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Nonsteroidal anti-inflammatory drugs have a number of side effects. However, some medicinal herbs have antiinflammatory properties with few side effects. This study aimed to determine the effect of the aqueous extract of Gongronema latifolium (AEGL) leaves on the level of tumor necrosis factor-α (TNF-α), transforming growth factor-β (TGF-β), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) in rabbits. Enzymelinked immunosorbent assay (ELISA) was performed to determine the effect of the AEGL on the activities of pro-inflammatory cytokine (TNFα), and regulatory cytokine (TGF-β) in rabbits. Also, a colorimetric method was performed to evaluate the levels of AST and ALT. Overall, the weight of the animals increased until the 21st day and then slightly decreased in the last week of treatment. AEGL showed significant (p< 0.05) inhibitory activity against pro-inflammatory and regulatory cytokines. The greatest inhibitory activities against TNF-α (21 days) and TGF-β (14 days) were obtained at a dose of 400 mg/kg of AEGL. No hepatotoxicity was observed within a 28-day period. This study showed that AEGL can be used as a safe herbal anti-inflammatory and immunomodulatory treatment. Further clinical trials are needed to approve this potential.