Individuals with lymphatic filariasis (LF) show elevated levels of pro-inflammatory cytokines and altered expressions of inflammatory genes, and are prone to secondary bacterial infection. Also markers of liver dysfunction were reported to be high with increased filarial infection. Acetaminophen (Paracetamol), an anti-pyretic drug was prescribed to LF individuals along with diethylcarbamazine (DEC) and albendazole (ALB) to alleviate pain and filarial fever within the therapeutic dosage. It is unclear, whether parasite-secreted toxins augment the action of acetaminophen and enhance liver dysfunction. Hence, an agent which can attenuate the acetaminophen toxicity during filarial infection is warranted. Trachyspermum ammi (T. ammi) is the richest source of thymol, reported to have the anti-filarial lead molecule. In the present study we evaluated the effects of thymol against acetaminophen-induced hepatotoxicity in in-vitro settings. In addition, we examined the synergistic effects of thymol against bacterial infection, free radicals and cytokine production. Our results reveal that acetaminophen (250 µg/ml) induces significant reduction in the viability of WRL-68 liver cells compared to cells without treatment. However, thymol at the same concentration restores the cell viability significantly (p = 0.031) by attenuating the toxicity within 24 h. Thymol inhibits the expression of Interleukin-6 (p = 0.043) and Interleukin-8 (p = 0.048) in WRL-68 cells and thereby provides maximum protection to liver cells from inflammatory insults. Calorimetric analysis shows the ability of thymol in scavenging hydrazyl (p = 0.004) and hydrogen peroxide(p = 0.008) free radicals efficiently. Thus thymol derived from T. ammi may be a good therapeutic agent in reducing the toxicity of acetaminophen, and may also aid in LF treatment and management in addition to the existing drugs.