Effects Of Clozapine Regulating MiRNAs On Abnormal Lipid Metabolism In Patients With Schizophrenia

Objective(s):To explore the changes of miRNAs and their possible regulatory mechanisms in the process of clozapine treatment on blood lipid metabolism in schizophrenics.Methods:1.Select 6 samples(3 patients with dyslipidemia in schizophrenia comorbidities treated with clozapine alone from the case group,and 3 patients from the control group),take their fasting blood,and use chip screening to detect the expression of miRNAs,The Kyoto Encyclopedia of Genes and Genomes(KEGG)and gene ontology(GO)enrichment analysis were performed on miRNAs with significant differences.2.Perform bioinformatics analysis,predict target genes for differentially expressed miRNAs,and search for genes related to lipid metabolism abnormalities.Use the independently developed software miRMATCHV1.0 to perform data matching screening to find overlapping genes.Based on the analysis results and previous studies,carry out research on ADIPOQ.3.The differential miRNA expression in venous blood of 47 samples(12 in the control group and 35 in the case group)was detected by qPCR to verify the results of the pre chip screening.4.The expression of differential miRNA in the liver of schizophrenic mice after clozapine intervention was detected by qPCR(n=3).5.Immunohistochemistry(IHC)and Western blot(WB)were used to detect the expression of ADIPOQ in the liver of schizophrenic mice after clozapine intervention(n=3).6.The luciferase reporter gene detection experiment clarified the regulatory relationship between differential miRNA and ADIPOQ(n=3).Results:1.Chip screening detected 25 differentially expressed miRNAs:miR-485-5p,novel268_mature、miR-133a-3p、miR-543、miR-205-5p、miR-29b-3p、novel329_mature、miR-320c、miR-375-3p、miR-381-3p、miR-4446-3p、miR-4731-5p、miR-873-5p、miR-654-3p、miR-29c-3p、novel281_Nature,miR-504-5p,and miR-1-3p increase,novel397_Material,miR-423-5p,miR-1224-5p,miR-16-2-3p,miR-92b-5p,miR-4732-5p,and miR-5189-5p decreased.2.According to bioinformatics analysis,ADIPOQ may be one of the target genes regulated by miR-485-5p.3.Experimental results of qPCR in human blood samples:Compared with the control group,there was no statistically significant difference in the expression of miR-485-5P in the group with abnormal lipid metabolism(P>0.05).The expression of miR-485-5P in the schizophrenia comorbidity group treated with clozapine alone was significantly higher than that in the schizophrenia group treated with clozapine alone(P<0.05).There was no statistically significant difference in the expression of miR-485-5P between the schizophrenia group treated with clozapine alone and the schizophrenic comorbidities with abnormal lipid metabolism group treated with clozapine alone compared with the untreated schizophrenic group(P>0.05).4.Experimental results of qPCR in mouse liver:The expression of miR-485-5P in schizophrenic mice treated with clozapine was significantly higher than that in schizophrenic mice(P<0.05).5.Immunohistochemical test results:ADIPOQ was expressed in the cytoplasm of the mouse liver.6.Western blot analysis showed that the expression of ADIPOQ in schizophrenic mice treated with clozapine was significantly lower than that in control mice(P<0.05);The expression of ADIPOQ in schizophrenic mice treated with clozapine was significantly lower than that in schizophrenic mice(P<0.05).7.Experimental results of luciferase reporter gene detection:No targeted regulatory relationship between miR-485-5P and ADIPOQ was found.Conclusion(s):1.Clozapine may mediate abnormal lipid metabolism in patients with schizophrenia by influencing changes in miRNAs.2.MiR-485-5p may be related to the effect of clozapine on lipid metabolism.3.Clozapine may indirectly decrease ADIPOQ by upregulating miR-485-5p,leading to abnormal lipid metabolism.However,the regulatory relationship between the two target genes is unclear.