Direct modulation of hepatocyte hepcidin signaling by iron.

BACKGROUND: Liver-secreted hepcidin is the systemic master switch of iron homeostasis and decreased levels of hepcidin are considered to cause iron overload not only in hereditary hemochromatosis but also in hemolytic anemia and chronic liver diseases. The regulation of hepcidin is complex and its response to iron is still not completely understood. AIM: To study the direct effect of iron on various established hepcidin signaling pathways in hepatoma cells or primary hepatocytes. METHODS: Hepcidin mRNA expression was studied by quantitative real-time (qRT)-PCR in the presence of various forms of iron including ferric ammonium citrate (FAC) in hepatoma cells (Huh7), murine primary hepatocytes and an established co-culture model of phorbol myristate acetate-differentiated THP-1 monocytes and Huh7 cells. To analyze hepcidin signaling, the response to bone morphogenetic protein 6 (BMP6), interleukin (IL)-6, IL-1ß, hypoxia and lipopolysaccharide (LPS) were studied. Hepcidin and small mothers against decapentaplegic 6 (SMAD6) mRNA levels were assessed by qRT-PCR and the expression of phosphorylated signal transducer and activator of transcription 3 (phospho-STAT3), STAT3, phospho-SMAD1/5/8 and SMAD1 proteins were analyzed by western blot. RESULTS: All iron III forms including FAC efficiently blocked hepcidin mRNA expression at non-toxic dosages in Huh7 cells or primary hepatocytes in a time and dose-dependent manner (P < 0.001; P < 0.05). Hepcidin blockage could be efficiently blunted by iron chelators salicylaldehyde isonicotinoyl hydrazone (SIH) and Desferal (P < 0.001). FAC also inhibited BMP6, hypoxia, IL-1ß and IL-6-mediated hepcidin induction (P < 0.001; P < 0.001; P < 0.05; P < 0.001), and FAC also inhibited LPS-mediated hepatic hepcidin induction in co-culture model (P < 0.001). Moreover, FAC reduced SMAD6 mRNA and p-SMAD1/5/8 protein expression at basal or upon stimulation by BMP6 (P < 0.05; P < 0.01), and FAC also reduced SMAD6 and p-SMAD1/5/8 expression under hypoxia (P < 0.01; P < 0.05). However, FAC has no significant effect on p-STAT3 protein expression at basal or upon stimulation by various stimuli. Notably, in the presence of the BMP/SMAD signaling pathway inhibitor LDN193189 Hydrochloride (LDN), FAC was unable to further decrease hepcidin, SMAD6 and p-SMAD1/5/8 expression compared with LDN alone. CONCLUSION: Iron directly blocks hepatocellular hepcidin signaling through the BMP/SMAD pathway but independent of STAT3. This mechanism may contribute to continued iron overload in many pathophysiological conditions ultimately causing a vicious cycle of continued hepcidin suppression.

Prevalence and Clinical Significance of Portal Vein Thrombosis in Patients With Cirrhosis and Acute Decompensation.

BACKGROUND & AIMS: Portal vein thrombosis (PVT) is a common and serious complication in patients with cirrhosis. However, little is known about PVT in patients with cirrhosis and acute decompensation (AD). We investigated the prevalence and clinical significance of PVT in nonmalignant patients with cirrhosis and AD. METHODS: We performed a retrospective study of 2 cohorts of patients with acute exacerbation of chronic liver disease who participated in the Chinese AcuTe on CHronic LIver FailurE study, established by the Chinese Chronic Liver Failure Consortium, from January 2015 through December 2016 (n = 2600 patients) and July 2018 through January 2019 (n = 1370 patients). We analyzed data on the prevalence, clinical manifestations, and risk factors of PVT from 2826 patients with cirrhosis, with and without AD. RESULTS: The prevalence of PVT in patients with cirrhosis and AD was 9.36%, which was significantly higher than in patients with cirrhosis without AD (5.24%) (P = .04). Among patients with cirrhosis and AD, 63.37% developed PVT recently (the first detected PVT with no indication of chronic PVT). Compared with patients without PVT, a significantly higher proportion of patients with PVT had variceal bleeding (47.33% vs 19.63%; P < .001) and patients with PVT had a significantly higher median serum level of D-dimer (2.07 vs 1.25; P < .001). Splenectomy and endoscopic sclerotherapy were independent risk factors for PVT in patients with cirrhosis and AD. The 1-year mortality rate did not differ significantly between patients with vs without PVT. CONCLUSIONS: In an analysis of data from 2826 patients with cirrhosis, a significantly higher proportion of those with AD had PVT than those without AD. PVT was associated with increased variceal bleeding, which would increase the risk for AD. Strategies are needed to prevent PVT in patients with cirrhosis, through regular screening, to reduce portal hypertension. ClinicalTrials.gov no: NCT02457637 and NCT03641872.

