Alpha-1 antichymotrypsin is involved in astrocyte injury in concert with arginine-vasopressin during the development of acute hepatic encephalopathy - Fig 1

The growth and viability of human astrocytes cultured with plasma from mini-pigs with HE and the identification of candidate causative factors for HE by a 2-DE-based proteomic analysis (A) Experimental design: Mini-pig plasma was collected before the administration of α-amanitin and LPS, when encephalopathy occurred after the addition of toxins, and after treatment with extracorporeal circulation. (B) Astrocytes were cultured in medium with the collected plasma for 24 h. (C) Representative 2-DE gel images of silver-stained plasma proteins. These were based on LC-MS/MS data obtained from the NCBInr database using the MASCOT searching program. The labels 1 to 7 indicate the seven proteins that were upregulated before BAL treatment in comparison to normal plasma and which were downregulated after BAL treatment in comparison to before BAL treatment. (D) Astrocytes were incubated with various concentrations of ACT (0.01–0.5 mg/ml) for 24 h. Cell nuclei were counterstained with Hoechst 33342 with wavelengths of 350 nm excitation and 460 nm emission. Bars indicate the mean ± standard error of the mean (n = 5). *p < 0.05, in comparison to control (normal plasma).