Mechanisms of Gexia Zhuyu Decoction on anti-angiogenesis of hepatic fibrosis based on regulation of VEGF mRNA expression mediated by HIF-1α

CHEN Lan-yu1 MA Ji-zheng1 LIU Yong-mei1 ZHANG Yun

(1.Guang’anmen Hospital, China Academy of Chinese Medical Sciences, Beijing, China 100053)

【Abstract】 Objective To explore the mechanism of improving angiogenesis of hepatic fibrosis by Gexia Zhuyu Decoction (GZD) through the regulation of the mRNA expression of VEGF mediated by HIF-1α. Methods A total of 108 Wistar rats were randomly divided into normal group (n = 18), model group (n = 18), N-acetylcysteine (NAC) group (n = 18), high-dose GZD group (GD, n = 18), middle-dose GZD group (GZ, n = 18), and low-dose GZD group (GX, n = 18). Hepatic fibrosis model was established by intraperitoneal injection of 50% CCl4–olive oil solution (1 mL/kg) twice a week for nine weeks. Each group was administered while model established, until the rats were sacrificed. Normal group and model group were ig given sterile water 10 mL/(kg·d), NAC group was ig given NAC 0.1 g/(kg·d), GD, GZ, GX groups were given 26, 7.8, and 3.9 g/(kg·d) GZD by oral gavage. At 3, 6, and 9 weeks, rats in the corresponding groups were randomly sacrificed. Masson staining was used to make pathological specimens, immunohistochemical analysis of Col-Ⅳ and laminin was also performed, and real-time PCR was used to detect the mRNA expression of HIF-1α and VEGF. Western blotting was used to detect the protein expression levels of VEGF and VEGFR2. Results Compared with model group, NAC group and GD group significantly inhibited the expression of LN in the extracellular matrix at 9 weeks (P < 0.05). Both NAC and GD groups significantly inhibited the expression of extracellular matrix Col-Ⅳ, especially at 6 weeks and 9 weeks. NAC group, GD group, and GZ group can significantly inhibit the high expression of HIF-1α in liver tissue of rats with liver fibrosis (P < 0.05). At 6 weeks and 9 weeks of administration, NAC and GD groups significantly inhibited the high expression of VEGF mRNA in liver tissue (P < 0.05). Both GZD and NAC could inhibit the protein expression of VEGF and VEGFR2 in liver tissue. Conclusion GZD can regulate the expression of VEGF mRNA mediated by HIF-1α, which may be one of the key mechanisms of its anti-angiogenesis for hepatic fibrosis.

【Keywords】 Gexia Zhuyu Decoction; hepatic fibrosis; hypoxia inducible factor-1α; vascular endothelial growth factor (VEGF) ; collage type Ⅳ; laminin;


【Funds】 National Natural Science Foundation of China for Youths (81202633)

Download this article


    [1] Andrade Z A, Santana T S. Angiogenesis and schistosomiasis [J]. Mem Inst Oswaldo Cruz, 2010, 105 (4): 436–439.

    [2] Mejias M, Garci-Pras E, Tiani C, et al. Beneficial effects of sorafenib on splanchnic, intrahepatic, and portocollateral circulations in portal hypertensive and cirrhotic rats [J]. Hepatology, 2009, 49 (4): 1 245–1 256.

    [3] Llovet J M, Bruix J. Testing molecular therapies in hepatocellular carcinoma: The need for randomized phase Ⅱ trials [J]. J Clin Oncol, 2009, 27 (6): 833–835.

    [4] Siegel A B, Cohen E I, Ocean A, et al. Phase Ⅱ trial evaluating the clinical and biologic effects of bevacizumab in unresectable hepatocellular carcinoma [J]. J Clin Oncol, 2008, 26 (18): 2 992–2 998.

    [5] Wu Q, Liu Z C, Qi X D, et al. 膈下逐瘀汤对大鼠纤维化肝组织CTGF mRNA和PDGF-A蛋白表达的影响 [J]. China Foreign Medical Treatment, 2010 (24): 29–32 (in Chinese).

    [6] Zhang Y B, Jia Y, Niu Y C. Influence of Gexia Zhuyu soup on Smad4 mRNA expression of big mouse fiber liver organization [J]. Journal of Qiqihar University of Medicine, 2008, 29 (14): 1 669–1 670 (in Chinese).

    [7] Zhang Y B, Jia Y, Zhou L, et al. Effect of Gexia Zhuyu Tang on the expression of TIMP1 and MMP-2 in fibrotic liver in rats [J]. Chinese Journal of Basic Medicine in Traditional Chinese Medicine, 2008, 14 (9): 698–700 (in Chinese).

    [8] Zhang Y B, Jia Y, Niu Y C. 膈下逐瘀汤对大鼠纤维化肝组织a-SMA和MMP-2表达的影响 [J]. Journal of Qiqihar University of Medicine, 2008, 29 (15): 1 800–1 801 (in Chinese).

