TITLE OF THE ARTICLE: EXOINTERACTIVE AMELIORATION AGAINST METHOTREXATE INTOXICATION IN MOUSE SPERMATOZOA

Abstract

Objective: To assess the synergistic potential using an exointeractive association between Murraya koenigii hydromethanolic leaf extract and the synthetic compound quercetin to offset methotrexate induced toxicity in mouse cauda epididymal spermatozoa. Methods: Healthy adult male albino mice were divided into eight groups (N= 6/group) and were treated with methotrexate intraperitoneally (10mg/kg body weight) at intervals of every 48 hours for 15 days, to combat this toxic effects of the drug, Murraya koenigii hydromethanolic leaf extract (250mg/kg body weight) and quercetin (15mg and 30mg) alone as well as in combination, were given by oral administration. Various sperm parameters such as sperm count, motility and viability followed by Hypo-osmotic Swelling (HOS) test, apoptosis test, Sperm Toroid Integrity (STI) and Sperm Chromatin Dispersion (SCD) assay. Statistical analysis used: Statistical analysis was carried out using Graphpad Prism Version 7 to verify levels of significance at the P < 0.001 value in one-way ANOVA. Results: Methotrexate induced a substantial reduction in sperm count, motility and viability which induces apoptotic changes, also resulted in disruption of sperm toroid protamine-DNA binding, increased fragmentation of nuclear DNA by decreased sperm chromatin dispersion and low percentage of swollen sperm representing membrane integrity loss. whereas Murraya koenigii hydromethanolic leaf extract and quercetin proved effective in reversing the drug induced toxicity, especially with the 15 mg dose of quercetin along with the plant extract brought about significant amelioration than 30mg dose of quercetin due to its pro-oxidant effects. Conclusions: To reduce the toxic manifestations of methotrexate, this exointeractive approach proved efficient.