Clinical Significance Of Circ-0007385/miR-181b/notch-1 In Lupus Nephritis

Purpose:To detect the expression level of circ＿0007385 in Peripheral Blood Mononuclear Cell(PBMC)of patients with Lupus nephritis(LN),and analyze the clinical significance.To explore the possible mechanism of circ＿0007385 as ceRNA regulating miRNA-181b and activating notch-1 signal to participate in the pathogenesis of lupus nephritis.Methods:Sixty patients with SLE were randomly divided into two groups,including LN group and N-LN group,and 30 healthy subjects were selected as control group(HC group).Screening three circrnas(circ-0002416,circ-0000387,circ-0007385)whose expression levels of circleRNAs in LN patients’ PBMC were more than 2 times different by high-throughput sequencing in the previous stage for clinical verification.The up-regulated expression of circ-0007385 was confirmed by RT-qPCR with statistical significance.TargetScan and miRanda software analysis confirmed that circ-0007385 binds to miR-181b relatively stably,and Notch-1 mrna is one of the potential targets of miR-181b.The selected circ-0007385,miR-181b and Notch-1 mRNA were independently verified by RT-qPCR,and the notch signaling ligand Jagged-1 concentration was detected by ELISA.The sensitivity and specificity of circ-0007385 and miR-181b for the diagnosis of LN patients were analyzed by ROC curve.Results:1.The expression level of circ-0007385 in PBMC of SLE group was significantly lower than that in HC group(p<0.01),and the expression level of circ-0007385 was positively correlated with anti-DSDNA antibody(r=0.6192 p<0.001).It was positively correlated with SLEDAI-2K score(r=0.6045,p<0.01)and negatively correlated with complement C3(r=-0.4762,p<0.05).The expression level of circ-0007385 in PBMC of LN group was significantly lower than that of N-LN group(p<0.01),and the expression level of circ-0007385 was positively correlated with 24h urinary protein quantization(r=0.3864,p<0.01).2.The expression level of miR-181b in PBMC of SLE group was significantly higher than that of HC group(p<0.01),and the expression level of miR-181b in PBMC of LN group was significantly higher than that of N-LN group(p<0.01).It was negatively correlated with the expression of circ-0007385(r=-0.6423,p<0.05).3.ROC curve analysis showed that the AUC of circ-0007385 combined with miR-181b in diagnosing LN was 0.79,p<0.01,higher than that of circ＿0007385(AUC=0.77,p<0.01)and miR-181b(AUC=0.68,p<0.05).4.Compared with the N-LN group,Notch-1 mRNA expression level in the PBMC of the LN group was significantly increased(p<0.01),and was negatively correlated with the relative expression level of miR-181b(r=-0.4260,p<0.05).Serum Jagged-1 protein level was significantly increased(p<0.01).Conclusion:circ＿0007385 in human PBMC may be involved in the pathogenesis of lupus nephritis as ceRNA competitively binds miRNA-181b and activates notch-1 signal.