Effect of argemone oil and argemone alkaloid, sanguinarine on Sertoli–germ cell coculture

Several incidences of reduction in the fertility (sperm count) have been reported in India and worldwide as well. Adulteration of food and consumption of adulterated mustard oil with argemone oil (AO) are presumed to be the factors for reduction in sperm count. In the present study we have studied the exfoliation of germ cells from Sertoli cell, its viability after detachment, cytotoxicity and execution of apoptosis via mitochondrial pathway for different concentration of AO, argemone alkaloid (AA) and its major constituent sanguinarine (SA). A dose dependent increase in germ cell detachment and decrease in viability of detached germ cells were observed ( P < 0.05). A significant inhibition was observed via 3-(4,5-dimethylthiazol-2-yl)-2,5-dipehyl tetrazolium bromide (MTT) assay in the proliferative activity of germ cell and leakage of cytosolic enzyme was observed via Lactate dehydrogenase(LDH) assay ( P < 0.05). A time and dose dependent inhibition of mitochondrial membrane potential was observed ( P < 0.05). Treatment of Sertoli–germ cells with the lowest concentration of AO/AA and SA for 24 h resulted in 5.2- , 4.4- and 3.6-fold increase in the percentage of early apoptotic cells, respectively. This increase was enhanced to 8.3, 4.75 and 5.81-fold, respectively at 48 h in detached germ cells undergoing early apoptosis. These results suggest that alterations in germ cell apoptosis by a disruption in contact mediated communication between the Sertoli cells and germ cells, may subsequently lead to testicular impairment.