Exploring unrestricted antibiotic use gut microbiome imbalance and chemical carcinogens in liver and kidneys toxicity through computer modeling and in vivo study

The gut microbiota, vital for human health, can be disrupted by antibiotics, which has been linked to diseases such as cancer. Contamination of food with Aflatoxin B 1 (AFB 1 ) and Diethylnitrosamine (DEN), both strong liver carcinogens, increases liver cancer risk. This study examines how unrestricted use of common antibiotics (Ampicillin and Ciprofloxacin) affects gut microbes, combined dietary exposure to AFB 1 and DEN, and the development of liver and kidney toxicity in rats. Molecular Docking (MD) and Network Toxicology were used to investigate how AFB 1 , DEN, AMP, and CPX cause hepatorenal toxicity by interacting with key hub proteins involved in redox and protective cellular pathways. Ligand orientation and binding energies for STAT3 and EGFR were assessed using molecular docking. Gut microbial analysis shows that AMP and CPX induce antibiotic-driven dysbiosis, suppressing beneficial gram-positive bacteria like Lactobacillus and promoting the growth of opportunistic gram-negative bacteria such as Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa. These changes result in toxic metabolite production affecting the hepatic immune system. In the presence of carcinogens, AMP and CPX synergy may contribute to carcinogenesis in co-treated groups. STAT3, EGFR, MAPK8, IKBKB, MMP2, MET, and NOS2 were identified as important hub genes. AFB 1 (-8.5 kcal/mol), CPX (-7.8 kcal/mol), and AMP (-7.6 kcal/mol) showed high affinity for EGFR, indicating a possible synergistic role in EGFR signalling modulation. In vivo studies in male Wistar rats showed that co-treatment reduced body, liver, and kidney weights, raised serum hepatic transaminases, creatinine, and urea, decreased antioxidant enzymes, and increased inflammation markers, lipid peroxidation, alpha-fetoprotein, and caspase-3 activity, alongside reduced IL-10 in the liver and kidney. The data indicate that AMP and CPX cause gut dysbiosis and, together with AFB 1 and DEN, have a synergistic hepatorenal toxic effect that may be relevant in the onset of carcinogenesis.