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

Jing Chen; Dian-Gang Liu; Guang Yang; Ling-Jian Kong; Ya-Ju Du; Hang-Yu Wang; Feng-Dong Li; Feng-Hua Pei; Ji-Tao Song; Yu-Jing Fan; Ai-Yun Liu; Xin-Hong Wang; Bao-Xin Li

doi:10.1177/1535370214532595

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

Exp Biol Med (Maywood). 2014 Aug;239(8):998-1006. doi: 10.1177/1535370214532595. Epub 2014 May 28.

Authors

Affiliations

¹ Department of Gastroenterology, the Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, China.
² Department of General Surgery, Xuan Wu Hospital, Capital Medical University, Beijing 100053, China Department of Pharmacology, Harbin Medical University, Harbin, Heilongjiang 150086, China.
³ Department of Gastroenterology, the First Hospital of Harbin, Harbin, Heilongjiang 150010, China.
⁴ Department of Pharmacology, Harbin Medical University, Harbin, Heilongjiang 150086, China libaoxinyaoli@126.com.

PMID: 24872431
DOI: 10.1177/1535370214532595

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 CCl₄-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 CCl₄ by intraperitoneal injection for six weeks. Six animal groups were studied: group 1: normal animals; group 2: CCl₄-induced liver fibrosis; group 3: CCl₄ + Endostar 20 mg/kg/d, six weeks; group 4: CCl₄ + Endostar 10 mg/kg/d, six weeks; group 5: CCl₄ + Endostar 20 mg/kg/d, four weeks; group 6: CCl₄ + 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 CCl₄-induced mice and collagen synthesis in HSCs in vitro. Therefore, this recombinant human endostatin is a promising compound for counteracting liver fibrosis.

Keywords: Carbon tetrachloride; Endostar; hepatic fibrosis; hepatic stellate cell; transforming growth factor β.