Bao-En Wang

HMGB1 Promotes the Synthesis of Pro-IL-1β and Pro-IL-18 by Activation of p38 MAPK and NF-κB Through Receptors for Advanced Glycation End-products in Macrophages

The high mobility group box-1 (HMGB1) protein and NALP3 inflammasome have been identified to play important roles in inflammation and cancer pathogenesis, but the relationships between the two and cancer remain unclear. The current study investigated the relationship between HMGB1 and the NALP3 inflammasome in THP-1 macrophages. HMGB1 was found unable to activate the NALP3 inflammasome and failed to induce the release of the IL-1β and IL-18 in THP-1 macrophages. HMGB1 was also found significantly enhanced the activity of ATP to induce IL-1β and IL-18 by the induction of increased expression of pro-IL-1β and pro-IL-18. This process was dependent on activation of RAGE, MAPK p38 and NF-κB signaling pathway. These results demonstrate that HMGB1 promotes the synthesis of pro-IL-1β and pro-IL-18 in THP-1 macrophages by the activation of p38 MAPK and NF-κB through RAGE. HMGB1 likely plays an important role in the first step of the release of the IL-1β and IL-18, preparing for other cytokines to induce excessive release of IL-1β and IL-18 which promote inflammation and cancer progression.

Expression of extracellular matrix genes in cultured hepatic oval cells: an origin of hepatic stellate cells through transforming growth factor beta?

Background: Hepatic oval cells, progenitor cells in the liver, can differentiate into hepatocytes and bile duct cells both in vitro and in vivo. Although hepatic stellate cells are another important cell component in the liver, less attention has been focused on the relationship between hepatic oval cells and hepatic stellate cells.

Antifibrotic Effects of a Recombinant Adeno-Associated Virus Carrying Small Interfering RNA Targeting TIMP-1 in Rat Liver Fibrosis

Elevated tissue inhibitor of metalloproteinase 1 (TIMP-1) expression contributes to excess production of extracellular matrix in liver fibrosis. Herein, we constructed a recombinant adeno-associated virus (rAAV) carrying siRNA of the TIMP-1 gene (rAAV/siRNA-TIMP-1) and investigated its effects on liver fibrosis in rats. Two models of rat liver fibrosis, the carbon tetrachloride and bile duct ligation models, were treated with rAAV/siRNA-TIMP-1. In the carbon tetrachloride model, rAAV/siRNA-TIMP-1 administration attenuated fibrosis severity, as determined by histologic analysis of hepatic collagen accumulation, hydroxyproline content, and concentrations of types I and III collagen in livers and sera. Levels of mRNA and active matrix metalloproteinase (MMP) 13 were elevated, whereas levels of mRNA and active MMP-2 were decreased. Moreover, a marked decrease was noted in the expression of α-smooth muscle actin, a biomarker of activated hepatic stellate cells (HSCs), and transforming growth factor-β1, critical for the development of liver fibrosis. Similarly, rAAV/siRNA-TIMP-1 treatment significantly alleviated bile duct ligation-induced liver fibrosis. Furthermore, this treatment dramatically suppressed TIMP-1 expression in HSCs from both model rats. These data indicate that the administration of rAAV/siRNA-TIMP-1 attenuated liver fibrosis by directly elevating the function of MMP-13 and diminishing activated HSCs. It also resulted in indirect decreased expression of type I collagen, MMP-2, and transforming growth factor-β1. In conclusion, rAAV/siRNA-TIMP-1 may be an effective antifibrotic gene therapy agent.

Primary isolated hepatic oval cells maintain progenitor cell phenotypes after two-year prolonged cultivation

BACKGROUND & AIMSnAlthough expandable hepatic progenitors provide renewable cell sources for treatment of hepatic disorders, long-term cultivation of hepatic progenitors may affect proliferation and differentiation abilities, and even initiate the formation of malignant cancer stem cells. This study aims to determine characteristics of primary cultured hepatic oval cells after prolonged cultivation in vitro.nnnMETHODSnHepatic oval cells isolated from rats fed with a choline-deficient, ethionine-supplemented diet were continuously propagated every 5-7 days, to 100 passages over two years. Hepatocytic differentiation was induced by sodium butyrate and characterized using western blot, periodic acid Schiff assays, albumin secretion and urea production. Proliferation capacity was evaluated using growth-curve and cell-cycle analysis; anchorage-independent growth and tumorigenicity were determined using soft agar and xenograft assay.nnnRESULTSnAfter 2 years of serial passages, hepatic oval cells with typical epithelial morphology continuously expressed OV-6, BD-1, BD-2, and Dlk as markers for hepatic progenitors, cytokeratin 19 as a cholangiocyte marker, and alpha-fetoprotein and albumin as hepatocyte markers. Furthermore, sodium butyrate could induce these cells to become glycogen-storage cells with the functions of albumin secretion and ureagenesis from ammonia clearance, indicating hepatocytic differentiation. Although proliferation slightly accelerated after the 50th passage, hepatic oval cells stayed diploid cells with features of chromosomal stability, which did not acquire anchorage-independent growth capacity and caused no tumor in immunodeficient mice, suggesting no spontaneous malignant transformation.nnnCONCLUSIONSnHepatic oval cells retain the progenitor cell features without spontaneous malignant transformation after prolonged cultivation, and thus may serve as an expandable cell source for future exploitation of stem cell technology.

