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Summary
Methoxychlor, an organo-chlorine insecticide, has been found to be a potential female reproductive toxicant enhancing apoptosis mediated follicular atresia. It is suggested that induced geno-toxicity or DNA damagetriggers apoptosis;hence, the present study aimed at elucidating the potentials of Methoxychlor as a geno-toxicant, inducing granulosa cells apoptosis during the in vitro treatment. The granulosa cells of healthy antral follicles were exposed to different concentrations of MXC (1, 10 and 100 µg/ml) and antioxidant, N-Acetyl-L-cysteine (1, 5 and 10 mM) for 24, 48 and 72 hr exposure duration, following which the cells were subjected to terminal deoxynucleotidyltransferasedUTP nick end labelling and single cell gel electrophoresis (comet) assay.
MXC exposure enhanced DNA Fragmentation, as revealed by the increased incidence of dark brown condensed TUNEL positive cells in contrast with lightly brown TUNEL negative cells with maximum TUNEL positive cells were observed in 100 µg/ml MXC treated groups. Quantitatively, maximum geno-toxicity was exhibited at highest MXC treatment with percenttail DNA as 17.87 ± 0.85, 41.16 ± 3.94, and 47.73 ± 3.71 in comparison with control (0.65 ± 0.03, 2.91 ± 0.27, and 7.16 ± 1.39) after 24, 48 and 72 hr exposure duration, respectively. MXC treated groups exhibited Type 1-Type 3 comets with no Type 4 comets were observed as compared toType 0 comets in control groups. Supplementation of NAC led to significant (p < 0.05) decline in geno-toxicity in MXC treated groups with maximum amelioration observed at 5 and 10 mM. Similar trend was observed with increasing the exposure duration to 48 hr and 72 hr.
Consequently, increased DNA damage attributed to the granulosa cells apoptosis in response to Methoxychlor exposure resulted in follicular atresia associated infertility that was significantly combated by NAC supplementation, preventing the geno-toxicity induced cyto-toxicity in GCs.
