Do Semaphorins Play a Role in Development of Fibrosis in Patients with Nonalcoholic Fatty Liver Disease?

Lara Šamadan, Neven Papić, Maja Mijić, Ivana Knežević Štromar, Slavko Gašparov, Adriana Vince, Lara Šamadan, Neven Papić, Maja Mijić, Ivana Knežević Štromar, Slavko Gašparov, Adriana Vince

Abstract

Nonalcoholic fatty liver disease (NAFLD) is associated with systemic changes in immune response linked with chronic low-grade inflammation and disease progression. Semaphorins, a large family of biological response modifiers, were recently recognized as one of the key regulators of immune responses, possibly also associated with chronic liver diseases. The aim of this study was to identify semaphorins associated with NAFLD and their relationship with steatosis and fibrosis stages. In this prospective, case-control study, serum semaphorin concentrations (SEMA3A, -3C, -4A, -4D, -5A and -7A) were measured in 95 NAFLD patients and 35 healthy controls. Significantly higher concentrations of SEMA3A, -3C and -4D and lower concentrations of SEAMA5A and -7A were found in NAFLD. While there was no difference according to steatosis grades, SEMA3C and SEMA4D significantly increased and SEMA3A significantly decreased with fibrosis stages and had better accuracy in predicting fibrosis compared to the FIB-4 score. Immunohistochemistry confirmed higher expression of SEMA4D in hepatocytes, endothelial cells and lymphocytes in NAFLD livers. The SEMA5A rs1319222 TT genotype was more frequent in the NAFLD group and was associated with higher liver stiffness measurements. In conclusion, we provide the first evidence of the association of semaphorins with fibrosis in patients with NAFLD.

Keywords: NAFLD; SEMA3A; SEMA3C; SEMA5A; SEMA7A; cirrhosis; fibrosis; nonalcoholic fatty liver disease; semaphorins; steatosis.

Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

Figures

Figure 1
Figure 1
Serum concentrations of semaphorin SEMA3A (a), SEMA3C (b), SEMA4A (c), SEMA4D (d), SEMA5A (e) and SEMA7A (f) measured by ELISA in healthy controls and in patients with NAFLD, stratified by steatosis grade and fibrosis stage. Data are presented as median with interquartile range. The p-values are calculated by Mann–Whitney U-test or Kruskal–Wallis test, as appropriate.
Figure 2
Figure 2
Spearman correlation correlogram. The strength of the correlation between two variables is represented by the color at the intersection of those variables. Colors range from dark blue (strong negative correlation; r = −1.0) to red (strong positive correlation; r = 1.0). Results were not displayed if p > 0.05.
Figure 3
Figure 3
The ROC curve analysis of serum semaphorin concentrations for discrimination of NAFLD (a) and progressed fibrosis (b). Shown are AUC, area under the curve, with corresponding 95% confidence intervals.
Figure 4
Figure 4
Immunohistochemical staining of SEMA4D in healthy (a) and NAFLD livers (b–d). While there is no expression of SEMA4D marker in hepatocytes of healthy liver tissue ((a), magnification 60×), in NAFLD liver without significant fibrosis ((b), magnification 60×; in upper right corner hematoxylin-eosin stain showing fat accumulation), there is a moderate expression in hepatocytes, endothelial cells and lymphocytes. In cirrhotic liver ((c,d), magnification 60×), SEMA4D showed a stronger signal. In the upper right quadrant in panel (b), Masson’s trichrome staining of cirrhotic liver is shown.
Figure 5
Figure 5
Association of SEMA5A rs1319222 and rs433755 genotype with SEMA5A serum concentrations (a,c) and liver stiffness (kPa) and controlled-attenuation parameter (db/m) measurements (b,d). Data are presented as median with interquartile range. The p-values are calculated by Kruskal–Wallis test, and significant differences calculated by Dunn’s multiple comparisons test.

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