Background: Qizhu granule, a traditional Chinese medicine, has been widely used in clinic as a complementary and alternative medicine to treat liver fibrosis.However, the mechanism underlying its anti-hepatic fibrosis is still not clear. Liver fibrosis accompanied by liver sinusoidal pathological angiogenesis has been highlighted as novel therapeutic targets for the treatment of chronic liver disease. In this study, we investigated the mechanism of anti-capillarization of this herbal drug against liver fibrosis.
Materials and methods: The liver fibrosis rats model induced by 4-week of intervention with 40% CCl4 was employed in this study. Meanwhile, low, medium and high dose serum containing Qizhu granules were prepared. Liver tissues were acquired, and liver samples were subjected to histological studies.LSECs were isolated from liver fibrosis rats and were routinely cultured for 48h in low, medium and high dose of Qizhu granules-containing serum. The fenestration of LSECs in liver fibrosis rats were observed under scanning electron microscopy. The expression of the endothelial cell surface markers CD31, SE-1 and LSECs integrin ɑVβ3, FAK, p-FAK, Ras, MAPK, p-MAPK were measured by western blot.
Results: Compared with the control group, the loss of fenestration of LSECs in the model group increased. After intervention of Qizhu granule-containing serum, the fenestration of the structure of LSECs in liver fibrosis induced by CCl4 increased, especially in the high-dose Qizhu granules group. ompared with the control group, the expression levels of SE-1 and CD31 in LSECs in the model group were significantly increased (P < 0.05). Compared with the model group, the expression levels of SE-1 and CD31 in LSECs of rats with hepatic fibrosis induced by CCl4 decreased after the treatment of low, medium and high dose serum containing Qizhu granules (P < 0.05). Among them, the expression levels of SE-1 and CD31 in LSECs of Qizhu granules group decreased with the increase of dosage, showing a dose-dependent relationship to a certain extent. Compared with the normal control group, the expression of integrin ɑVβ3, Ras, p-FAK and p-MAPK protein increased in the LSECs of model group(P <0.05). After treatment with Qizhu granule-containing serum, the expression of integrin ɑVβ3, p-FAK and p-MAPK protein in LSECs of liver fibrosis rats induced by CCl4 were reduced (P <0.05), and the expression of FAK, Ras and MAPK protein decreased ( P> 0.05).
Conclusions: Qizhu granule could reduce the loss of fenestration of LSECs, transforming the cell phenotype of LSECs, and ameliorating the pathological remodeling of hepatic sinus capillarization in hepatic fibrosis induced by CCl4 in rats. It was found that Qizhu granules played an anti-fibrosis role by suppressing the expression of integrin ɑVβ3-FAK-Ras/MAPK signaling pathway of LSECs in CCl4-induced fibrosis rats.