Aqueous extract of Azadirachta indica (neem) leaf induces generation of reactive oxygen species and mitochondria-mediated apoptosis in rat oocytes.

ABSTRACT

OBJECTIVE: Present study was aimed to determine whether aqueous neem leaf extract (NLE) induces generation of reactive oxygen species (ROS) and apoptosis through mitochondria-mediated pathway in rat oocytes. DESIGN: A controlled prospective study. SETTING: Laboratory research setting at Department of Zoology of Banaras Hindu University. ANIMAL(S) Forty eight sexually immature female rats that were 20-30 days of age. INTERVENTION(S) Sexually immature female rats were fed palatable dose of NLE (10 mg/g dry feed palate) for 10 days and then subjected to superovulation induction protocol. Thereafter, rats were euthanized, ovulated cumulus oocyte complexes were collected from oviduct and oocytes were denuded. MAIN OUTCOME MEASURE(S) Rate of morphological apoptotic changes, measurement of hydrogen peroxide, nitric oxide and cytochrome c concentrations, caspase-9, caspases-3 activities and DNA fragmentation in oocytes. RESULTS: In vivo NLE treatment induced morphological apoptotic changes were associated with increased hydrogen peroxide, nitric oxide and cytochrome c concentrations, caspase-9, caspase-3 activities and DNA fragmentation in oocyte. CONCLUSION: NLE induces generation of ROS that leads to oocytes apoptosis through mitochondria-mediated pathway.