EGF Suppresses the Initiation and Drives the Reversion of TGF‐β1‐induced Transition in Hepatic Oval Cells Showing the Plasticity of Progenitor Cells

Transforming growth factor‐β1 (TGF‐β1) induces hepatic progenitors to tumor initiating cells through epithelial‐mesenchymal transition (EMT), thus raising an important drawback for stem cell–based therapy. How to block and reverse TGF‐β1‐induced transition is crucial for progenitors’ clinical application and carcinogenic prevention. Rat adult hepatic progenitors, hepatic oval cells, experienced E‐cadherin to N‐cadherin switch and changed to α‐smooth muscle actin (α‐SMA) positive cells after TGF‐β1 incubation, indicating EMT. When TGF‐β1 plus EGF were co‐administrated to these cells, EGF dose‐dependently suppressed the cadherin switch and α‐SMA expression. Interestingly, if EGF was applied to TGF‐β1‐pretreated cells, the cells that have experienced EMT could return to their epithelial phenotype. Abruption of EGF receptor revealed that EGF exerted its blockage and reversal effects through phosphorylation of ERK1/2 and Akt. These findings suggest an important attribute of EGF on opposing and reversing TGF‐β1 effects, indicating the plasticity of hepatic progenitors. J. Cell. Physiol. 230: 2362–2370, 2015.

Increasing Newly Diagnosed Rate and Changing Risk Factors of HCV in Yanbian Prefecture, a High Endemic Area in China

Background The newly diagnosed rate of HCV infection is increasing in China. However, the risk factors have not been fully identified. Here, a survey was performed in Yanbian Prefecture, a high-endemic area in China. Methods We identified newly diagnosed HCV infection in 2007–2011, using the local National Disease Supervision Information Management System from the Chinese Center for Disease Control and Prevention. We determined the risk factors using a case-control survey by questionnaire. Results Yanbian Prefecture had a rapid increase in the yearly newly diagnosed rate of HCV infection from 32.6 to 72.1/100.000 from the year 2007 to 2011. People aged 50–64 years had a high HCV infection of 43.4%, but only 0.3% of cases were reported in those aged less than 20 years. Cosmetic treatment, family history, blood transfusion, and dental treatment were independent risk factors for HCV infection. Unexpectedly, cosmetic treatments [odd ratio (OR)u200a=u200a5.15, 95% confidence interval (CI)u200a=u200a2.31–11.48, Pu200a=u200a0.00] and family history (ORu200a=u200a4.68, 95% CIu200a=u200a2.67–8.75, Pu200a=u200a0.00) showed a higher risk than the conventional risk factors of blood transfusion (ORu200a=u200a4.49, 95% CIu200a=u200a1.95–10.37, Pu200a=u200a0.001) and dental treatment (ORu200a=u200a2.98, 95% CIu200a=u200a1.42–6.25, Pu200a=u200a0.00). To further analyze the intrafamilial transmission, we found that spouses of HCV patients had an increased risk for acquiring HCV (ORu200a=u200a5.75, 95% CI: 1.94–17.07), without significant association between either HCV RNA viral load (Pu200a=u200a0.29) or genotype (Pu200a=u200a0.43). Conclusions HCV infection was increased in Yanbian Prefecture. Cosmetic treatment was a higher risk factor than medical procedure. HCV infection had a clear family clustering phenomenon, especially between spouses.

