Abstract:

Objective To investigate the inhibitory effect of astragaloside Ⅳ（AS-Ⅳ） on cisplatin（CDDP）-induced liver injury in the mice，and to elucidate its possible mechanism. Methods Forty male C57BL/6 mice with body weights of 18-22 g were randomly divided into control group，model group，AS-Ⅳ group and adenosine 5'-monophosphate-activated protein kinase （AMPK） inhibitor （Compound C）+ AS-Ⅳ group. The mice in control group and model group were gavaged with the same volume of normal saline，and the drug was administered continuously for 9 d. The mice in AS-Ⅳ group and Compound C+AS-Ⅳ group were given AS-Ⅳ aqueous solution（150 mg·kg^-1·d^-1）， respectively. On the 6th day of experiment，the mice in Compound C+AS-Ⅳ group were intraperitoneally injected with Compound C （20 mg·kg^-1）， and on the 7th day，except for control group，the mice in other groups were intraperitoneally injected with 20 mg·kg^-1 CDDP to establish the mouse liver injury models，and the mice were sacrificed 48 h later. Serum and liver tissues were collected，and the levels of aspartate aminotransferase （AST） and alanine aminotransferase （ALT） in the serum of the mice，as well as the activities of superoxide dismutase（SOD） and catalase （CAT） and the levels of malondialdehyde（MDA） in the liver tissue of the mice in various groups were detected by kits. The pathomorphology of liver tissue of the mice in various groups were detected by HE staining. The expression levels of glutathione peroxidase 4 （GPX4）， ferritin heavy chain 1 （FTH1） and ferroptosis inhibitory protein 1 （FSP1） proteins in liver tissue of the mice in various groups were detected by immunohistochemical staining， and the expression levels of nuclear factor-E2-related factor 2 （Nrf2）， heme oxygenase-1 （HO-1） and AMPK proteins in liver tissue of the mice in various groups were detected by Western blotting method. Results Compared with control group， the levels of AST and ALT in serum of the mice in model group were increased （P<0.01）， the activities of SOD and CAT in the liver tissue were significantly decreased （P<0.01）， and the MDA level was increased （P<0.01）； compared with model group，the levels of AST and ALT in serum of the mice in AS-Ⅳ group were decreased （P<0.01）， the MDA level in the liver tissue was decreased （P<0.01）， and the activities of SOD and CAT were increased （P<0.01）； compared with AS-Ⅳ group （P<0.01）， the levels of AST and ALT in serum of the mice in Compound C+AS-Ⅳ group were increased （P<0.01）， the level of MDA in liver tissue was increased （P<0.05）， and the activities SOD and CAT were decreased（P<0.01）. The HE staining results showed that compared with control group， the liver damage degree of the mice in model group was enhanced，the hepatocyte arrangement was disordered， and some hepatocyte edema were increased； compared with model group， the liver morphology of the mice in AS-Ⅳ group returned to normal； compared with AS-Ⅳ group， the hepatocyte arrangement of the mice in Compound C+AS-Ⅳ group was disordered and the edges were blurred. The immunohistochemistry results showed that compared with control group， the expression levels of GPX4， FTH1 and FSP1 proteins in liver tissue of the mice in model group were decreased （P<0.05）； compared with model group， the expression levels of GPX4， FTH1 and FSP1 proteins in liver tissue of the mice in AS-Ⅳ group were increased （P<0.05）； compared with AS-Ⅳ group， the expression levels of GPX4， FTH1 and FSP1 proteins in liver tissue of the mice in Compound C+AS-Ⅳ group were decreased （P<0.05 or P<0.01）. The Western blotting results showed that compared with control group，the expression levels of Nrf2， HO-1 and AMPK proteins in liver tissue of the mice in model group were decreased （P<0.01）； compared with model group，the expression levels of Nrf2， HO-1 and AMPK proteins in liver tissue of the mice in AS-Ⅳ group were increased （P<0.01）； compared with AS-Ⅳ group，the expression levels of Nrf2， HO-1 and AMPK proteins in liver tissue of the mice in Compound C+AS-Ⅳ group were decreased （P<0.01）. Conclusion AS-Ⅳ can alleviate the CDDP-induced liver injury，and its mechanism may be related to the regulation of AMPK/Nrf2/HO-1 signal pathway and ferroptosis by AS-Ⅳ.

Key words: Astragaloside Ⅳ, Cisplatin, Liver injury, Ferroptosis, Adenosine 5'-monophosphate-activated protein kinase