Impaired aminotransferase levels in long COVID-19 are associated with pro-inflammatory interleukin 8 (il-8) and vital neutrophil extracellular traps (netosis)

ABSTRACT

Background: The new coronavirus-19 (COVID-19) disease has been associated with a persistent symptomatic state after the acute phase, called Long COVID-19. This study aimed to find liver effects and their association with pro-inflammatory cytokines and percentage (%) of neutrophil extracellular traps (NETosis) in this stage Methods: Prospective, observational study including symptomatic subjects four months after COVID-19 diagnosis. Clinical, demographic, anthropometry data, and blood samples were collected at enrollment time. Non-contrast computed tomography of chest and abdomen was performed, and fatty liver was defined as liver attenuation <40 Hounsfield units. Alanine aminotransferase [ALT], aspartate aminotransferase [AST], glycemic and insulin blood levels were obtained. Interleukine (IL)-1β, IL-6, IL-8, IL-12, and Tumor necrosis factor (TNF-a) levels were measured in blood samples using flow cytometry. % of vital NETosis was measured in freshly isolated neutrophils using SYTOX and LIVE/DEAD dye correlated, ex vivo, with circulating cytokine levels. Continuous variables were analyzed by U de Mann-Whitney and nominal variables by Fisheŕs test. A logistic regression model analyzed the clinical association between liver attenuation. We evaluated the correlation between ALT, AST, cytokines, and NETosis using a linear model Results: 60 subjects were enrolled. The age average was 46.4 years old (standard deviation[SD]±13.1), and 32/60 (53.3%) were male. 25/60 (41.6%) reported severe COVID-19 during the acute phase. The prevalence of fatty liver was 17/60 (28.3%). 6 out of these 17 subjects had fatty liver before their COVID-19 diagnosis. Fatty liver was associated with high body mass index (p=0.02), waist circumference (p=0,01), insulin resistant (p=0,003), severe COVID-19 during acute phase (p=0,04), higher ALT (p<0.01), AST (p<0.01), IL-6 (p=0.02) and IL-12 (p=0.03). 12/60 (20%) patients had ALT elevation, although most of them, 8/12 (66.6%), did not have fatty liver. Both ALT and AST were correlated to IL-8 levels (R2=0.158, p<0.01), and IL-8 levels were significant correlated with % vital NETosis (R2=0.068, p=0.04) Conclusion: Fatty liver is prevalent and associated with severe COVID-19 during the acute phase and associated with high ALT, AST, IL-6, and IL-12 levels during the long COVID-19 phase. However, the ALT, AST and IL-8 elevations in the long COVID-19 might not be necessarily related to fatty liver. Therefore, the association between IL-8 and vital NETosis could be a possible pathway.