Connective tissue growth factor induces hepatic progenitor cells to differentiate into hepatocytes

Connective tissue growth factor (CTGF) plays an important role in the proliferation of hepatic progenitors, however, little is known concerning the mechanism(s) through which it influences their differentiation. The differentiation of hepatic progenitors (WB-F344), either stimulated with recombinant CTGF or stably transfected with a CTGF overexpression plasmid, was investigated. Expression of the differentiation markers α-fetoprotein (AFP), albumin (ALB) and cytokeratin-19 (CK-19) was assessed. To confirm the effects of CTGF on progenitor differentiation, cells were treated with an inhibitor (WP631) of CTGF. Treatment of WB-F344 cells with recombinant CTGF for 24 h did not change the survival rate significantly, but the progenitors were enlarged with a decreased nuclear/cytoplasmic ratio. CTGF downregulated the expression of the fetal hepatocyte marker, AFP, while it upregulated the mature hepatocyte cell marker, ALB. The effect of CTGF overexpression plasmid on WB-F344 cell differentiation was consistent with a pattern of direct CTGF stimulation, including decreased AFP and increased ALB expression. Furthermore, the suppression of CTGF induction by an inhibitor was associated with significant inhibition of hepatic progenitor cell differentiation into hepatocytes. Importantly, we showed that differentiated WB-F344 cells by CTGF had in vitro functions characteristic of hepatocytes, including ALB production, glycogen storage and cytochrome P450 activity. Both recombinant CTGF and the CTGF overexpression plasmid induced hepatic progenitor differentiation into hepatocytes. This was suppressed by the CTGF inhibitor.

Different models of HBeAg seroconversion predicated by on-treatment ALT and HBV DNA profiles

Summary.u2002 Pretreatment alanine transaminase (ALT) elevation may be used as a predictor for higher hepatitis B e antigen (HBeAg) seroconversion in chronic hepatitis B patients. However, the role of dynamic changes of on‐treatment ALT for seroconversion is unknown. A total of 170 naïve HBeAg‐positive chronic hepatitis B patients were treated with a nucleoside/nucleotide analogues (NA), either lamivudine, adefovir, entecavir, or telbivudine, for at least 2u2003years and followed up for 1 more year. Clinical characteristics were detected and analysed at baseline and at 3‐month intervals. On‐treatment ALT predicted HBeAg seroconversion more accurately than baseline ALT. Among the patients with on‐treatment ALT ≤1u2003×u2003UNL, 1–≤2u2003× UNL, 2–≤5u2003×u2003UNL and >5u2003×u2003UNL, HBeAg seroconversion was 11.4, 5.4, 24.4 and 65.0% (odds ratiou2003=u20031.0, 0.4, 2.5 and 14.4, respectively), respectively. Moreover, two models/types of seroconversion were observed. Type I was characterized by rapidly decreased ALT and HBV DNA during the first 3‐month interval, but with high HBeAg reversion rate (50%) after consolidation treatment. Type II was a slow decreased DNA procedure accompanied by significant elevated ALT with less reversion (23%). Receiver operating characteristic curve analysis showed a 1.9‐fold increased ALT ratio (present visit ALT: previous visit ALT) accompanied by at least a 0.8 log decreased HBV DNA may be used to classify these two seroconversion types. We conclude that on‐treatment elevated ALT levels is a better predictor for seroconversion after NAs treatment, and HBV DNA profiles may help to identify different models of seroconversion.

Two patterns of alanine aminotransferase increase to predict long-term viral response in chronic hepatitis B patients: virus- or host-induced?

BACKGROUNDnSerum alanine aminotransferase (ALT) increase is a well-known phenomenon during interferon treatment for chronic hepatitis B. However, little is known about these increases during nucleoside/nucleotide treatment and the effects on long-term clinical outcomes.nnnMETHODSnA total of 170 treatment-naive hepatitis B e antigen (HBeAg)-positive chronic hepatitis B patients were treated with a nucleoside/nucleotide analogue for at least 2 years and followed up for 1 more year post-treatment. Clinical characteristics were detected and analysed at baseline and at every 3-month interval.nnnRESULTSnTwo patterns of ALT increase, virus- and host-induced, were detected. Virus-induced increases were characterized by a rapid increase in serum ALT and HBV DNA typically after 2 years of treatment, and were more common than host-induced ALT increases (15.9% versus 6.5%; P<0.05) with a median ALT increase of 5.7-fold the upper limit of normal (ULN). Host-induced ALT increases were characterized by moderately increased ALT (median 2.5-fold ULN) with a slow decrease in HBV DNA that occurred mainly in the first year of treatment (63.6%). Most importantly, host-induced increases were associated with favourable long-term treatment outcomes in HBV DNA undetectable rate (82% versus 0%), HBeAg seroconversion (82% versus 7%) and histological improvement. Moreover, interferon-γ-expressing T-helper cells were increased in patients with host-induced ALT increases.nnnCONCLUSIONSnTwo patterns of ALT increases may occur during nucleoside/nucleotide analogue treatment. Host induced ALT increases, accompanied by decreased HBV DNA, lead to better long-term clinical outcomes.