Exploring the Mechanism of Ruangan Granule on CCl₄-induced Hepatic Fibrosis in Rats Based on RhoA/ROCK Pathway

OBJECTIVE To explore the mechanism of Ruangan Granule against CCl₄-induced hepatic fibrosis in rats based on RhoA/ROCK pathway.

METHODS The rat hepatic fibrosis model was established by intragastric administration of 40% CCl₄ (2 mL·kg⁻¹)(CCl₄∶olive oil=2∶3), twice a week for 8 weeks. After the intervention of Ruangan Granule at 6 week for 8 weeks, Masson staining, organ index and biochemical indexes were used to evaluate the effect of Ruangan Granule on CCl₄-induced hepatic fibrosis in rats. RT-PCR, Western blotting and immunofluorescent tissue chemistry was used to detected the expression of Coll Ⅰ, α-SMA, RhoA, ROCK 1 mRNA and protein in rats.

RESULTS The organ index and biochemical analysis indicated that, compared with the model group, the liver index of rats was decreased in the middle dose group(P<0.01), and the spleen index was decreased in the high and middle dose groups(P<0.05 or P<0.01); high, middle and low dose of Ruangan Granule could increase the thymus index of rats(P<0.05 or P<0.01); Ruangan Granule could reduce the levels of AFP, ALT and AST in the serum(P<0.05 or P<0.01). The results of Masson staining showed that Ruangan Granule could significantly reduce the proliferation of collagen fibers in rat liver. RT-PCR, immune fluorescent tissue chemistry results showed that Ruangan Granule could significantly downregulate the expression of Coll Ⅰ, α-SMA, RhoA, and ROCK 1 mRNA and protein. Western blotting results showed the protein expression of Colli, α-SMA and Rock-1 were decreased in all groups. The protein expression of RhoA was decreased in middle and low dose groups of Ruangan Granule.

CONCLUSION The therapeutic effect of Ruangan Granule on CCl₄-induced hepatic fibrosis in rats may be related to the inhibition of RhoA/ROCK pathway.