Studies on Toxicity and Anti – fertility of Cardiospermum halicacabum and Croton tiglium Seeds on Albino Rats

Abstract

Three experiments were carried out in this research work during 2008 – 2009 in order to investigate the potentiality of two medicinal plants, Cardiospermum halicacabum L. and Croton tiglium seeds, possessing toxic properties on albino rats, and to investigate for properties of Croton tiglium seed as an anti – fertility and anti – implantation on female albino rats. Preliminary phytochemical screening showed that both plants contained flavonoids and saponins but Croton seeds contain high alkaloids content while Cardiospermum halicacabum contains tannins and appreciable amount of cynoglycosides. No anthraquinone glycosides were detected in both plants. Albino rats of about 6 – 8 weeks age and of an average body weight between 120 – 180 grams and of both sexes were used for this study. After the acclimatization period, the animals were subjected to 3 experiments as follow: 1. Experiment I: Toxicity of Cardiospermum halicacabum on Albino Rats: Eighteen male albino rats of age 6 – 8 moths and 120 – 160 gms body weight were randomly divided into 3 groups. Group 1 served as control which given normal diet (300gms meat+ 700gms wheat, water corn oil and salt/ killo) purchased and given to the rats by the technicians of NCR,, while group 2 and group 3 were given daily diet contained 10% and 30% C. halicacabum powder respectively. Feeding continued daily for 4 weeks. No clinical signs were observed in the treated groups, and they look normal, without behavioral changes observed. No mortality recorded. A progressive body weight gain of the animals was observed in the three groups. The serum biochemical parameters showed no statistical differences in glucose concentration between control animals ( group 1) and animals in group 3, total protein and globulins also exhibited the same manner, no statistical changes from control value were seen, but remarkable significant reduction from control value was seen in albumin and urea levels at (P≤ 0.05 ). Lipids profile (total cholesterol and triglycerides), showed slight reduction in cholesterol value from control, but of no statistical significancy, in both treated groups of animals, but triglyceride showed remarkable significant decrease compared with control value. The result also showed that enzyme activities of AST (Aspartate Transferase) and ALP (alkaline phosphatase) showed no remarkable significant changes from control values in both groups of animals under treatment (group 2 and group 3), but the activity of the enzyme ALT (Alanine Transferase), showed remarkable rise activity. The haematological results showed no remarkable changes over control value. Histopathological examination of some animal’s body organs showed some changes occurred in small intestine and liver but of no clinical significancy. The mentioned changes were of minor alterations, but of no clinical significance. These results and findings may suggest that Cardiospermum halicacabum given as powder at concentrations of 10 % and 30 % mixed with the diet of the experimental albino rats for 28 days may had mild toxic effect on the animals these reflected in histopathological investigations on the intestine, liver and heart in which some histopathological alterations occurred and this also may not exclude the presence of toxic or lethal compounds in the plant. 2. Experiment II: Toxicity of Croton Seeds on Albino Rats: The second experiment which aimed to test the toxicity of Croton tiglium seeds on the experimental animals grouped in three groups treated as control group (group I) given normal animal diet, group II and group III given a 10 % and 20 % respectively of the Croton seeds mixed with animal diet for 2 weeks. The result revealed that animals in group I maintained normal body weight and looks normal throughout the 2 weeks of the experimental period, while animals in groups II and group III showed decrease in the body weight. Decrease in appetite, clinical signs such as watery diarrhea appeared few hours after the ingestion of the diet, no death occurred among the animals of treated groups who fed a diet containing 10% and 20 % of Croton tiglium seeds. The results also indicated that Croton seeds treatments brought about remarkable change on biochemical, haematological and histopathological parameters. 3. Experiment III: Anti - fertility and Anti - implantation Effect of Ethanol and Petroleum Ether Extract of Croton tiglium Seeds on Female Albino Rats: Croton seeds extracts were also tested. Petroleum ether Croton seeds extract administrated at a concentration of 200 mg/kg body weight daily for 7 days exhibited minor biochemical, haematological, and histopathological effects on the tested animals, but ethanol Croton seeds extract administrated at the same concentration resulted in profound alteration as mentioned in the treatment of animals with 20 % Croton seeds. As far as the antifertility and anti – implantation properties of the ethanol and petroleum ether extracts of the Croton seeds on the female animals, are concerned, the treated female animals were mated for 14 days with males after it subjected to treatment of petroleum ether seeds extract or ethanol seed extract, during which the animals were observed and signs of pregnancy were observed followed by examination of sexual and reproductive organs of the animals. The results revealed that female animals under petroleum ether extract treatment exhibited remarkable delay of pregnancy up to 3 months compared with control females who performed normal carriage and pregnancy and delivered at term, but the delivered litters although less in number compared with control females, and no complete absence of implantation in animal uterine horn occurred. But female rats of group II who treated with 200 mg/kg body weight daily for a week ethanol Croton seeds extract has showed 100% effective in preventing implantation sites in uterine horn of females animals. It has been postulated that endometruim of the animals uterus may be histopathologically affected resulted in decrease of contact and adhesion between blastocytes and uterine epithelium since histopathological examination of uterus indicated atrophy glands, odema and erosion of uterus superficial layers.