Wei-Dong Pan

NP603, a novel and potent inhibitor of FGFR1 tyrosine kinase, inhibits hepatic stellate cell proliferation and ameliorates hepatic fibrosis in rats

Fibroblast growth factor 2 (FGF-2) and its main receptor FGFR1 have been shown to promote hepatic stellate cell (HSC) activation and proliferation. However, scant information is available on the anti-fibrogenic activity of FGFR1 inhibitors. The aim of this study was to assess the impact of a selective FGFR1 tyrosine kinase inhibitor NP603 on HSC proliferation and hepatic fibrosis. We demonstrated that rat primary HSCs secreted significant amounts of FGF-2, and its tyrosine phosphorylation of FGFR1 was attenuated by NP603. NP603 inhibited HSC activaton by measuring the expression of α-smooth muscle actin (α-SMA) and the production of type I collagen using ELISA. Furthermore, NP603 (25 μM) in vitro strongly suppressed HSC growth induced by FGF-2 (10 ng/ml) and FCS. This effect correlated with the suppression of extracellular-regulated kinase (ERK) activity and its downstream targets cyclin D1 and p21. In addition, PO NP603 (20 mg·kg(-1)·day(-1)) administration significantly decreased hepatic collagen deposition and α-SMA expression in CCl(4)-treated rats. Collectively, these studies suggest that selective blocking of the FGFR1-mediated pathway could be a promising therapeutic approach for the treatment of hepatic fibrosis.

Activation of Notch1 signaling by marrow-derived mesenchymal stem cells through cell-cell contact inhibits proliferation of hepatic stellate cells.

AIMS Bone marrow-derived mesenchymal stem cells (BMSCs) have been reported in many studies to reduce liver fibrosis. Apart from the paracrine mechanism by which the antifibrotic effects of BMSCs inhibit activated hepatic stellate cells (HSCs), the effects of direct interplay and juxtacrine signaling between the two cell types are poorly understood. The purpose of this study was to explore the underlying mechanisms by which BMSCs modulate the function of activated HSCs. MAIN METHODS We show here that BMSCs directly cocultured with HSCs significantly suppressed the proliferation and α-smooth muscle actin (α-SMA) expression of HSCs. Moreover, the Notch1 and Hes1 mRNA levels and the Hes1 protein level in cocultured HSCs were evidently higher than in other models. Blocking the Notch signaling pathway with Notch1 siRNA caused the increased expression of phospho-Akt and greater cell growth of cocultured HSCs. This effect was attenuated by the PI3K inhibitor LY294002. KEY FINDINGS In conclusion, our results demonstrated that BMSCs remarkably inhibited the proliferation of HSCs through a cell-cell contact mode that was partially mediated by Notch pathway activation. In addition, the PI3K/Akt pathway is involved in HSC growth inhibition by the Notch pathway. SIGNIFICANCE These findings demonstrated that BMSCs directly modulate HSCs in vitro via Notch signaling cascades. Our results may provide new insights into the treatment of hepatic fibrosis with BMSCs.

Activated hepatic stellate cells promote angiogenesis via interleukin-8 in hepatocellular carcinoma

BackgroundChemokines have been recognized as important modulators of angiogenesis, and they play critical roles in the development and metastasis of hepatocellular carcinoma (HCC), although their origins and latent molecular mechanisms remain elusive. The aim of this study was to investigate how activated hepatic stellate cells (a-HSCs) promote angiogenesis in HCC.MethodsA total of 22 HCC patients were enrolled randomly. We used immunohistochemistry, western blotting, and enzyme-linked immunosorbent assay (ELISA) to analyse the production of interleukin-8 (IL-8) in a-HSCs derived from HCC tissues. The angiogenic effects of IL-8 in vitro and in vivo were assessed by ELISA, real-time quantitative polymerase chain reaction, capillary tube formation assay, and chick embryo chorioallantoic membrane assay.ResultsThe present study showed that IL-8 was enriched predominantly in the tumour stroma of HCC tissues and was mainly derived from a-HSCs, rather than from hepatoma cells, in vivo and in vitro. Angiogenesis was most active at the invading edge, which was close to the a-HSCs. The angiogenic effect was dramatically attenuated by an IL-8 neutralizing antibody both in vitro and in vivo. Moreover, the IL-8 neutralizing antibody down-regulated Ser727-phosphorylated STAT3 levels in hepatoma cells treated with a-HSCs conditioned medium.ConclusionsThese findings reveal that a-HSCs within the stroma of HCC contribute to tumour angiogenesis via IL-8.

