Contraception potential of neem oil: effect on pregnancy success in the mouse.

Abstract

The aim of this study was to find out the role and mechanism of action of neem oil as a postcoital fertility blocker in mouse. Female mice were injected with neem oil (20 or 40 microliters) surgically into each uterine horn on day 2 postcoitum (pc). Both the uterine horns of each mouse were injected. Arachis oil served as vehicle control. Pregnancy success was determined by the number of implanted embryos on day 8 pc and the number of live fetuses in the uteri on day 18 pc. Transforming growth factor-alpha (TGF alpha), epidermal growth factor (EGF), and epidermal growth factor receptor (EGFR) were immunolocalized in the paraffin-embedded sections of the uteri at 0600 hr on day 5 pc. The unimplanted embryos were assessed in the uteri at 2000 hr on day 5 pc. Uterine secretions were assessed for the leukocytes infiltration on day 4 through day 8 pc. The number of implantation sites on day 8 pc and the number of live fetuses on day 18 pc were lower in the neem oil-treated animals compared to their respective control animals at both the concentrations of neem oil (20 and 40 microliters/uterine horn). Neem oil also caused resorption of some embryos between day 8 pc and day 18 pc. In neem oil-treated mice, EGFR immunostaining decreased in the luminal and glandular epithelium and increased in the stroma as determined at 0600 hr on day 5 pc. Uterine secretions on day 4 through day 6 pc from the neem oil-treated mice showed massive leukocyte infiltration. Unimplanted preimplantation embryos, underdeveloped, degenerated, or at blastocyst stage, were recovered from the uteri after flushing at 2000 hr on day 5 pc from the neem oil-treated animals. A number of retrieved unimplanted embryos showed the direct attachment of the leukocytes to their zona pellucida. It is believed that the secretions of these leukocytes might be responsible for the underdevelopment of the early embryos and hence inhibition of implantation. The exact interaction of these leukocytes and their secretions with the early embryos is under investigation. Postcoital intrauterine treatment of neem oil during preimplantation period causes fertility block in mouse by lowering the EGFR localization in the luminal and glandular epithelium, by causing massive leukocytes infiltration into the uteri, by degenerating the early embryos, and by causing the postimplantation embryonic resorptions in the uteri. The possible mechanism of action of neem oil is discussed.