Acute-on-chronic liver failure exists in patients with hepatitis B virus-related decompensated cirrhosis.

OBJECTIVE: This research sought to verify whether acute-on-chronic liver failure (ACLF) develops in hepatitis B virus (HBV)-related cirrhotic patients with previous decompensation and to identify the similarity between assumed ACLF patients and those with ACLF that developed from compensated cirrhosis. METHODS: Patients with HBV-related cirrhosis were retrospectively screened and divided into the cirrhotic patients with first acute decompensation (AD) group and those with previous decompensation. Patients' characteristics, changes in laboratory results during hospitalization such as serum levels of total bilirubin (TB), creatinine (Cr) and white blood cell (WBC) counts, the Model for End-Stage Liver Disease (MELD) score and the 28-day and 1-year mortality rates were compared. RESULTS: Altogether 890 patients were enrolled and divided into the compensated cirrhotic group with first AD (n = 400; 157 of whom diagnosed as ACLF within 28 days after admission according to the European Association for the Study of the Liver-Chronic Liver Failure [EASL-CLIF] criteria) and those with previous decompensation (n = 490; of whom 143 met the ACLF criteria [assumed ACLF]). There was no significant difference in 28-day mortality between the assumed ACLF group with previous decompensation and ACLF group with first AD. The WBC count, TB and Cr levels, international normalized ratio and MELD score exhibited similar variations in both groups at days 1, 7 and 28; however, these values in both ACLF groups significantly differed from the non-ACLF group. CONCLUSION: HBV-related cirrhotic patients with previous decompensation who met the ACLF criteria had similar characteristic to ACLF patients with first AD.

Risk of different precipitating events for progressing to acute-on-chronic liver failure in HBV-related cirrhotic patients.

OBJECTIVE: Acute-on-chronic liver failure (ACLF) is a distinct syndrome that develops in patients with cirrhosis and acute decompensation (AD). This study focused on the precipitating events (PEs) of hepatitis B virus (HBV)-related cirrhotic patients diagnosed as ACLF based on the Chronic Liver Failure Consortium organ failure (CLIF-C OF) score. METHODS: Hospitalized patients with HBV-related cirrhosis and AD were retrospectively included. The patients' characteristics, laboratory test results, PEs, CLIF-C OF score and short-term prognosis were evaluated. RESULTS: Of the 890 patients enrolled 300 (33.7%) were diagnosed as ACLF and 590 (66.3%) without ACLF. ACLF patients had a higher incidence of PEs than those without ACLF. The ACLF patients were more prone to having PEs of bacterial infection (P < 0.001), HBV reactivation (P < 0.001), active alcoholism (P = 0.036) and superimposed hepatitis virus infection (P = 0.031), whereas portal vein thrombosis (P = 0.002) were less common in the non-ACLF group. ACLF patients with the top four single PEs had diverse types of organ failures. However, they shared a similar short-term prognosis. While in patients without PEs the ACLF group had higher systemic inflammation and deterirated outcomes compared with the non-ACLF group. CONCLUSIONS: PEs of bacterial infection, HBV reactivation, active alcoholism and superimposed hepatitis virus infection, but not GI hemorrhage or portal vein thrombosis, were risk factors for ACLF. There may be two types of patients with ACLF based on the differences in the clinical manifestation of the disease.

Pharmacokinetics of silybin nanoparticles in mice bearing SKOV-3 human ovarian carcinoma xenocraft.

The particle fabrication technique was used to fabricate monodisperse size and shape specific poly (lactide-co-glycolide) particles loaded with the silybin. Response surface methodology (RSM) using the central composite rotatable design (CCRD) model was used to optimize formulations of silybin nanoparticles. Further the optimized nanoparticles are characterized for particle size, zeta potential, surface morphology, entrapment efficiency, in-vitro drug release, silybin availability for tumor, plasma, lung, spleen, liver were determined. The significant findings were the optimal formulation of PLGA concentration 10 mg, PVA concentration 2000 and PET width of 6 gave rise to the EE of 88%, mean diameter of 223 nm and zeta potential of 25-mV. Release studies were investigated at pH 1.2 and pH 6.8. It was studied that lower the pH, faster the release of sylibin. The nanoparticles had~15-fold higher plasma exposure as measured by AUC contrasted to pure silybin. The nanoparticles had a 60% increase altogether tumor silybin presentation contrasted with pure silybin. Nanoparticles had higher silybin presentation in the spleen and liver contrasted with pure silybin suspension as expected for a nanoparticle formulation. The lung silybin presentation for the nanoparticle was additionally 2-fold higher than that of the pure silybin suspension. The results of pharmacokinetic parameters and oral bioavailability data exhibited that drug-nanoparticle complex could enhance the oral absorption of silybin and as well as the use of particles with smaller feature size may be preferred to decrease clearance by organs of the mononuclear phagocyte system.