| Background Systemic lupus erythematosus（SLE）is a multisystem autoimmune disease characterized by the production of antinuclear antibody（ANA）and its immune complexes.At present,the mechanism of SLE is not completely clear.In recent years,some studies have confirmed that exosomes can participate in the pathogenesis of SLE through their inclusions.ObjectiveTo explore the differential expression of long non-coding RNAs（lnc RNAs）in peripheral blood exosomes of SLE patients,and to explore its clinical significance in the diagnosis and treatment of SLE.MethodsFifty-two SLE patients admitted to our hospital from August 2019 to August 2021 were selected,including 26 SLE active group and 26 SLE stable group.The clinical data of each patient was recorded and the SLE disease activity Index（SLEDAI）score was evaluated.At the same time,20 rheumatoid arthritis（RA）patients were selected as case controls and 26 healthy volunteers as healthy controls with matched gender and age.Peripheral venous blood of all subjects was extracted,and the morphology and particle size of isolated plasma exosomes were identified by projection electron microscopy and particle size analyzer to explore the expression of lnc RNAs in exosomes.First,high-throughput sequencing technology was used to detect the expression of plasma exosome lnc RNAs in patients with active SLE（6 cases）,stable SLE（6 cases）and normal control（6 cases）.The expression profile of plasma exosome lnc RNAs in SLE patients was constructed to screen for differentially expressed lnc RNAs.Secondly,q RT-PCR was used to verify the independent samples of 5 candidate differential lnc RNAs screened by sequencing（20 cases of active SLE,20 cases of stable SLE,20 cases of normal control and 20 cases of RA disease control）,and predict the target genes and target gene functions of the differentially expressed lnc RNAs.ROC curve was used to analyze the diagnostic value of differentially expressed lnc RNAs in plasma exosomes for SLE,and the relationship between them and SLE disease activity（SLEDAI）was analyzed.Results1.Under projective electron microscopy,exosomes needed for research were successfully obtained,which were circular vesicles with particle sizes ranging from 30 nm to 150 nm.2.By high-throughput sequencing,8,836 genes were up-regulated and 5,914 genes were down-regulated in the SLE group compared with the healthy control group.Combined with relevant references,lnc RNAs that may be involved in immune and inflammatory pathways were selected,and exosomal lnc RNAs with large gene differences and high expression volume were selected according to sequencing results.Five exosomal lnc RNAs（up-regulated NEAT1,LINC00667,PSMA3-AS1,DANCR and down-regulated TPT1-AS1）were screened for SLE disease.The PI3K/Akt signaling pathway,Rap1 signaling pathway and Ras signaling pathway may be involved（Figure 3.8）.3.There were no significant differences in the relative expression of exosome NEAT1 between SLE group and healthy control group or between active SLE group and stable SLE group（P > 0.05）.4.The expression level of exosome LINC00667 in SLE group was higher than that in healthy control group（P < 0.05）.ROC curve showed that the AUC of LINC00667 relative expression level for the diagnosis of SLE was 0.815（95%CI: 0.704～0.926,P < 0.05）,with a sensitivity of 80% and specificity of 75%.The relative expression level of LINC00667 was positively correlated with SLEDAI（P < 0.05）,but there was no significant difference in the expression level of LINC00667 in exosomes between SLE active group and SLE stable group（P > 0.05）.5.The expression level of PSMA3-AS1 in SLE group was higher than that in healthy control group（P < 0.05）.ROC curve showed that the AUC of PSMA3-AS1 relative expression level for the diagnosis of SLE was 0.892（95%CI: 0.806-0.978,P < 0.05）,with a sensitivity of 95%and specificity of 75%.There was no significant difference in PSMA3-AS1 relative expression between SLE active group and SLE stable group（P > 0.05）.6.The expression level of exosome DANCR in SLE group was higher than that in healthy control group（P < 0.05）.ROC curve showed that the AUC of DANCR relative expression level for the diagnosis of SLE was 0.759（95%CI: 0.629-0.888,P < 0.05）,with a sensitivity of85% and specificity of 70%.The expression of DANCR in SLE stable group was lower than that in SLE active group（P < 0.05）.The relative expression level of DANCR was positively correlated with SLEDAI（P < 0.05）.7.The relative expression levels of LINC00667,PSMA3-AS1 and DANCR for the combined diagnosis of SLE were AUC=0.910（95%CI: 0.831-0.989,P < 0.05）,with sensitivity of 95%and specificity of 75%.8.There was no significant difference in the expression level of TPT1-AS1 in SLE group compared with healthy control group（P<0.05）.The expression level in SLE stable group was higher than that in SLE active group（P < 0.05）.The relative expression level of TPT1-AS1 was negatively correlated with CRP（P < 0.05）.9.The relative expression levels of LINC00667,PSMA3-AS1,DANCR and TPT1-AS1 in SLE group were higher than those in RA group（P < 0.05）.10.There were no significant differences in the relative expression levels of NEAT1,LINC00667,PSMA3-AS1,DANCR and TPT1-AS1 between the groups with abnormal renal function and normal renal function or with and without proteinuria（P > 0.05）.Conclusions1.Compared with healthy controls,differentially expressed LINC00667,PSMA3-AS1 and DANCR in plasma exosomes of SLE patients have potential diagnostic value for SLE,and combined detection has greater potential diagnostic value.2.Differentially expressed DANCR and TPT1-AS1 may be used as reference biomarkers for clinical evaluation of SLE disease activity.3.Differentially expressed lnc RNAs in plasma exosomes of SLE patients may involve the PI3K/Akt signaling pathway,Rap1 signaling pathway and Ras signaling pathway.Further exploration of their detailed mechanism of action may find new potential targets for the treatment of SLE. |
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