Objective To investigate the effect of Shuganning
injection (SGN) in alleviating
drug -induced
cholestasis and the possible mechanisms involved.
Methods The
liver of
Sprague-Dawley rats was decellularized to prepare
collagen scaffolds, and then the scaffolds were recellularized with
human HepG2 cells to obtain the
tissue -engineered
liver (normal
control group ). The
tissue -engineered
liver was perfused with 10 μmol/L
chlorpromazine (CPZ) and
bile salt mixture to establish a model of
drug -induced
cholestasis (CPZ group), and the model was further treated with Shuganning
injection (10 3 -fold
dilution ) as the
injury protection group (SGN+CPZ group). The markers for hepatocellular
injury [
alanine aminotransferase (ALT),
aspartate aminotransferase (AST),
lactate dehydrogenase (LDH), and
alkaline phosphatase (ALP)] and the
antioxidant and
oxidative stress markers [
glutathione (GSH),
malondialdehyde (MDA),
superoxide dismutase (SOD), and
reactive oxygen species (ROS)] were measured for all groups, and the normal
control group , the CPZ group, and the SGN+CPZ group were compared in terms of the
mRNA and
protein expression levels of the
enzymes associated with
liver bile salt metabolism and the
enzymes associated with hepatic
cholestasis . HE
staining was performed to observe
liver pathology . A one-way
analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t -test was used for further comparison between two groups. Results Compared with the CPZ group, the SGN+CPZ group had significant reductions in the markers for hepatocellular
injury ALT, AST, LDH, and ALP (all P < 0.000 1), significant increases in the
oxidative stress markers GSH and SOD ( P < 0.000 1 and P < 0.001), and significant reductions in the markers MDA and ROS ( P < 0.000 1 and P < 0.001). Compared with the CPZ group, the SGN+CPZ group had significant reductions in the
mRNA expression levels of
cholesterol 7α-
hydroxylase (CYP7A1) and
sterol 12α-
hydroxylase (CPY8B1) in
hepatocytes (all P < 0.001) and significant increases in the
mRNA expression levels of farnesoid X receptor (FXR), small heterodimeric partner (SHP),
bile salt export pump (BSEP), and
multidrug resistance-associated protein 2 (MRP2) ( P < 0.000 1, P < 0.01, P < 0.000 1, and P < 0.000 1). HE
staining showed that compared with the CPZ group, the SGN+CPZ group had a significant reduction in
hepatocyte injury and a significant increase in the number of
cells . Conclusion Shuganning
injection can alleviate
drug -induced cholestatic
liver injury caused by
chlorpromazine , and it exerts a protective effect by activating FXR in
hepatocytes and increasing the expression of SHP to regulate
bile salt balance. It also inhibits CYP7A1 and
CYP8B1 to reduce the synthesis of hydrophobic
bile acids and upregulates the expression of BSEP and MRP2 to promote the excretion of
bile salts .