Astaxanthin regulates SLC7A11 to attenuate ferroptosis and autophagy levels in mice with non-alcoholic fatty liver disease

Ke Yue; Ji Wenjing; Liang Cancan; Wang Haikun; Cui Min; Yao Ping

doi:10.3969/j.issn.1671-6450.2025.06.018

Abstract：Objective To investigate the effect of astaxanthin(AST)intervention on the pathological process of mice with non-alcoholic fatty liver disease(NAFLD)and its molecular mechanism.Methods This study was conducted from June to November 2023 at the Animal Center of Xinjiang Medical University.A NAFLD mouse model was established using a high-fat diet,and mice were randomly divided into four groups:Control group,NAFLD group,NAFLD+Erastin group and NAFLD+AST group,Immunohistochemistry was used to detect the protein expression of SLC7A11 in liver tissues.Hematox-ylin-eosin(HE)staining was performed to observe inflammatory responses and structural changes in liver tissues.Enzyme-linked immunosorbent assay(ELISA)was conducted to measure the levels of blood lipids and inflammatory factors.Addition-ally,quantitative real-time polymerase chain reaction(RT-qPCR)and Western blot analyses were used to assess the expression levels of ferroptosis-related molecules(GPX4,HMOX1,SLC7A11)and autophagy-related molecules(LC3,p62).Results Immunohistochemistry showed that,compared with the blank control group,the expression of SLC7A11 protein in the NAFLD group decreased(P＜0.05).Compared with the NAFLD group,the expression of SLC7A11 protein in the NAFLD+Erastin group decreased(P＜0.05),while the expression of SLC7A11 protein in the NAFLD+AST group increased(P＜0.05).Compared with the NAFLD+AST group,the expression level of SLC7A11 protein in the NAFLD+Erastin group was lower(P＜0.05).The results of HE staining indicated that,compared with the blank control group,the inflammatory response in the liver tissue of the NAFLD group was exacerbated.When compared with the NAFLD group,the inflammatory response in the NAFLD+Erastin group was further aggravated,while that in the NAFLD+AST group was alleviated.AST can effectively re-duce the inflammatory response in the liver tissue.The inflammatory response in the NAFLD+AST group was significantly milder than that in the NAFLD+Erastin group.The detection of serum indicators showed that,compared with the blank control group,the levels of triglyceride(TG),total cholesterol(TC),and low-density lipoprotein Cholesterol(LDL-C)in the serum of mice in the NAFLD group were significantly increased(P＜0.05).Compared with the NAFLD group,the levels of TG,TC,and LDL-C in the NAFLD+Erastin group were all increased(P＜0.05),while the levels of TG,TC,and LDL-C in the NAFLD+AST group were all decreased(P＜0.05).The levels of TG,TC,and LDL-C in the NAFLD+Erastin group were significantly higher than those in the NAFLD+AST group(P＜0.05).Compared with the blank control group,the level of glutathione(GSH)in the NAFLD group was decreased(P＜0.05),while the levels of reactive oxygen species(ROS),malondialdehyde(MDA),tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6)were increased(P＜0.05).Compared with the NAFLD group,the level of GSH in the NAFLD+AST group was increased(P＜0.05),and the levels of ROS,MDA,TNF-α,and IL-6 were decreased(P＜0.05).In the NAFLD+Erastin group,except that the level of GSH was decreased(P＜0.05),the levels of ROS,MDA,TNF-α,and IL-6 were all increased(P＜0.05).Compared with the NAFLD+Erastin group,the level of GSH in the NAFLD+AST group was significantly increased(P＜0.05),and the levels of ROS,MDA,TNF-α,and IL-6 were decreased to some extent(P＜0.05).Molecular-level analysis revealed that qRT-PCR results showed that,compared with the blank control group,the ex-pressions of GPX4,HMOX1,p62,and SLC7A11 in the NAFLD group were downregulated(P＜0.05),while the expression of LC3B was upregulated(P＜0.05).Compared with the NAFLD group,the expressions of GPX4,HMOX1,SLC7A11,and p62 in the NAFLD+Erastin group were all decreased(P＜0.05),and the level of LC3B was increased(P＜0.05).In the NAFLD+AST group,the level of LC3B was decreased(P＜0.05),and the expressions of GPX4,HMOX1,SLC7A11,and p62 were all increased(P＜0.05).The expression levels of GPX4,HMOX1,SLC7A11,and p62 in the NAFLD+AST group were higher than those in the NAFLD+Erastin group(P＜0.05),while the expression of LC3B was decreased(P＜0.05).The results of Western Blot showed that compared with the blank control group,the expression of LC3B in the NAFLD group was upregulated(P＜0.05),and the protein expressions of GPX4,HMOX1,p62,and SLC7A11 were all downregulated(P＜0.05).Compared with the NAFLD group,the expression of LC3B in the NAFLD+Erastin group was upregulated(P＜0.05),and the expressions of GPX4,HMOX1,SLC7A11,and p62 were downregulated(P＜0.05).In the NAFLD+AST group,the expression of LC3B was downregulated(P＜0.05),and the expressions of GPX4,HMOX1,SLC7A11,and p62 were all upregulated(P＜0.05).The ex-pression of LC3B in the NAFLD+Erastin group was lower than that in the NAFLD+AST group(P＜0.05),and the expressions of GPX4,HMOX1,SLC7A11,and p62 were all increased(P＜0.05).Conclusion AST intervention may regulate the ferropto-sis and autophagy pathways by upregulating the expression of SLC7A11,thereby improving the pathological changes of liver tissues in NAFLD mice,reducing the levels of oxidative stress and inflammatory response,and providing potential new targets and innovative ideas for the treatment of NAFLD.