Inhibitory Effect of Gandou Decoction Combined with Penicillamine on Liver Cuproptosis in Wilson Disease Model
Objective To investigate the inhibitory effect of Gandou Decoction combined with penicillamine on cuproptosis in the liver of toxic milk(TX)mice,which serve as a model for Wilson's disease(WD).Methods Using dilute lethal(DL)mice as the normal control group,and TX mice were randomly divided into the model group,Gandou Decoction group,penicillamine group,and Gandou Decoction combined with penicillamine group,with 8 mice in each group.The normal control group and the model group were given 0.2 mL/10 g-1·d-1 of 0.9％NaCl solution,while the Gandou Decoction group,penicillamine group and Gandou Decoction combined with penicillamine group were separately given 0.2 mL·10 g-1·d-1 of Gandou Decoction,penicillamine 0.1 g·kg-1·d-1 and Gandou Decoction 0.2 mL·10 g-1·d-1 combined with penicillamine 0.1 g·kg-1·d-1 by gavage for 30 consecutive days.Biochemical methods,inductively coupled plasma mass spectrometry(ICP-MS),HE staining,transmission electron microscopy,real-time quantitative PCR,Western Blot and immunohistochemistry were used to determine the serological indexes and hepatic copper content,pathological changes,mitochondrial ultrastructure,cuproptosis-related genes and proteins,and the expression level of lipoic acid(LA)in each group of mice.Results Compared with the control group,the model group exhibited elevated serum ALT and AST levels,LDH and SOD activities,and hepatic tissue copper content(P＜0.05).Additionally,the model group showed localized fragmentary necrosis of the liver,an increased number of inflammatory cells(P＜0.01),significant mitochondrial damage,decreased expression levels of cuproptosis-related genes mRNA,iron-sulfur cluster proteins,thioctanoylated proteins,and LA(P＜0.05).Furthermore,the expression levels of dihydrolipoic acid transacetylase(DLAT)dimer and heat shock protein 70(HSP70)increased(P＜0.01).Compared with the model group,the treatment groups exhibited a decrease in serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels,lactate dehydrogenase(LDH)and superoxide dismutase(SOD)activities,as well as hepatic tissue copper content(P＜0.05).Additionally,there was a reduction in hepatic pathological injury and a decrease in the number of inflammatory cells(P＜0.01).Moreover,the mitochondrial structure was restored,and there was an elevation in the expression levels of cuproptosis-related genes,lipDLAT,lipDBT,lipDLST,thioredoxin,and LA mRNA(P＜0.05).Conversely,the expression levels of DLAT dimer decreased(P＜0.05).Compared with the penicillamine group,the levels of ALT and AST decreased in the Gandou Decoction combined with penicillamine group(P＜0.05),the copper content in liver tissue decreased(P＜0.01),the mRNA expression of PDHB,iron-sulfur cluster protein and lipoylated protein increased(P＜0.01,P＜0.05),and the expressions of DLAT dimer and HSP70 decreased(P＜0.01).Conclusions The combination of Gandou Decoction and penicillamine demonstrates enhanced copper excretion and increased expression of LA pathway-related proteins,leading to the mitigation of hepatic cuproptosis in a WD model.Furthermore,Gandou Decoction exhibits an auxiliary potentiating effect when used in conjunction with penicillamine for the treatment of WD.
Wilson diseaseCuproptosisGandou Decoctionpenicillamineintegrative medicineChinese herbal compound
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