Endostar, a novel human recombinant endostatin, attenuates liver fibrosis in CCl₄-induced mice

Abstract

Decreasing hepatic fibrosis remains one of the major therapeutic challenges in hepatology. The present study aims to evaluate the effect of Endostar on both CCl-induced liver fibrosis in mice and a hepatic stellate cell (HSC) line. Two main models were studied: (i) a liver fibrosis model was induced in BALB/c mice using CCl by intraperitoneal injection for six weeks. Six animal groups were studied: group 1: normal animals; group 2: CCl-induced liver fibrosis; group 3: CCl + Endostar 20 mg/kg/d, six weeks; group 4: CCl + Endostar 10 mg/kg/d, six weeks; group 5: CCl + Endostar 20 mg/kg/d, four weeks; group 6: CCl + Endostar 10 mg/kg/d, four weeks corresponded to different Endostar doses and duration of administration. Liver fibrosis was evaluated by histopathological staining and liver hydroxyproline content. Expressions of collagen type I, α-smooth muscle actin (α-SMA), TGF-β1 and VEGFR were measured by real-time polymerase chain reaction (PCR). (ii) A liver cell model. HSC-T6 cells were cultured with or without Endostar for 12 h or 24 h. Expressions of collagen type I, α-SMA, and TGF-β1 were measured by real-time PCR. Collagen I and transforming growth factor β1 (TGF-β1) contents in cell supernatant were measured by enzyme-linked immunosorbent assay. As compared to the group without Endostar, liver fibrosis scores and hydroxyproline content were decreased in both Endostar groups (P < 0.05). Moreover, Endostar inhibited the hepatic expression of α-SMA, TGF-β1, Collagen-1, VEGFR1, and VEGFR2 mRNA (P < 0.05). In the HSC-T6 cell line model, Endostar profoundly inhibited the expression of α-SMA, Collagen-1, and TGF-β1 mRNA. Expressions of Collagen-1 and TGF-β1 protein were decreased in the Endostar group as compared to the normal controls in the supernatant of HSC-T6 cells (P < 0.05). Endostar decreased both liver fibrosis in CCl-induced mice and collagen synthesis in HSCs in vitro. Therefore, this recombinant human endostatin is a promising compound for counteracting liver fibrosis.

Hepatoprotective effect of oral application of a silymarin extract in carbon tetrachloride-induced hepatotoxicity in rats

“…TGF-β leads to an autocrine induction of fibrogenic gene expression in HSC. SE treatment reduced the CCl 4 -induced expression of TGF-β and collagen-I which play crucial roles in liver fibrosis also in the CCl 4 model [6,7,9,16]. In our study, a remarkable decrease of pro-fibrogenic and pro-inflammatory mRNAs was observed, leading to the conclusion that the used silymarin extract has protective effects on the formation of fibrosis after acute liver injury induced by models of liver injury [2,4,14,21,23].…”

“…Furthermore, anti-inflammatory and anti-fibrotic properties have been described (reviewed in [1]). While a few studies have investigated the effectiveness of silymarin in a chronic CCl 4 model upon oral application and with regards to the fibrogenic reaction [6,16]. We for the first time studied the acute effects of orally administered milk thistle extract on early events in fibrosis development.…”

Vatalanib, a tyrosine kinase inhibitor, decreases hepatic fibrosis and sinusoidal capillarization in CCl4-induced fibrotic mice

“…A2547; Sigma-Aldrich; Merck Millipore, Darmstadt, Germany) and CD34 (cat no. ab81289; Abcam, Cambridge, MA, USA) and an avidin-biotin complex immunoperoxidase method (15). Following antigen retrieval by immersion in EDTA (1 mmol/l, pH 8.0) and boiling for 15 min, sections were pre-blocked with normal goat serum for 10 min, and incubated for 2 h at room temperature with specific antibodies (α-SMA, dilution 1:200; CD34, dilution, 1:200).…”

“…Anti-angiogenesis treatment may be a therapeutic approach in liver fibrosis and portal hypertension (11). It has been previously reported that anti-angiogenesis drugs, including sorafenib, sunitinib, and Endostar, which are used in the treatment of carcinoma, inhibit hepatocellular carcinoma and liver fibrosis (12–15). …”

Human recombinant endostatin Endostar attenuates hepatic sinusoidal endothelial cell capillarization in CCl4-induced fibrosis in mice

“…In a previous study, it was demonstrated that Endostar decreased liver fibrosis and necrosis in a mouse model of liver fibrosis induced by CCl 4 and inhibited collagen synthesis in the HSC-T6 rat stellate cell line in vitro (9). The present study aimed to further explore the antifibrogenic effects of Endostar by investigating the impact of Endostar on SEC phenotype in CCl 4 -induced fibrotic mice, and to better understand the mechanisms underlying this action.…”