    [9] Yoshiji H, Kuriyama S, Yoshii J, et al. Vascular endothelial growth factor and receptor interaction is a prerequisite for murine hepatic fibrogenesis [J]. Gut, 2003, 52 (9): 1 347–1 354.

    [10] Bozova S, Elpek G O. Hypoxia-inducible factor-1 alpha expression in experimental cirrhosis: Correlation with vascular endothelial growth factor expression and angiogenesis [J]. APMIS, 2007, 115 (7): 795–801.

    [11] Ry H E, Poloni M, McNulty W, et al. Hypoxia inducible factor-1 alpha is positive factor in solid tumor growth [J]. Cancer Res, 2000, 60 (15): 4 010–4 015.

    [12] Jiang B H, Semenza G L, Bauer C, et al. Hypoxia inducible factor 1 levels vary exponentially over a physiologically relevant range of O2 tension [J]. Am JPhysiol, 1996, doi: 10.1152/ajpcell.1996.271.4.C1172.

    [13] Pfander D, Cramer T, Schipani E, et al. HIF-1α controls extracellular matrix synthesis by epiphyseal chondrocytes [J]. J Cell Sci, 2003, 116 (9): 1 819–1 826.

    [14] Pereira-Filho G, Ferreira C, Schwengber A, et al. Role of N-acetylcysteine on fibrosis and oxidative stress in cirrhotic rats [J]. Arquivos Gastroenterol, 2008, doi: 10.1590/S0004-28032008000200013.

    [15] Nakamura T, Akiyoshi H, Saito I, et al. Adenovirus mediated gene expression in the septal cells of cirrhotic rat livers [J]. J Hepatol, 1999, 30 (1): 101–106.

    [16] Wei W, Wu X M, Li Y J. 药理实验方法学 [M]. Beijing: People’s Medical Publishing House, 2010 (in Chinese).

    [17] Vanheule E, Geerts A M, Van Huysse J, et al. An intravital microscopic study of the hepatic microcirculation in cirrhotic mice models: Relationship between fibrosis and angiogenesis [J]. Int J Exp Pathol, 2008, 89 (6): 419–432.

    [18] Corpechot C, Barbu V, Wendum D, et al. Hypoxia induced VEGF and collagen I expressions are associated with angiogenesis and fibrogenesis in experimental cirrhosis [J]. Hepatology, 2002, doi: 10.1053/jhep.2002.32524.

    [19] Fine L G, Orphanides C, Norman J T. Progressive renal disease: The chronic hypoxia hypothesis [J]. Kidney Int Suppl, 1998, doi: 10.1046/j.1523-1755.1998.00853.x.

    [20] Prass K, Ruscher K, Karsch M, et al. Desferrioxamine induces delayed tolerance against cerebral ischemia in vivo and in vitro HIF-1: Mediator of physiological and pathophysiological responses to hypoxia [J]. J Cereb Blood Flow Metab, 2002, 22 (5): 520–525.

    [21] Coulon S, Heindryckx F, Geerts A, et al. Angiogenesis in chronic liver disease and its complications [J]. Liver Int, 2011, 31 (2): 146–162.

    [22] Corpechot C, Barbu V, Wendum D, et al. Hypoxia induced VEGF and collagen I expressions are associated with angiogenesis and fibrogenesis in experimental cirrhosis [J]. Hepatology, 2002, 35 (5): 1 010–1 021.

    [23] Moon J O, Welch T P, Gonzalez F J, et al. Reduced liver fibrosis in hypoxia-inducible factor-1 alpha-deficient mice [J]. Am J Physiol Gastrointest Liver Physiol, 2009, 296 (3): G582–G592.

    [24] Copple B L, Bai S, Burgoon L D, et al. Hypoxia-inducible factor-1α regulates the expression of genes in hypoxic hepatic stellate cells important for collagen deposition and angiogenesis [J]. Liver Int, 2011, 31 (2): 230–244.

    [25] Tang N, Wang L, Esko J, et al. Loss of HIF-1αin endothelial cells disrupts a hypoxia-driven VEGF autocrine loop necessary for tumorigenesis [J]. Cancer Cell, 2004, 6 (5): 485–495.

    [26] Copple B L, Kaska S, Wentling C. Hypoxia-inducible factor activation in myeloid cells contributes to the development of liver fibrosis in cholestatic mice [J]. JPharmacol Exp Ther, 2012, 341 (2): 307–316.

    [27] Zhang Q, Liu P, Zhang H W. Study on the patterns of TCM syndrome differentiation of 900 patients with post-hepatitic cirrhosis [J]. Chinese Journal of Integrated Traditional and Western Medicine, 2006, 26 (8): 694–697 (in Chinese).

This Article


CN: 12-1108/R

Vol 50, No. 02, Pages 449-456

January 2019


Article Outline


  • 1 Materials
  • 2 Methods
  • 3 Results
  • 4 Discussion
  • References