Study on the expression changes of hepatic Gluconeogenesis-related proteins induced by insulin Overdose-related liver injury.

PURPOSE: Insulin overdose severely disrupts hepatic glucose metabolism, leading to toxic changes in the liver that may play an important role in the diagnosis and identification of insulin overdose. METHODS: A rat model of insulin overdose was established in this study. ALT and AST levels were measured to assess liver function. Hematoxylin and eosin, oil red O, and periodic acid-Schiff staining were used to evaluate pathological liver injury. Ultrastructural changes in hepatocyte organelles were observed under a transmission electron microscope. The expression levels of liver gluconeogenesis-related proteins, including forkhead box protein O1 (FOXO1) and phosphorylated FOXO1 (p-FOXO1), phosphoenolpyruvate carboxykinase (PEPCK), glucose-6-phosphatase catalytic subunit (G6PC), chromosome region maintenance 1 (CRM1), and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein (14-3-3), were detected in the rat model and human autopsy liver tissues using immunohistochemistry and immunofluorescence. RESULTS: The insulin overdose group showed elevated ALT/AST, hepatocyte fatty degeneration, glycogen depletion, mitochondrial swelling, and cristae loss. The recovery group exhibited milder fatty changes, glycogen accumulation, and restored mitochondria. Insulin overdose increased hepatic p-FOXO1, CRM1, and 14-3-3 expression but decreased PEPCK and G6PC. FOXO1 shifted from nuclear to cytoplasmic localization in overdose groups, with partial nuclear recovery post-recovery. CONCLUSION: Insulin overdose leads to hypoglycemia, which in turn causes hepatocyte damage. Insulin overdose promotes the nuclear translocation of FOXO1 protein in hepatocytes, and this process shows high reproducibility in human cases of fatal insulin overdose. The coordinated changes in these indicators have significant value in detecting and diagnosing fatal cases of insulin overdose.