Objective

2

To explore the possible mechanism of

Shugan Jianpi Huoxue Formula

（疏肝健脾活血方， SJHF） in preventing and treating hepatic fibrosis.

Methods

2

Forty-eight C57BL/6 male mice were randomly divided into normal group， model group， SJHF group and CUG binding protein 1 （CUGBP1） knockout group， with 12 mice in each group. CUGBP1 knockout group was injected with CUGBP1 knockout adeno-associated virus （AAV9-shRNA-CUGBP1）， while the other three groups were injected with control adeno-associated virus （AAV9-control） through tail vein in volume of 100 μl with viral titers greater than 1×10

12

vg/ml. Four weeks later， except for the normal group， mice were injected with 2.5 ml/kg of 20% carbon tetrachloride （CCl

4

） intraperitoneally， once every three days for four weeks to induce hepatic fibrosis model； simultaneously， the SJHF group was given 33.3 g/（kg·d） of SJHF liquid by gavage， while the other three groups were given 100 ml/kg of distilled water by gavage， once a day for four weeks. Materials were collected 12 hours after the last medication，and the pathological morphology of the liver tissue was observed through HE staining， Masson staining and Sirius red staining. The levels of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in serum， and the hydroxyproline （HYP） content in the liver tissue were detected. The protein expression of type I collagen （CollagenⅠ）， α-smooth muscle actin （α-SMA）， phosphorylated Smad2 （p-Smad2）， phosphorylated Smad3 （p-Smad3）， Smad7， CUG binding protein 1 （CUGBP1）， as well as the mRNA expression of transforming growth factor β1 （TGF-β1）， Collagen Ⅰ， α-SMA， Smad7， and CUGBP1 in the liver tissue were detected.

Results

2

Compared to those in the normal group， the pathology of the liver tissue in the model group was characterized by disordered arrangement of hepatocytes， a large amount of infiltrated inflammatory cells and deposited collagen fibers， and obvious fibrous septa； the levels of ALT and AST in serum and the content of HYP in liver tissue increased； the CollagenⅠ， CUGBP1， p-Smad2， p-Smad3， and α-SMA protein expressions， as well as TGF-β1， CollagenⅠ， CUGBP1， and α-SMA mRNA expressions increased， while Smad7 protein and mRNA expressions decreased （

P

＜0.05 or

P

＜0.01）. Compared to those in the model group， he liver pathology in the SJHF group and the CUGBP1 knockout group showed a small amount of inflammatory cell infiltration， no obvious proliferation of fibrous tissue， and sparse fibrous septa； the levels of ALT and AST in serum， as well as the content of HYP in liver tissue decreased significantly， while all measured protein and mRNA expressions were improved （

P

＜0.05 or

P

＜0.01）. Compared to the SJHF group， the CUGBP1 knockout group had higher Smad7 mRNA expression， but no significant differences in other outcomes were found between the groups （

P

＞0.05）.

Conclusion

2

SJHF may inhibit the progression of hepatic fibrosis by inhibiting the expression of CUGBP1 in the liver tissue and blocking the subsequent transduction of TGF-β1/Smads signal.