Gut microbial imbalances caused by unrestricted antibiotic use and exposure to chemical carcinogens contribute to liver and kidney toxicity: insights from in silico and in vivo studies

The gut microbiota is a complex community of microorganisms that play a vital role in human health; however, antibiotic administration can disturb this balance. Such disturbances have been linked to an increased risk of various diseases, including cancer. Notably, contamination of food with Aflatoxin B ₁ (AFB ₁ ) and Diethyl nitrosamine (DEN) has been shown to significantly raise the risk of liver cancer, as both agents are potent hepatocarcinogens associated with high incidences of hepatocellular carcinoma. This study examines the impact of widespread access to ampicillin (AMP) and ciprofloxacin (CPX)—two commonly used antibiotics in Nigeria—on the composition of the gut microbiota, dietary exposure to AFB ₁ and DEN, and subsequent liver and kidney toxicity in rats. Using network toxicology, this research also explores how CPX and AMP influence gut bacteria and how AFB ₁ and DEN trigger hepatorenal toxicity through redox and protective pathways. The study compares computational predictions with laboratory findings, identifying STAT3, EGFR, MAPK8, IKBKB, MMP2, MET, and NOS2 as key genes involved in these processes. Molecular docking results indicate that AFB ₁ , CPX, and AMP each bind strongly to EGFR (with affinities of − 8. 5, − 7. 7.8, and − 7. 7.6 kcal/mol, respectively), suggesting a potential combined effect on EGFR signalling. These interactions may provide insight into how changes in the gut microbiota contribute to toxicity involving both STAT3 and EGFR. In vivo validation was carried out using male Wistar rats (n = 90, 200g ± 20), divided into nine groups: Group 1 —control (2 mL saline, per os ); Group 2 —AMP (20 mg/kg, per os ); Group 3 —CPX (12. 5 mg/kg, per os ); Group 4 —DEN (200 mg/kg, i. p. ); Group 5 —AFB ₁ (2 mg/kg); Group 6 —AMP + DEN; Group 7 —CPX + DEN; Group 8 —AMP + AFB ₁ ; Group 9 —CPX + AFB ₁ . Two rats from each group were sampled before and after treatment to assess gut microbiota. The findings revealed that DEN, AFB ₁ , and combinations with AMP or CPX reduced body, liver, and kidney weights ( p < 0. 05 ). Co- treatments elevated serum transaminase, creatinine, and urea levels, while antioxidant enzyme activity, GSH, and TSH decreased ( p < 0. 05 ). Markers of inflammation, lipid peroxidation, alpha- fetoprotein, and caspase- 3 increased, whereas IL- 10 decreased in the liver and kidney ( p < 0. 05 ). Overall, AMP and CPX exacerbated gut dysbiosis and worsened hepatorenal toxicity with AFB ₁ and DEN, heightening the risk of carcinogenesis.