Clinical Efficacy And Ferroptosis Related Indicators Of Gandouling In The Treatment Of Wilson Disease With Phlegm-Blood Stasis

1 ObjectivesIdentification of ferroptosis key genes in the diagnosis and treatment of Wilson disease(WD)by bioinformatics analysis and machine learning,and to observe the effects of Gandouling on clinical efficacy,TCM syndrome score,ferroptosis-related indicators and oxidative stress-related indicators in patients of WD with phlegm-blood stasis.2 Methods2.1 Bioinformatics and machine learning researchThe eligible raw dataset(GSE107323)was retrieved from the Gene Expression Database(GEO),normalized,and screened for the differentially expressed genes(DEGs)of WD.Ferroptosis-related genes(FRGs)were downloaded from the Ferr Db database,and the differential FRGs of WD were obtained after taking the intersection of the two,and enrichment analyses were conducted.Subsequently,ferroptosis key genes that could be used for the diagnosis and treatment of WD were further identified by machine learning algorithms.2.2 Clinical researchThe 124 patients of WD with phlegm-blood stasis were collected from January2022 to December 2022 in the Department of Encephalopathy of the First Affiliated Hospital of Anhui University of Chinese Medicine,and they were divided into observation group and control group,with 62 patients in each group.The control group was treated with conventional copper excretion therapy.The observation group was treated with Gandouling on the basis of the treatment in the control group.The clinical efficacy,TCM syndrome score,as well as the level of ferroptosis-related indicators such as glutathione peroxidase 4(GPX4),ceruloplasmin(CER),transferrin(TF),and oxidative stress-related indicators such as glutathione peroxidase(GSH-Px),superoxide dismutase(SOD),malondialdehyde(MDA)of the two groups before and after treatment were observed.3 Results3.1 Bioinformatics and machine learning part(1)A total of 3393 DEGs of WD were identified in the GSE107323 dataset,and484 FRGs were downloaded from the Ferr Db database,and 111 differential FRGs of WD were obtained by taking the intersection of the two.(2)Gene ontology(GO)enrichment results showed that the biological processes(BP)of differential FRGs of WD mainly involved responses to oxidative stress and responses to nutrient levels,etc.Cellular components(CC)mainly involved apical part of cells and basolateral plasma membrane,etc.Molecular functions(MF)mainly involved iron ion binding and ubiquitin-like protein ligase binding,etc.The results of Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis showed that differential FRGs of WD were mainly enriched in 11 signaling pathways including ferroptosis,peroxisome proliferator-activated receptor(PPAR)and fatty acid metabolism,etc.(3)Four key ferroptosis genes that could be used in the diagnosis and treatment of WD were further identified by machine learning algorithms,namely GPX4,TF,CARS1 and SIRT3.3.2 Clinical parts(1)Compared with the control group(62.90%,39/62),the total effective rate of the observation group(82.26%,51/62)was significantly higher(P < 0.05).(2)The difference between the TCM syndrome scores of patients in the two groups before treatment was not statistically significant(P > 0.05).Compared with before treatment,the TCM syndrome scores of both groups decreased significantly after treatment(P<0.01);Compared between the groups after treatment,the decrease was more significant in the observation group than in the control group(P<0.01).(3)The differences in ferroptosis-related indicators between the two groups before treatment were not statistically significant(P>0.05).Compared with before treatment,the TF and GPX4 levels were significantly higher in both groups after treatment(P<0.01),and there were no statistical differences in CER levels(P > 0.05);Compared between the groups after treatment,the TF and GPX4 levels were more significantly higher in the observation group than in the control group(P<0.01),and there were no statistical differences in CER levels(P>0.05).(4)The differences in oxidative stress-related indicators between the two groups before treatment were not statistically significant(P > 0.05).Compared with before treatment,the GSH-Px and SOD levels were significantly higher(P<0.01),and the MDA levels were significantly lower(P < 0.01)in both groups after treatment.Compared between the groups after treatment,the GSH-Px and SOD levels were more significantly higher in the observation group than in the control group(P < 0.01),and MDA levels were significantly lower(P < 0.01).4 Conclusion(1)GPX4,TF,CARS1 and SIRT3 may be the key ferroptosis genes in the diagnosis and treatment of WD.(2)Gandouling has unique advantages in improving the clinical efficacy and reducing the TCM syndrome score in patients of WD with with phlegm-blood stasis,and significantly improves the levels of indicators related to oxidative stress and ferroptosis.