Background Ficus nervosa (FN) leaves have been traditionally used in ethnomedicine for the treatment of various ailments, this study aimed to investigate the cold methanol extract of FN for its antioxidant, analgesic, antidiarrheal, and thrombolytic properties, along with preliminary phytochemical screening to identify key bioactive compounds and acute toxicity test to assess the safety profile of the extract. Methods In this study, we conducted an initial investigation to identify the major phytochemical groups present in the leaves of Ficus nervosa. Using conventional phytochemical screening methods on cold methanol extracts, we consistently identified phenols and flavonoids as the predominant bioactive compounds. Following this phytochemical characterization, we assessed the biological activities of the extracts across multiple models. The antioxidant capacity and free radical-scavenging ability of the leaves extracts were evaluated using the DPPH (2,2-Diphenyl-1-picrylhydrazyl) assay. Analgesic efficacy was assessed through acetic acid-induced writhing and electrically induced heat nociception tests. Antidiarrheal effects were examined using castor oil-induced diarrhea and an enteropooling assay in mice. To determine thrombolytic activity, we conducted a human blood clot lysis assay, while preclinical toxicity testing was conducted to evaluate the safety profile of the extracts. Statistical analysis was performed using one-way ANOVA with Duncan's Multiple Range Test (DMRT) to validate the significance of the results. Results The study of Antioxidant Potential, the ESF's (ethyl soluble fraction) antioxidant capacity, measured at 409.91 +/- 0.59 mg of ascorbic acid equivalent (AAE)/g in the phosphomolybdenum assay, signifies a strong free radical scavenging activity. This level of antioxidant action, alongside a potent free radical neutralizing concentration (IC?? of 9.99 +/- 0.41 ug/mL), suggests that the ESF could effectively counteract oxidative stress. Analgesic activity at 500 mg/kg, FN extract produced a 68.05% suppression of writhing in mice, which was statistically significant (p<0.001) compared to standard morphine's (74.05%) inhibition. The extract also significantly prolonged reaction latency to thermal-induced pain in hotplate model. The antidiarrheal activity showed 52.09% and 59.0% suppression of defecation, compared to standard loperamide's 73.1%, with statistically significant p-values (p<0.04 and p<0.01). These effects, combined with the notable decrease in intestinal fluid accumulation and defecation frequency in enteropooling assays, indicate that FN extract might interact with intestinal motility or fluid secretion pathways, providing antidiarrheal benefits. Additionally, the chloroform-soluble fraction of FN exhibited moderate thrombolytic activity, achieving 39.79% clot lysis compared to streptokinase (69.52%). Although this thrombolytic potential is moderate, it indicates that the fraction contains compounds with some fibrinolytic properties, which could be useful as adjunctive agents in preventing thrombosis. Conclusion Overall, the cold methanol extract of FN leaves demonstrates the therapeutic potential in preclinical settings. Future research is warranted to isolate the specific bioactive compounds and explain their mechanisms of action to further support the development of new treatments and contributing to modern medicinal practices based on this plant leaves.
