Methylprednisolone is a corticosteroid mainly used to treat severe allergies, inflammation, and flares of chronic illnesses such as the kidney. This corticosteroid functions by mimicking the effects of glucocorticoid hormones that are naturally secreted by our adrenal glands and consequently slows down an overactive immune system or replaces cortisol in the body. This steroid is prescribed in large doses primarily to control serious diseases involving the kidneys and brain. Most commonly, methylprednisolone is used to treat inflammatory manifestations of lupus nephritis- kidney disease caused by a systemic lupus erythematosus link. Medrol, a popular brand of methylprednisolone has solidified its role in autoimmune response control with the management of skin rashes and subsequent flare-ups in immunocompromised patients. However, despite the steroid’s known role as an anti-inflammatory, its effects on apoptotic signaling pathways within hypoxic conditions are poorly understood. AIM: This study aims to investigate the influence of Methylprednisolone on the apoptotic signaling pathway in Human embryonic kidney 293T (HEK293T) cells under hypoxic conditions, providing insight into potential implications for cell viability and survival. METHODS: The optimal doses of 0.5 mg and 0.25 mg were validated by crystal violet staining and subjected to hypoxic conditions. Apoptotic signaling pathways was analyzed using immunoblotting to examine the expression levels of key apoptotic markers. Additionally, gene expression levels for Bcl-2 were determined by RTqPCR. RESULTS: Methylprednisolone treatment modulates apoptotic signaling pathways in HEK293T cells under hypoxic conditions. Specifically, methylprednisolone inhibits apoptotic pathways, leading to enhanced cell survival and decreased apoptosis. These effects may be mediated through the regulation of pro-apoptotic and anti-apoptotic proteins, as well as downstream signaling cascades involved in cell death regulation. CONCLUSION: Methylprednisolone will inhibit apoptotic pathways in HEK293T cell lines under hypoxic conditions due to the Bcl-2 gene’s anti-apoptotic properties and Bax will be downregulated in response to the upregulation of Bcl-2 expression. By inhibiting apoptosis and promoting cell survival, methylprednisolone demonstrates potential as a therapeutic agent for mitigating tissue damage in conditions characterized by hypoxia or ischemia. Further research into the precise mechanisms underlying its effects on apoptotic pathways could provide valuable insights for the development of specified treatments for a range of inflammatory and ischemic diseases.