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Previous investigations have established that ferulic acid demonstrates anti-fibrosis effects and could inhibit the activation of hepatic stellate cell. Injury, proliferation and capillarization of hepatic sinusoidal endothelial cell are important forms of angiogenesis during liver fibrosis, and is important for liver fibrosis regression. This study investigates the effects of ferulic acid on sinusoidal capillarization and explores its anti-fibrotic mechanism. In vitro, SK-HEP-1 cells were injured by cobalt chloride, while human hepatic sinusoidal endothelial cells (HHSECs) were induced by endothelial cell growth supplement (ECGs). Cell viability was detected using thiazolyl blue tetrazolium bromide (MTT); nitric oxide (NO) and nitric oxide synthase (NOS) were visualized through the fluorescence probe method; antiangiogenesis effects were assessed using the tube formation assay and transgenic zebrafish (VEGFR: GFP). Animal experiments complied with the regulations of the Animal Ethics Committee of Shanghai University of Traditional Chinese Medicine (approval No. PZSHUTCM2503280008). Mice were received intraperitoneal injection of carbon tetrachloride (CCl4) for 6 weeks to induce hepatic fibrosis and then treated with 10 mg·kg-1 of sorafenib, 50 and 100 mg·kg-1 of ferulic acid for 3 weeks from the fourth week. Liver inflammation and fibrosis were assessed through serum liver function [alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin (ALB)] tests, H & E and Sirius red staining, hydroxyproline (HYP) quantification, and the immumohistochemical staining of α-smooth muscle actin (α-SMA). Sinusoidal fenestrations were observed via scanning electron microscopy (SEM), and the phenotype of liver sinusoidal endothelial cells were labeled by platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31) and lymphatic vessel endothelial hyaluronan receptor 1 (LYVE1). Results showed that ferulic acid protected SK-HEP-1 cells from CoCl2-induced hypoxic injury, significantly increased NO and NOS levels. It suppressed the proliferation and tube formation of HHSEC, and inhibited angiogenesis of VEGFR: GFP zebrafish model. In vivo experiments, ferulic acid reduced serum AST/ALT and HYP levels and increased ALB. Histopathological analyses confirmed that it reduced inflammation and fibrosis, downregulated α-SMA expression, ameliorated sinusoid capillarization and downregulated the expression of CD31 and LYVE1. These findings indicate that ferulic acid can inhibit liver fibrosis through alleviating sinusoidal capillarization.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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