Mechanism of Zhizi Prescription in Protection of CCl4-induced Acute and Subacute Liver Injury in Mice / 中国实验方剂学杂志

ABSTRACT

ObjectiveTo investigate the protective effect of Zhizi prescription （ZZP） on carbon tetrachloride （CCl4）-induced acute and subacute liver injury and its mechanism. MethodAcute and subacute liver injury animal models were induced. C57 mice were randomly divided into a normal group， model group， obeccholic acid group， ZZP high-dose （0.5 g·kg-1） group， and ZZP low-dose （0.25 g·kg-1） group. According to the experiment design， the serum and liver tissue of mice were collected after the last administration. Hematoxylin-eosin （HE） and Sirius staining was used to observe the liver pathological changes. Serum alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total bilirubin （TBIL）， liver homogenate hydroxyproline （Hyp）， malondialdehyde （MDA）， and superoxide dismutase （SOD） levels were determined by kit. The levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in the liver tissue were determined by enzyme-linked immunosorbent assay （ELISA）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expressions of collagen 1A1 （Col1a1）， collagen 3A1 （Col3a1）， fibronectin （FN）， transforming growth factor β receptor Ⅱ （Tgfbr2） and α-smooth muscle actin （α-SMA） in the liver tissue. ResultIn terms of the acute liver injury， as compared with the normal group， the levels of ALT， AST， TBIL and MDA in the model group were significantly increased （P<0.01）， while the activity of liver SOD was significantly decreased （P<0.01）. Compared with model group， the ZZP high-dose and low-dose groups both significantly reduced the degree of liver cell injury， and protected the acute liver injury induced by CCl4. The ZZP high-dose group had a better effect than the ZZP low-dose group. In terms of the subacute liver injury， the levels of ALT， AST， MDA，TNF-α and IL-6 in the model group were significantly increased （P<0.01）， while the activity of liver SOD was significantly decreased （P<0.01）. As compared with the model group， liver Hyp content in the ZZP high-dose and low-dose groups was significantly decreased （P<0.01）， and the collagen deposition in liver of both groups was significantly reduced. The ZZP high-dose group also significantly down-regulated the mRNA expressions of α-SMA， Col1a1， Col3a1， FN， and Tgfbr2 in the liver of mice （P<0.05， P<0.01）. ConclusionZZP effectively protects the acute and subacute liver injury induced by CCl4， and the protective effect is proportional to its concentration. The mechanism may be related to the increase of the activity of antioxidant enzymes in the liver tissue， the decrease of the level of lipid peroxidation， and the inhibition of inflammatory response， thus reducing collagen deposition and improving early liver fibrosis.