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