CYTOPROTECTIVE POTENTIAL OF CELL-FREE CRYOPRESERVED BIOLOGICAL AGENTS IN A MODEL OF NIMESULIDE-INDUCED LIVER INJURY

Abstract

Drug-induced liver injury remains a challenging issue in clinical pharmacology because of its variable manifestations and the lack of a specific diagnostic test. Nimesulide, which is used for short-term analgesia, may cause predominantly hepatocellular injury with a risk of acute liver failure. Therefore, the search for agents capable of simultaneously reducing oxidative stress, inflammation, and hepatocyte death remains highly relevant. Cell-free cryopreserved biological products contain peptides, cytokines, and growth factors and may provide cytoprotection. Of particular interest are placental and splenic extracts, as well as the secretome of mesenchymal cells. The aim of this study was to experimentally assess the hepatoprotective potential of cryopreserved placenta extract, cryopreserved spleen extract, and conditioned medium of mesenchymal stromal cells in rats with nimesulide-induced hepatopathy, based on cytolysis markers and the de Ritis ratio, in comparison with the reference drug Silibor. The study was conducted on 42 rats randomized into 6 groups of 7 animals each. Hepatopathy was induced by intragastric administration of nimesulide at a dose of 80 mg/kg body weight daily for 5 days. Cryopreserved placenta extract, cryopreserved spleen extract, conditioned medium of mesenchymal stromal cells, or Silibor were administered on days 1, 3, 5, 7, and 9. On day 10, aminotransferase activity and the de Ritis ratio were determined. It was found that nimesulide caused pronounced cytolysis: alanine aminotransferase activity increased to 2.39 ± 0.12 and aspartate aminotransferase activity to 2.80 ± 0.14 μmol/mL/h, compared with 1.06 ± 0.13 and 1.51 ± 0.11, respectively, in intact animals (p < 0.001). Silibor reduced alanine aminotransferase activity to 1.84 ± 0.14 (p = 0.012), whereas the decrease in aspartate aminotransferase activity was not statistically significant. Cryopreserved placenta extract reduced both enzymes to 1.37 ± 0.15 and 1.79 ± 0.27; cryopreserved spleen extract, to 1.57 ± 0.12 and 1.89 ± 0.12; and conditioned medium of mesenchymal stromal cells, to 1.39 ± 0.13 and 1.49 ± 0.10 (all p < 0.001–0.005 vs control). Compared with Silibor, the cell-free agents provided a more profound normalization, while conditioned medium showed the most complete restoration of aspartate aminotransferase activity, which is consistent with a broader effect on intracellular damage. On day 10, the de Ritis ratio generally did not differ significantly between the groups. Thus, the present study demonstrated that nimesulide reliably induces hepatocellular injury with increased aminotransferase activity. Silibor provides moderate cytoprotection, predominantly with respect to alanine aminotransferase. Cell-free cryopreserved agents reduce cytolysis markers more markedly, while conditioned medium of mesenchymal stromal cells demonstrates the most complete restoration of aspartate aminotransferase activity and may represent a promising therapeutic platform.