Dantao formula alleviates hepatic fibrosis and portal hypertension via modulation of the cAMP/PKA/ROCK signaling pathway in hepatic stellate cells.

ETHNOPHARMACOLOGICAL RELEVANCE: The Dantao Formula (DTF) consists of Salvia miltiorrhiza Bunge (Danshen) and Prunus persica (L.) Batsch seeds (Taoren), which were the components of promoting circulation and removing stasis in the anti-fibrotic herbal--- Fuzheng Huayu Formula (FZHY). It has demonstrated efficacy in alleviating hepatic fibrosis and portal hypertension (PH); however, its pharmacological mechanisms remain unclear.

AIM OF THE STUDY: The aim is to investigate the effects and molecular mechanisms of DTF on liver fibrosis and PH.

MATERIALS AND

METHODS

In vivo experiments were evaluated using a CCl4-induced mouse model with rivaroxaban (RIVA) as a drug control. PH was assessed by direct puncture method. Fibrosis was assessed by Sirius red staining and hydroxyproline. Network pharmacology analysis predicted the potential molecules or signal mediators. In vitro, an LX-2 cells was activated with ET-1, and Y-33075 used as a drug control. ELISA, Western blot, immunohistochemistry and immunofluorescence were conducted to assess the target expression of ET-1, ENDRA, ROCK, cAMP, PKA, MLC and p-MLC.

RESULTS

DTF or RIVA could alleviate liver fibrosis and PH compared to the model group. Network pharmacology analysis suggested that cAMP/PKA/ROCK signaling pathway acted as a key target in DTF. In vivo, DTF or RIVA suppressed ET-1, EDNRA and ROCK expression, enhanced cAMP and PKA expression. In vitro, DTF or Y-33075 attenuated the activation of LX-2 cells induced by ET-1, down-regulated ROCK and p-MLC expression, up-regulated cAMP and PKA expression.

CONCLUSIONS

DTF alleviates liver fibrosis and PH by regulating the cAMP/PKA/ROCK signaling pathway in activated HSCs.