Black seed (Nigella sativa) extract enhances early and late apoptosis through activation of caspase-3 mediated regulatory pathway in LC540 cells: A network pharmacological and molecular docking approach.

Testicular cancer continues to rise in incidence globally, and conventional chemotherapy is often associated with severe side effects that significantly impact patient's quality of life. Identifying safer alternative therapies is becoming crucial day by day which leads to focus on naturally available phytocompounds with high bioactivity. Over the period of time, Nigella sativa (N. sativa) has garnered attention due to the presence of its rich bioactive compounds with antioxidant and anticancer properties, providing potential therapeutic benefits with minimal side effects. Since we used a network pharmacological based in-silico approach combined with in-vitro and in-vivo efficacy testing of N. sativa against testicular cancer. Molecular docking studies showed significant interactions between N. sativa phytochemicals and critical proteins involved in testicular and other cancer related pathways. Biochemical assays revealed decreased ROS levels with enhanced antioxidant enzyme activities such as SOD, CAT, GSH and reduced LDH levels. AO/PI staining further corroborated the enhanced apoptosis and necrosis rates in treated cells. m-RNA analysis demonstrated notable expression of inflammatory and apoptotic genes such as casp-3 and key testicular markers such as oct4, sox2 and other pro inflammatory cytokines. Histomorphological analysis of zebrafish testis showed decreased morphological alterations. With solid evidence of its anticancer effects through multiple biological mechanisms, including apoptosis induction, oxidative stress reduction, and oncogene suppression, these findings warrant further exploration of N. sativa as part of integrative cancer therapies to improve outcomes for patients with testicular cancer.