Sequential Exposure to Multiple Antibiotics Depleted Glutathione in Hepatocytes, Disrupted Hematological Profile, and Inflammatory Responses in A Mouse Model

Abstract

Antibiotic misuse is considered a global threat due to drug-induced microbial resistance and disruption of vital biochemical and physiological processes. The aim of this study was to elucidate the impact of antibiotic-elicited dysbiosis on gut microbiota, oxidative stress, and inflammatory responses in a mouse model. Our experimental design simulated a poly-pharmacy situation where people are exposed to multiple antibiotic regimes over a prolonged period. Mice were allotted into six groups: group one was the control; group two was treated with amoxicillin 9.62 mg/kg; group three was treated with amoxicillin-clavulanic acid 17.53 mg/kg; group four azithromycin 15.38 mg/kg; group five received amoxicillin 9.62 mg/kg, amoxicillin-clavulanic acid 17.53 mg/kg, and azithromycin 15.38 mg/kg successively (sequential administration of the selected antibiotics); and group six received same treatment as group five but with probiotics being administered daily during the 7-day interval between each course of antibiotic treatment. The experiment lasted 49 days, during which samples were collected for analyses of gut microbiota, organ weights, hematological parameters, liver and kidney function tests, reduced glutathione (GSH) levels, cytokines (via ELISA), and histology. The results showed that antibiotic-induced dysbiosis altered body weight and caused amoxicillin-related splenomegaly. Successive exposure to antibiotics decreased RBCs, hematocrit, and hemoglobin levels but elevated WBCs, lymphocytes, and monocytes. These changes were restored upon co-administration of probiotics. Antibiotic-induced dysbiosis caused dyslipidemia. Furthermore, exposure to antibiotics led to a depletion of reduced GSH in the liver and gut, indicative of oxidative stress. The pro-inflammatory cytokines TNF-α and IFN-γ in serum were increased during concurrent administration of antibiotics, indicating inflammation. Liver function enzyme markers ALT and AST were elevated, signifying liver injury. All these effects were ameliorated in the group co-treated with probiotics. The findings from this study provide compelling evidence that exposure to multiple antibiotics over some time results in gut dysbiosis. This disrupts normal microbiota growth, affects the hematological profile, alters cytokine balance, increases oxidative stress, and impacts immune function