Abortive and Teratogenicity of Ethyl Acetate Extract Fraction of Eugenia Jambolana Seed in Pregnant Rats

Introduction

Population control is of immense importance for individual and national welfare. Hence the fertility control is the most important and urgent mainstay of all biomedical and biosocial problems. Although a variety of synthetic contraceptive agents are available, these are not without side effects [1]. In recent years there has been considerable interest in plants with potential contraceptive properties. Research on Indian medicinal plants with contraceptive property has been exhaustively reviewed by so many researchers and scientists, but so far no single plant is available, which can safely use to prevent pregnancy and reproductive disorders [2].

Throughout the history women have tried to control or enhance their fertility with various levels of societal support. Many herbal remedies are traditionally used as contraceptive (to prevent the ovulation or fertilization), abortifacient (to prevent implantation) and emmenagogues (to prevent uterine flow) or Oxytosic (to stimulate uterine contraction particularly to promote labour) [3]. Many such plants are recommended in Ayurvedic, Unani and Folk medicines [4, 5, 6]. A good number of research works is going on medicinal plants for antifertility effect. In the same direction, present approach is being pursued to identify antifertility agent from the seeds of E. Jambolana. In our previous findings on the ethyl acetate extract of E. Jambolana seeds demonstrated their anti-implantation and antiestrogenic [7], abortifacient activity [8] in rats and mice. In the present study we have undertaken the investigation of post coital contraceptive activity and Teratogenicity property of chromatographic fractions of crude ethyl acetate extract of E. Jambolana seeds to elucidate its active ingredients.

Study of Developmental Toxicity and Teratogenicity

The fetuses were removed by opening the uterus and were placed in a sequential manner in 0.9% saline solution. All dead and live fetuses were counted and the following observations were made.

Statistical Analysis

One way analysis of variance [ANOVA] followed by Duncan’s multiple test were used to find out significant difference among mean values of each parameter of different experimental groups by fixing minimum significance level at P<0.05. Values with same superscript letters are not significantly [P<0.05] different whereas those with different superscript letters are significantly [P<0.05] different when compared to control.

Result

All values are expressed as Mean ± Standard error. The data was analyzed by one way ANOVA, Analysis of Variance. Values with same superscript letters are not significantly [P<0.05] different whereas those with different superscript letters are significantly [P<0.05] different as judged by Duncan’s Multiple Test (Figure 1). The study of developmental toxicity and Teratogenicity is given in Table 2. The litter size, body weight and body length of litters born in control were 5.94±0.32, 5.00±0.39 and 4.01±0.42cm respectively. There were no statistical significant differences between groups in the litter size, body weight and body length of litters throughout the post partum period. There were no noticeable visceral malformations observed in the offspring of control as well as in all treated rats. External examination of all fetuses revealed no abnormality between the groups when compared to controls. There were no noticeable skeletal and teratogenic malformations in the skull, vertebral column, Sterne brae, forelimbs, hind limbs, fore and hind paws were observed in the fetuses of control as well as in all treated rats.

Click to enlarge

Table 2: Effect of Fractions of ethyl acetate extract of E. Jambolana seeds on litters. All values are expressed as Mean ± Standard error. The data was analyzed by one way ANOVA, Analysis of Variance. Values with same superscript letters are not significantly [P<0.05] different whereas those with different superscript letters are significantly [P<0.05] different as judged by Duncan’s Multiple Test.

Discussion

In the present study, the chromatographic fractions of ethyl acetate extract of E. Jambolana seeds were tested for their abortifacient and Teratogenicity properties in female albino rat. The fraction - I was found to be most active fraction of ethyl acetate extract. The number of litters born as a result of this fraction I treatment was significantly less (nil) in comparison to control. This proves the pregnancy interceptive activity of fraction I of ethyl acetate extract of E. Jambolana seed.

There is much evidence that shows that some herbal drugs can be used as an abortifacient in mice, rats, rabbits and even humans [12, 13]. An agent that can disrupt pregnancy could be of obvious interest in human fertility control [14]. The implantation index, resorption index and pre-implantation loss are useful indices for evaluating the number of blastocycts implanted in the uterus and the underdeveloped [15]. Therefore, the increase in the resorption index by the fraction extract is an indication of failure in the development of the embryo. Such occurrence of fetal resorption suggests that interruption of pregnancy occurred after implantation of the foetus [16]. All these are indications of the pregnancy termination potential of fraction I of ethyl acetate extract of E. Jambolana seeds. Results of present study are in conformation with results obtained with Citrus hystrix [17]. Ethanolic extract of citrus hystrix in a dose of 1 gm/kg resulted in abortifacient effect in female albino rats. These findings are in agreement with those of Yakubu [18] who reported abortifacient activity of Senna alata leaves. Shibashi [19] reported Methanolic extract administration of Achyranthus aspera leaves caused abortifacient activity in pregnant rats. The resorption index and post implantation loss establishes correlation between the number of implanted blastocysts and those that have not developed [20, 21]. Similar observations were in agreements with those of Dabhadkar and Varsha [22] using Plumeria rubra pod extract in rats. The results of the present study also revealed that the fraction I extract was relatively non- embryotoxic as judged by the data on litter body size and the absence of any observable treatment related morphologic defects and skeletal and visceral malformation in the fetuses when compare to control. These findings are in agreement with Gebrie [23] who reported that the methanolic extract of the Rumex steudelli root did not show teratogenic effect in rats. Similar observations have been made with the findings of Chukwuka [24] and Abdulazeez [25] on Spondias and Carrica respectively.

Conclusion

The present study suggests that the chromatographic fraction I of ethyl acetate extract of Eugenia Jambolana possesses abortifacient activity and these findings could explain its traditional use as an abortifacient agent. Preliminary phytochemical analysis and TLC revealed that fraction I contains more of flavonoids and a mixture of compounds. The activity of the fraction I owes due to synergistic action of multiple metabolites. Therefore, it is hoped that this fraction can be further purified to get a potent compound and may be brought out as an effect contraceptive in near future.