Serum sphingosine-1 phosphate level is increased in patients with hepatitis B and displays a positive association with liver fibrosis

Abstract

Objective: To investigate the difference in serum sphingosine-1 phosphate (S1P) concentration between the HBV hepatitis patients and healthy controls, and its relevant association with serum liver fibrosis indicators.

Methods: A total of 28 HBsAg (+) HBeAg (+) Anti-HBc (+) hepatitis B patients, 42 HBsAg (+) Anti-HBe (+) Anti-HBc (+) hepatitis B patients, and 21 healthy subjects with normal liver function were included. Liquid chromatography-tandem mass method was used to detect the level of serum S1P.

Results: SerumS1P concentration of Anti-HBe (+) hepatitis B patients was higher than that of the control group (P=0.017) and HBeAg (+) patients (P=0.007). At the same time, there was no significant difference in the serum S1P concentration between HBeAg (+) hepatitis B patients and the control group (P>0.05). Moreover, serum S1P concentration was positively correlated with liver fibrosis indices, Collage Type IV Protein (r=0.264; P=0.011), and Chitosanase 3-like protein 1 (r=0.295; P=0.004).

Conclusions: Serum S1P level is increased in patients with hepatitis B and displays a positive association with liver fibrosis.

Keywords: Hepatitis B; chitosanase 3-like protein 1; collagen type IV protein; hepatic fibrosis; sphingosine 1-phosphate.

Figure 1

Flow chart of the inclusion and exclusion criteria in this study. 70 patients with hepatitis B were selected according to the principle of the method, and 42 hepatitis B patients with HBsAg (+) HBeAg (+) Anti-HBc (+) and 28 patients with HBsAg (+) Anti-HBe (+) Anti-HBc (+) were selected. In addition, 21 subjects were included in the control group of this study.

Figure 2

Serum concentrations of liver function indices in the control group and hepatitis B patients. A. The concentrations of ALT and AST in the HBsAg (+) HBeAg (+) Anti-HBc (+) group and the HBsAg (+) Anti-HBe (+) Anti-HBc (+) group were higher than in the control group; B. The concentrations of TP and ALB in the HBsAg (+) HBeAg (+) Anti-HBc (+) group and the HBsAg (+) Anti-HBe (+) Anti-HBc (+) group were lower than in the control group, while the difference of GLO between the three groups was not significant; C. The difference of TBIL, DBIL and TBA between the three groups were not significant. Data are the median with interquartile range. *P<0.05, **P<0.01, ***P<0.001, HBsAg (+) HBeAg (+) Anti-HBc (+) group and HBsAg (+) Anti-HBe (+) Anti-HBc (+) group compared to the control group respectively. (AST = alanine aminotransferase; ALT = aspartate transaminase; TP = total protein; ALB = albumin; GLO = globulin; TBIL = total bilirubin; DBIL = direct bilirubin; TBA = Total bile acid).

Figure 3

Serum concentrations of liver fibrosis indices in control group and hepatitis B patients. A. CHI3L1 of both hepatitis B groups were higher than the control group, and higher CHI3L1 levels in HBsAg (+) Anti-HBe (+) Anti-HBc (+) group than the HBsAg (+) HBeAg (+) Anti-HBc (+) group; B. The level of APRI in the HBsAg (+) HBeAg (+) Anti-HBc (+) group and the HBsAg (+) Anti-HBe (+) Anti-HBc (+) group were higher than in the control group, while the difference of FIB-4 between the three groups was not significant; C. Only the concentrations of C IV, HA, LN and PIIINP in the HBsAg (+) Anti-HBe (+) Anti-HBc (+) group were higher than in the control group, while the difference of C IV, HA, LN and PIIINP between the HBsAg (+) HBeAg (+) Anti-HBc (+) group and the control group were not significant Higher C IV and HA levels were noted in the HBsAg (+) Anti-HBe (+) Anti-HBc (+) group than in the HBsAg (+) HBeAg (+) Anti-HBc (+) group. Data are the median with interquartile range. *P<0.05, **P<0.01, ***P<0.001, HBsAg (+) HBeAg (+) Anti-HBc (+) group and the HBsAg (+) Anti-HBe (+) Anti-HBc (+) group compared to the control group respectively. #P<0.05, ###P<0.001, the HBsAg (+) HBeAg (+) Anti-HBc (+) group compared to the HBsAg (+) Anti-HBe (+) Anti-HBc (+) group. (CHI3L1 = chitinase-3-like protein-1; C IV = collagen type IV protein; HA = hyaluronic acid; LN = laminin; PIIINP = procollagen type III peptide; FIB-4 = fibrosis-4 score; APRI = aspartate aminotransferase platelet ratio index).

Figure 4

Serum concentrations of S1P in the control group and hepatitis B patients. S1P of HBsAg (+) Anti-HBe (+) Anti-HBc (+) group was higher than the control group and the HBsAg (+) HBeAg (+) Anti-HBc (+) group, while the difference in S1P between the HBsAg (+) HBeAg (+) Anti-HBc (+) group and control group was not significant. Data are the median with interquartile range. *P<0.05, HBsAg (+) Anti-HBe (+) Anti-HBc (+) group compared to the control group. ##P<0.01, HBsAg (+) Anti-HBe (+) Anti-HBc (+) group compared to the HBsAg (+) HBeAg (+) Anti-HBc (+) group. (S1P = sphingosine-1 phosphate).

Figure 5

Correlation between serum levels of S1P and the liver fibrosis indices. (A) APRI, (B) FIB-4, (D) HA, (E) LN and (F) PIIINP were uncorrelated with S1P, while (C) C IV (r=0.295, P=0.011) and (G) CHI3L1 (r=0.264, P=0.004) were positively correlated with S1P. (CHI3L1 = chitinase-3-like protein-1; C IV = collagen type IV protein; HA = hyaluronic acid; LN = laminin; PIIINP = procollagen type III peptide; FIB-4 = fibrosis-4 score; APRI = aspartate aminotransferase platelet ratio index; S1P = sphingosine-1 phosphate).

Figure 6

Correlation between serum levels of S1P and liver function indices. (A) ALT, (B) AST, (C) TP, (D) ALB, (E) GLO, (F) TIBL, (G) DBIL and (H) TBA were uncorrelated with S1P. (AST = alanine aminotransferase; ALT = aspartate transaminase; TP = total protein; ALB = albumin; GLO = globulin; TBIL = total bilirubin; DBIL = direct bilirubin; TBA = Total bile acid; S1P = sphingosine-1 phosphate).

Figure 7

Variation of liver fibrosis indices in L02 and LX-2 cells after S1P treatment. A. No difference in MMP9 and FN in L02 cells between control group and S1P; B. The levels of MMP9 and FN were increased in LX-2 cells after S1P treatment compared with control group. *P<0.05, **P<0.01, control group compared to the S1P group. (MMP9 = matrix metalloproteinase-9; FN = fibronectin; S1P = sphingosine-1 phosphate).