Alpha-ketoglutarate supplementation ameliorates castor oil-induced diarrhea in mice: in silico and in vivo insights.

Alpha-ketoglutarate (α-kg) is an intermediate of the Krebs cycle and a precursor of glutamine and glutamate. Furthermore, α-kg contributes to the maintenance of intestinal barrier integrity. Diarrhea, a global health problem, is characterized by an increased frequency and liquidity of bowel movements and is closely associated with impaired intestinal function. As oral rehydration is the primary treatment modality available, the present study evaluated the antidiarrheal potential of α-kg in castor oil (CO)-induced diarrhea model and investigated its molecular interactions through in silico modeling. Molecular docking was performed to assess the binding affinity of α-kg with CFTR (3SI7), CaCC (5OYB), and Na+/K+-ATPase (3B8E). For the in vivo experiments, mice were divided into the following groups: vehicle, α-kg (120, 60, and 30 mg/kg), CO (10 mL/kg), and loperamide (5 mg/kg). α-kg was administered orally for five consecutive days. On the fifth day, one hour after the final dose, diarrhea was induced with CO. The loperamide group was pre-treated with loperamide prior to CO administration, whereas the CO group received only CO, and the vehicle group received distilled water. After four hours, the animals were euthanized, and the intestinal samples were collected to evaluate intestinal content, chloride ion levels, and oxidative stress markers (malondialdehyde, glutathione, and nitrite). α-kg reduced fecal outputs, intestinal fluid secretion, and oxidative stress markers, while enhancing Na+/K+-ATPase activity, preserving glutathione levels, and improving histomorphometric parameters and mucin content. In conclusion, α-kg exhibited favorable binding interactions with key molecular, antidiarrheal, and antioxidant effects, and restored intestinal histological parameters.