The expression of PEDF and VEGF in the gastric wall of prehepatic portal hypertensive rats.

BACKGROUND/AIMS Upper gastrointestinal bleeding of portal hypertension cases may result from gastric mucosal lesions due to portal hypertensive gastropathy. The pathological changes in the vessels of the gastric wall are very important in the pathogenesis of portal hypertensive gastropathy. However, the mechanisms of these pathological changes are not completely understood. In this study, we examined the expression levels of PEDF and VEGF in the gastric wall in rats with prehepatic portal hypertension. METHODOLOGY Eighteen healthy Wistar rats were randomly divided into groups A and B. Group A was used to establish the prehepatic portal hypertensive model and group B to evaluate a sham surgery. The VEGF and PEDF expression in the rat gastric wall were detected by immunohistochemical staining and western blotting. RESULTS VEGF and PEDF were mainly expressed in the basal layer of the mucosal glands. The expression levels of VEGF and PEDF in group A were higher than that in group B at 7, 10 and 14 days after surgery. The expression levels of VEFG and PEDF in group B did not show significant changes. CONCLUSIONS The results from the present study showed a significantly elevated expression of both VEGF and PEDF in the gastric walls during the development of portal hypertension. The expression of these proteins was mainly located in the basal layer of the gastric mucosa.

Beneficial effect of splenic artery ligation on bacterial translocation after major liver resection in rats

OBJECTIVE In major liver resection, bacterial translocation appears to be an important mechanism in the pathogenesis of spontaneous infection. This study was designed to investigate the effects of splenic artery ligation on bacterial translocation after major liver resection. MATERIALS AND METHODS Rats were divided into three groups: the sham operation group (SO group), the two-thirds partial hepatectomy group (PHx group) and the two-thirds partial hepatectomy plus splenic artery ligation group (PHx+Sp group). Bacterial translocation, endotoxemia, d-lactic acid and intestinal histology were analyzed among three groups. RESULTS The rate of bacterial translocation was higher in the PHx rats than in the SO rats (65.0% vs. 6.67%; P=0.001), so that in the PHx+Sp rats (25.0%; P=0.011). Endotoxemia was not evident in the SO rats (0pg/ml) and blood endotoxin levels decreased in the PHx+Sp rats (1.47pg/ml) compared with the PHx rats (4.05pg/ml, P<0.001). d-lactic acid was also higher in both the PHx and PHx+Sp rats compared with the SO rats (39.09mg/ml, 23.36mg/ml, and 1.68mg/ml; P<0.01). CONCLUSION Splenic artery ligation enhanced intestinal barrier function and diminished blood endotoxin levels and bacterial translocation in rats with major liver resection.

Experimental study of pancreaticojejunostomy completed using anastomotic chains

The most difficult, time-consuming, and complication-prone step in pancreaticoduodenectomy is the pancreaticojejunostomy step. The largest disadvantage of this kind of anastomosis is the high incidence of postoperative anastomotic leakage. Once pancreatic leakage occurs, the patient death rate can be very high. The aim of this study was to design a pancreaticojejunostomy procedure using anastomotic chains, which results in the cut end of the jejunum being attached to the pancreatic stump without suturing, and to evaluate the safety and efficacy of this procedure in domestic pigs. The pancreaticojejunal anastomotic chains had the following structures: the chains consisted of two braceletlike chains made of titanium, named chain A and chain B. The function of chain A was to attach the free jejunal end onto the pancreatic stump, whereas the function of chain B was to tighten the contact between the jejunal wall and the surface of the pancreatic stump to eliminate gaps between the two structures and ensure tightness that is sufficient to guarantee that there is no leakage of jejunal fluid or pancreatic juice. The following procedure was used to assess the safety and efficacy of the procedure: pancreaticojejunostomies were performed on ten domestic pigs using anastomotic chains. The time required to complete the pancreaticojejunal anastomoses, the pressure tolerance of the pancreaticojejunal anastomoses, the pig death rate, and the histopathological examinations of the pancreaticojejunostomy tissues were recorded. The average time required to complete the pancreaticojejunal anastomosis procedure was 13+/-2 min. The observed tolerance pressure of the pancreaticojejunal anastomoses was more than 90 mm H(2)O. All ten domestic pigs that underwent operations were still alive four weeks after the operations. Pathological examinations showed that the anastomotic surfaces were completely healed, and the pancreatic cutting surfaces were primarily epithelialized. In conclusion, the use of anastomotic chains in pancreaticojejunostomy procedures results in a decrease in or elimination of pancreatic leakage. In addition, the procedure is simple to perform, is not time-intensive, and appears to be safe in a pig model.