meal was then administered via oral gavage to each rat thirty (30) minutes after the administration of the various treatments. The distance covered by the charcoal meal from the pylorus was then determined after sacrifice of the animals thirty minutes after the meal. In the enteropooling test twenty (20) Sprague-Dawley rats were randomized to a control group (5% v/v ethanol in normal saline, n=5), a positive control group (5mg/kg loperamide, n=5) and a test group (400mg/kg B. biternata extract, n=5). For each group prostaglandin E2 (PGE2) (100μg/kg) was administered immediately after the treatments. The animals were then sacrificed half an hour later and the volume of the small intestine contents determined. The effects of different concentrations of B. biternata extract (0.5. 1.0, 2.0, 3.0 and 5.0mg/ml) on jejunal contraction were investigated and a dose-response curve constructed using the experimental data after which The ED50 dose was determined. The effect of tamsulosin (α1 adrenergic blocker), yohimbine (α2 adrenergic blocker), propranolol (β adrenergic blocker) and naloxone (μ opioid blocker) on the contractile activity of the extract were also investigated. The experimental data were expressed as mean±standard error of mean (SEM) and then analyzed using one-way ANOVA followed by Tukey's post hoc test in cases of significance (set at p<0.05).


RESULTS
The freeze dried extracts of B. biternata had significant antidiarrheal effects in the castor oil induced diarrhea model (p<0.01) with the highest activity being observed at the 400mg/kg dosage level (1.66±0.81g vs. 4.54±0.51g control, p=0.01). B. biternata extract had significant effects on intestinal motility in the charcoal meal test compared to the control group (43.61±4.42% vs. 60.54±3.33%: p<0.05). B. biternata extract had a significant effect on PGE2 induced enteropooling (3.06±0.07ml vs. 4.74±0.10ml; p<0.001). The freeze dried extracts of B. biternata had a significant negative effect on the contractility of the isolated rabbit jejunum (p<0.001). The effects of the extract were significantly attenuated by tamsulosin (53.94±4.20% vs. 80.57±4.09%; p<0.01) and naloxone (53.94±4.20% vs. 73.89±7.26%; p<0.05). Yohimbine (p>0.05) and propranolol (p>0.05) however did not have any significant effect on the contractile activity of the extract.


CONCLUSIONS
The freeze dried extract of B. biternata possess significant antidiarrheal activity in both in vitro and in vivo models which appears to be mediated by modulating both the intestinal motility as well as the secretory activity. The results of this study also validate its traditional use as an antidiarrheal remedy.
Department of Medical Physiology, School of Medicine, University of Nairobi, P.O. Box 30197-00100, Nairobi, Kenya. Electronic address: gachkin@gmail.com.
Dennis Gacigi
DG
Kinuthia
Department of Medical Physiology, School of Medicine, University of Nairobi, P.O. Box 30197-00100, Nairobi, Kenya. Electronic address: awmuriithi09@gmail.com.
Anne W
AW
Muriithi
Department of Medical Physiology, School of Medicine, University of Nairobi, P.O. Box 30197-00100, Nairobi, Kenya. Electronic address: peterwaweru@uonbi.ac.ke.
Peter Waweru
PW
Mwangi
D018674:Adrenergic Antagonists / Q000494:pharmacology; D000818:Animals; D000930:Antidiarrheals / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D031176:Bidens / Q000737:chemistry*; D002368:Castor Oil; D003672:Defecation / Q000187:drug effects*; D003967:Diarrhea / Q000139:chemically induced / Q000188:drug therapy* / Q000378:metabolism / Q000503:physiopathology; D015232:Dinoprostone / Q000378:metabolism; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D005612:Freeze Drying; D005769:Gastrointestinal Motility / Q000187:drug effects*; D066298:In Vitro Techniques; D007419:Intestinal Secretions / Q000378:metabolism; D007583:Jejunum / Q000187:drug effects* / Q000378:metabolism / Q000503:physiopathology; D008139:Loperamide / Q000494:pharmacology; D008297:Male; D009292:Narcotic Antagonists / Q000494:pharmacology; D008517:Phytotherapy; D035261:Plant Components, Aerial; D010936:Plant Extracts / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D010946:Plants, Medicinal; D011817:Rabbits; D017207:Rats, Sprague-Dawley; D013997:Time Factors
D018674:Adrenergic Antagonists; D000930:Antidiarrheals; D009292:Narcotic Antagonists; D010936:Plant Extracts; D008139:Loperamide; D002368:Castor Oil; D015232:Dinoprostone
Adrenoceptor; Bidens biternata; Diarrhea; Jejunum; Motility; Opioid; Secretion