Fraxin in Combination with Dexamethasone Attenuates LPS-Induced Liver and Heart Injury and Their Anticytokine Activity in Mice

Abstract

Background: Cytokine storm syndrome (CSS) is a major cause of morbidity and mortality in people suffering from hyperinflammatory status, which diverse etiological factors, including pathogens, therapeutic interventions, malignancies, and autoimmune disorders, can instigate. Since there is limited research on the antioxidant properties of fraxin and no studies have investigated its potential as an anticytokine storm agent, it is important to note that most studies have primarily focused on proinflammatory cytokines such as IL-1β, IL-6, and TNFα during cytokine storm. However, little research discusses the role of chemokines, particularly IL-8, during cytokine storms. Therefore, further investigation is warranted into the role of fraxin as an anticytokine storm agent and the involvement of IL-8 in cytokine storms. The present study examines the preventive efficacy of fraxin and the combination of fraxin and dexamethasone (FD) in mitigating lipopolysaccharide-induced systemic inflammation in mice caused by Escherichia coli, 055: B5.

Methods: Five groups of ten mice were randomly assigned: LPS only group (5 mg/kg, intraperitoneally i.p.), control (normal saline N.S. 1 ml/kg, i.p.), concentrations were selected based on previous literature, fraxin (120 mg/kg, i.p.), dexamethasone (5 mg/kg, i.p.), fraxin + dexamethasone (FD) (60 mg/kg + 2.5 mg/kg, i.p.), administered one hour before LPS injection (5 mg/kg,i.p.), animals were euthanized next day, and interleukin-8 (IL-8) was quantified in serum using an enzyme-linked immunosorbent assay. The liver and heart tissues underwent histopathological analysis to assess morphological changes. For data analysis using ANOVA and Tukey post hoc tests, the Kruskal-Wallis and Mann-Whitney U tests were employed to analyze the histological results.

Results: A significant decline in IL-8 levels was recorded in the treatment groups almost to the same degree (p < 0.001), and the percentage of inhibition of IL-8 for fraxin, dexamethasone, and FD was 93%.92.4%, and 93%, respectively, compared to the LPS-only group. Histopathological scores were significantly reduced in liver and heart tissue (P < 0.05).

Conclusions: All interventions used in this study significantly reduced interleukin-8 (IL-8) levels and reduced LPS-induced liver and cardiac damage. The combination (FD) did not result in an evident superiority of either agent. More research is required to identify the possible usefulness of these agents in treating hyperinflammatory diseases, such as cytokine storms, in future clinical practice.

Figure 1

The level of IL-8 in LPS-induced mice serum after 24 hours. Treatment with fraxin, dexamethasone, and fraxin + dexamethasone (FD); data are presented as mean ± SEM; ^∗highly significant P < 0.001 in comparison to LPS alone, and ^#highly significant P < 0.001 in comparison to control, n = 10. The red line indicates groups injected with LPS (5 mg/kg), 1 hour after treatment.

Figure 2

The histopathology of the liver (a). The control group liver section showing a normal structural appearance, central vein (red arrow), and sinusoids (yellow arrow), (b) lipopolysaccharide (LPS) only group liver section showing sinusoidal dilatation (yellow arrow), (c) (fraxin + LPS) near normal appearance of hepatocytes, (d) (DEX + LPS) liver section showing near-normal appearance, central vein, and threads of hepatocytes, and (e) (FD + LPS) liver section showing depletion of glycoprotein (blue arrow), hematoxylin and eosin, H&E stain, 40X.

Figure 3

Histopathology of heart section. (a) Control group showing normal cardiac muscle fiber, central nucleus with presence of henle space, (b) lipopolysaccharide (LPS) only group showing dispersed necrosis of cardiac muscle fiber (black arrow) with mild inflammatory cell infiltration (red arrow), dilatation of Henle space (pink arrow), (c) (fraxin + LPS) showing near-normal appearance, (d) (DEX + LPS) heart section showing near normal appearance with mild congestion (red arrow), and (e) (FD + LPS) heart section showing near normal appearance. Hematoxylin and eosin, H&E stain, 40X.

Figure 4

The histopathological scoring for all liver tissue treatment groups, with data presented as mean rank ± SEM. ^∗Treatment groups demonstrated statistical significance at p < 0.05 compared to the lipopolysaccharide LPS group and ^#significant at (p < 0.05) for LPS compared to the control, n = 5.

Figure 5

The histopathological scoring for all heart tissue treatment groups, with data presented as mean rank ± SEM. ^∗Treatment groups demonstrated statistical significance at p < 0.05 compared to lipopolysaccharide, LPS group, and ^#significant at (p < 0.05) for LPS compared to control, n = 5.