黄芩苷通过调控HIF1A-GLS轴改善大鼠铜负荷肝豆状核变性的作用机制OA

Objective To investigate the mechanism of baicalin in improving copper-loaded hepatolenticular degeneration(HLD)rats based on the HIF1A-GLS axis.Methods(1)Differentially expressed genes in HLD were analyzed using GEO datasets.Targets of baicalin were screened via the SwissTargetPrediction database.Intersection of baicalin targets and HLD differentially expressed genes was identified as potential therapeutic targets.Protein-protein interaction(PPI)analysis was performed to screen key targets.GO functional and KEGG pathway enrichment analyses were conducted,and core targets were identified.Molecular docking validation was performed between baicalin and core targets.(2)HLD rat model was established by intragastric administration of CuSO4·5H2 O solution(300 mg·kg-1)once daily for 28 consecutive days.Fifty successfully modeled rats were randomly divided into model group,penicillamine group(90 mg·kg-1),and low-,medium-and high-dose baicalin groups(67.5,135,270 mg·kg-1),with 10 rats in each group.Another 10 SD rats served as normal group.Drugs were administered by gavage at a volume of 10 mL·kg-1 once daily for 28 consecutive days.Urinary copper,hepatic copper and serum copper levels were measured.Glutamic-pyruvic transaminase(GPT)and glutamic-oxaloacetic transaminase(GOT)levels in serum and liver tissue were detected.Liver index was calculated.Pathological changes of liver tissue were observed by HE staining.Protein expression levels of HIF1A and GLS in liver tissue were detected by immunohistochemistry.Protein expression of PDK1 and PDK3 was measured by Western Blot.mRNA expression levels of HIF1A,GLS,PDK1 and PDK3 were detected by real-time PCR.Results(1)A total of 1 284 HLD-related differentially expressed genes were identified,including 607 upregulated and 677 downregulated genes.Fifteen potential targets of baicalin against HLD were obtained,among which 8 were key targets.The potential targets were mainly involved in pathways such as complement and coagulation cascades,central carbon metabolism in cancer,and drug metabolism-cytochrome P450.HIF1A and GLS were identified as core targets,and baicalin showed stable binding with both.(2)Compared with the normal group,the model group exhibited significantly increased hepatic and serum copper levels(P＜0.05),while urinary copper showed no significant change(P>0.05).Liver index and GPT and GOT levels in serum and liver tissue were all elevated(P＜0.05).Disordered hepatic lobular structure,extensive hepatocyte necrosis,fibrotic lesions and inflammatory infiltration were observed.Protein and mRNA expression levels of HIF1A,GLS,PDK1 and PDK3 in liver tissue were all increased(P＜0.05).Compared with the model group,urinary copper levels were significantly elevated in the penicillamine group and medium-and high-dose baicalin groups(P＜0.05).Hepatic and serum copper levels were significantly reduced in all treatment groups(P＜0.05).Liver index and GPT and GOT levels were all decreased(P＜0.05).Hepatocyte necrosis and tissue fibrosis were alleviated to varying degrees.Protein and mRNA expression levels of HIF1A,GLS,PDK1 and PDK3 in liver tissue were all significantly reduced(P＜0.05).Conclusion Baicalin can reduce copper accumulation and improve liver function and pathological damage in HLD rats.Its mechanism may involve inhibiting the HIF1A-GLS axis to eliminate copper-overloaded hepatocytes and protect normal hepatocytes.