Treatment with sodium (S)-2-hydroxyglutarate prevents liver injury in an ischemia-reperfusion model in female Wistar rats

Abstract

Background Ischemia-reperfusion (IR) injury is one of the leading causes of early graft dysfunction in liver transplantation. Techniques such as ischemic preconditioning protect the graft through the activation of the hypoxia-inducible factors (HIF), which are downregulated by the EGLN family of prolyl-4-hydroxylases, a potential biological target for the development of strategies based on pharmacological preconditioning. For that reason, this study aims to evaluate the effect of the EGLN inhibitor sodium (S)-2-hydroxyglutarate [(S)-2HG] on liver IR injury in Wistar rats. Methods Twenty-eight female Wistar rats were divided into the following groups: sham (SH, n = 7), non-toxicity (HGTox, n = 7, 25 mg/kg of (S)-2HG, twice per day for two days), IR (n = 7, total liver ischemia: 20 minutes, reperfusion: 60 minutes), and (S)-2HG+IR (HGIR, n = 7, 25 mg/kg of (S)-2HG, twice per day for two days, total liver ischemia as the IR group). Serum ALT, AST, LDH, ALP, glucose, and total bilirubin were assessed. The concentrations of IL-1β, IL-6, TNF, malondialdehyde, superoxide dismutase, and glutathione peroxidase were measured in liver tissue, as well as the expression of Hmox1, Vegfa, and Pdk1, determined by RT-qPCR. Sections of liver tissue were evaluated histologically, assessing the severity of necrosis, sinusoidal congestion, and cytoplasmatic vacuolization. Results The administration of (S)-2HG did not cause any alteration in the assessed biochemical markers compared to SH. Preconditioning with (S)-2HG significantly ameliorated IR injury in the HGIR group, decreasing the serum activities of ALT, AST, and LDH, and the tissue concentrations of IL-1β and IL-6 compared to the IR group. IR injury decreased serum glucose compared to SH. There were no differences in the other biomarkers assessed. The treatment with (S)-2HG tended to decrease the severity of hepatocyte necrosis and sinusoidal congestion compared to the IR group. The administration of (S)-2HG did not affect the expression of Hmox1 but decreased the expression of both Vegfa and Pdk1 compared to the SH group, suggesting that the HIF-1 pathway is not involved in its mechanism of hepatoprotection. In conclusion, (S)-2HG showed a hepatoprotective effect, decreasing the levels of liver injury and inflammation biomarkers, without evidence of the involvement of the HIF-1 pathway. No hepatotoxic effect was observed at the tested dose.

Acemetacin Induces Antioxidant Dysregulation, While Mefenamic Acid has a Hepatoprotective Effect Against Ischemia-Reperfusion Injury

“…This effect has been reported in studies assessing the toxicity of INDO, although the doses evaluated were higher 21,22 . Several reports from our research group have shown that these two biomarkers of biliary injury are not significantly affected by the induction of IR injury 13,23 . This could be explained by histological observations showing that the regions of cellular necrosis are focused on zone 3 of the hepatic acinus, close to the central vein.…”

“…To perform the calculation, a serum Alanine aminotransferase (ALT) activity of 1689±431 U LG 1 from a previous report of our group was considered, in which rats were subjected to the same protocol of IR injury induction 13 . By expecting a 40% decrease in the serum activity of ALT in the treated groups without a change in the standard deviation and considering a two-tailed "-value of 0.05 and a statistical power (1-$) of 80%, the value of n = 6 was obtained.…”

Ac2-26 attenuates hepatic ischemia-reperfusion injury in mice via regulating IL-22/IL-22R1/STAT3 signaling

“…Tissue sections were stained with hematoxylin and eosin in a standard manner dehydrated in graded alcohols and xylene and mounted with neutral gum. The morphological changes in hepatocytes were observed under light microscopy, and the Suzuki score ( Suzuki et al, 1993 ; Cienfuegos-Pecina et al, 2021 ) was used to evaluate the degree of liver injury: 0 = no congestion, no vacuolization, no necrosis; 1 = minimal congestion, minimal vacuolization, single-cell necrosis; 2 = mild congestion, mild vacuolization, necrosis <30%; 3 = moderate congestion, moderate vacuolization, necrosis <60%; 4 = severe congestion, severe vacuolization, necrosis >60%.…”