Delayed apoptosis upon the treatment of Hep-2 cells with black seed.

Nigella sativa (Black seed, BS) has been used to promote health and fight disease for centuries. The objectives of this investigation were: (1) to study whether agents such as cortisol and LPS alone or in combination induce cellular (Hep-2, laryngeal carcinoma) damage with time in culture (24, 48, and 72 hours) using apoptosis as a marker, (2) to determine if an immune stimulant such as BS, can protect Hep-2 cells from insult and ultimately thwart the programmed cells death mechanism. A total of 54 Hep-2 cell/tubes (50,000 cells per tube) were divided into six equal groups. Group one served as untreated control, while groups 2-6 were treated with either cortisol (10 ng/ml), LPS (10 micrograms/ml), BS (25 micrograms/ml), or a combination of LPS and cortisol and cortisol plus LPS plus BS, respectively. At the end of each phase the cells were harvested, heat fixed and stained with H&E to evaluate morphological changes. Immunohistochemistry, using antibodies against caspace-3 to evaluate cells undergoing apoptosis was conducted in all groups. The results of this study showed evidence of cells undergoing apoptosis at different magnitudes in all groups. However, the most dramatic change was seen in groups containing cortisol and LPS alone or in combination. This was supported by the fact that there were several adaptive responses observed in all phases. In addition, the exposure of BS to cells pretreated with cortisol and LPS showed evidence of protection against the progressive apoptosis.