Human fibroblast growth factor-21 serves as a predictor and prognostic factor in patients with hepatitis B cirrhosis combined with adrenal insufficiency

Abstract

Hepatitis B cirrhosis is caused by liver cell necrosis, residual liver cell nodular regeneration, connective tissue hyperplasia and fiber formation, which frequently leads to adrenal insufficiency. Previous reports have demonstrated that human fibroblast growth factor (hFGF)-21 is a multifunctional protein that exhibits potential therapeutic value for metabolic diseases. The present study investigated the diagnostic value of hFGF-21 and analyzed the potential molecular mechanism in the progression of hepatitis B cirrhosis combined with adrenal insufficiency. Characteristics of cellular immunity and humoral immunity were analyzed in patients with hepatitis B cirrhosis combined with adrenal insufficiency (PhbA). Results demonstrated that expression levels of hFGF-21 were downregulated in plasma and liver cells isolated from clinical specimens. Plasma concentration levels of hFGF-21 were upregulated in prognostic PhbA. In vitro assays indicated that hFGF-21 treatment decreased the continuous deposition of extracellular matrix and reactive oxygen species in liver cells isolated from clinical specimens. Results also demonstrated that hFGF-21 treatment downregulated inflammatory cytokines. It was observed that hFGF-21 treatment downregulated nuclear factor (NF)-κB and Kruppel-like factor 6. Notably, transforming growth factor (TGF)-β, platelet-derived growth factor and epidermal growth factor levels were improved by hFGF-21 treatment. In conclusion, these results indicated that hFGF-21 inhibits inflammation by regulation of the NF-κB-mediated TGF-β signaling pathway, which may serve as a predictor and prognostic factor in PhbA.

Keywords: adrenal insufficiency; hepatitis B cirrhosis; human fibroblast growth factor-21; inflammation; nuclear factor-κB; transforming growth factor-β.

Figure 1.

Analysis of changes to hFGF-21 levels in PhbA. (A) Plasma concentration levels of hFGF-21 in PhbA and healthy volunteers. (B) Expression levels of hFGF-21 in liver cells isolated from clinical patients and healthy volunteers. **P<0.01. hFGF-21, human fibroblast growth factor-21; PhbA, patients with hepatitis B cirrhosis combined with adrenal insufficiency.

Figure 2.

Analysis of hFGF-21 expression levels in PPhbA and PhbA. (A) Plasma concentration levels of hFGF-21 between PPhbA and PhbA on day 30. (B) mRNA and protein expression levels of hFGF-21 in liver cells isolated from PPhbA and PhbA. **P<0.01. hFGF-21, human fibroblast growth factor-21; PPhbA, prognostic patients with hepatitis B cirrhosis combined with adrenal insufficiency; PhbA, patients with hepatitis B cirrhosis combined with adrenal insufficiency; PPhbA, patients who recovered form hepatitis B cirrhosis combined with adrenal insufficiency.

Figure 3.

Association of hFGF-21 plasma concentration with cellular immunity and humoral immunity in clinical patients. Relationship between concentration levels of hFGF-21 and (A) B lymphocyte level and (B) macrophagocyte level in patients during treatment. Association of hFGF-21 plasma concentration with percentage of (C) CD4⁺ and (D) CD8⁺ cells in serum in patients during treatment. hFGF-21, human fibroblast growth factor-21; CD, cluster of differentiation.

Figure 4.

Effects of hFGF-21 on inflammatory cytokine expression levels in liver cells isolated form clinical patients. Plasma concentration levels of (A) TNF-α, (B) IL-6, (C) IL-1β and (D) IL-8 in PhbA and healthy volunteers. (E) mRNA and (F) protein expression levels of inflammatory cytokines in cells treated with hFGF-21 isolated from patients and the PPhbA control group. **P<0.01. hFGF-21, human fibroblast growth factor-21; PhbA, patients with hepatitis B cirrhosis combined with adrenal insufficiency; PPhbA, patients who recovered form hepatitis B cirrhosis combined with adrenal insufficiency; TNF, tumor necrosis factor; IL, interleukin.

Figure 5.

Effects of hFGF-21 on inflammatory cytokine expression levels in renal epithelial cells isolated from clinical patients. (A) Gene expression levels of TNF-α, IL-6, IL-1β and IL-8 in renal epithelial cells isolated from clinical patients treated with hFGF-21 or not treated with hFGF-21. (B) Protein expression levels of TNF-α, IL-6, IL-1β and IL-8 in renal epithelial cells isolated from clinical patients treated with hFGF-21 or not treated with hFGF-21. **P<0.01. hFGF-21, human fibroblast growth factor-21; TNF, tumor necrosis factor; IL, interleukin.

Figure 6.

hFGF-21 regulates inflammatory cytokines through downregulation of the NF-κB-mediated TGF-β signaling pathway. (A) Effects of hFGF-21 on deposition of ECM and ROS expression levels in liver cells. (B) Effects of hFGF-21 on expression levels of TGF-β, NF-κB and KLF6 in liver cells. (C) Effects of hFGF-21 on expression levels of PDGF and EGF in liver cells. (D) Knockdown of NF-κB with Si-NF-κB increases TGF-β and KLF6 expression in liver cells. (E) Knockdown of NF-κB with Si-NF-κB suppresses PDGF and EGF expression in liver cells. (F) Effects of Si-NF-κB on protein expression levels of TNF-α, IL-6, IL-1β and IL-8 in liver cells. **P<0.01. hFGF-21, human fibroblast growth factor-21; NF, nuclear factor; TGF, transforming growth factor; ECM, extracellular matrix; ROS, reactive oxygen species; KLF6, Kruppel-like factor 6; PDGF, platelet-derived growth factor; EGF, epidermal growth factor; TNF, tumor necrosis factor; IL, interleukin; Si, small interfering RNA.