Protective Effects and Mechanisms of Babao Dan Capsules on ANIT-Induced Intrahepatic Cholestasis Liver Disease in Mice

Abstract Background Babao Dan(BBD), as a traditional Chinese medicine, is widely used in a variety of diseases, especially chronic liver diseases. The aim of this study is to observe the effects and possible mechanisms of Babao Dan(BBD) Capsules on α-Naphthyl isothiocyanate (ANIT) - induced intrahepatic cholestasis liver disease in mice. Methods Fifteen mice were randomly divided into three groups: control group, model group, and treatment group, with five in each group. Except for the mice in the control group, the mice in the other two groups were gavaged with a 1% α-Naphthyl isothiocyanate (ANIT) olive oil solution, and cholestasis models were established at a dose of 100 mg/Kg. Meanwhile, at the same time point, the control group was gavaged with the same amount of olive oil. After modeling, the control group, the model group and the treatment group were gavaged with the same amount of saline, saline and BBD saline suspension once daily for 10 days, respectively. On the 10th day, we took the blood preparation and liver samples, then we detected serum levels of liver function indexes(TBA, TBil, DBil, ALT, AST, ALP, GGT), liver fibrosis indexes(HA, LN, PC-Ⅲ, C-Ⅳ). Hepatic pathological examinations were observed by hematoxylin and eosin staining(HE), Masson staining and picric acid-Sirius red staining. The protein expression of TGF-β1 and Smad3 were detected by immunohistochemistry. The level of mRNA expression of TGF-β1 and Smad3 were detected by RT-PCR. Results Compared with the control group, serum levels of liver function indexes(TBA, TBil, DBil, ALT, AST, ALP, GGT) and liver fibrosis indexes(HA, LN, PC-Ⅲ, C-Ⅳ) in the model group were significantly increased (P < 0.01).Similarly, compared with control group, in the model group, HE staining showed that hepatocyte swelling, hepatocellular necrosis and destroyed hepatic lobular structure; Masson staining showed that a large amount of blue collagen fibers deposited in the liver tissue, forming fibrous septa and pseudolobule; Picric acid-Sirius red staining showed that type Ⅰ collagen and type Ⅲ collagen in liver tissue increased significantly, the total area of collagen increased significantly. The protein expression of TGF-β1 and Smad3 were significantly up-regulated; the mRNA expression of TGF-β1 and Smad3 was significantly up-regulated. Compared with the model group, the above indexes in treatment group were significantly decreased(P < 0.01). Conclusions Babao Dan(BBD) has significant therapeutical effects on intrahepatic cholestasis liver disease by inhibiting TGF-β/Smad signaling pathway.