Simiao Yong’an Decoction Alleviates Thromboangiitis Obliterans By Regulating MiR-548j-5p/IL-17A Signaling Pathway

Purpose:From the perspective of epigenetics,this paper discusses the role of micro RNA and interleukin-17A(IL-17A)in the occurrence of Thromboangiitis obliterans(TAO)and the targeted regulation mechanism of Simiao Yong ’an Decoction,so as to provide new targets and approaches for clinical diagnosis and treatment.Method:Peripheral blood mononuclear cells(PBMCs)of normal people and patients with TAO were collected,and the m RNA expressions of miR-548j-5p and IL-17 A m RNA PBMCs of patients with TAO were detected by quantitative real-time polymerase chain reaction(q RT-PCR).Enzyme linked immunosorbent assay(ELISA)was used to detect the level of IL-17 A protein in PBMCs.Mi RNA microarray kit was used to detect the whole transcriptome of RNAs extracted from PBMCs,and combined with bioinformatics analysis to screen mirnas regulated upstream of IL-17 A.To regulate the expression of miR-548j-5p in human umbilical vein endothelial cells(HUVECs)and observe its effect on IL-17 A.The interaction between miR-548j-5p and IL-17 A m RNA was detected by double luciferase reporter system.TAO rat model was established,and the effects of Simiao Yong ’an Decoction on miR-548j-5p and IL-17 A were observed in vivo and in vitro.At the same time,I injected IL-17 A antagonist into the abdominal cavity of TAO rats to observe the changes of thrombosis,and after using miR-548j-5p inhibitor,I observed the therapeutic effect of Simiao Yong ’an Decoction in vivo and in vitro.To analyze the therapeutic effect and molecular mechanism of Simiao Yong ’an Decoction /miR-548j-5p/IL-17 A signal axis on TAO.Results:1.the results of q Rt-PCR and ELISA showed that the expression of IL-17 A in PBMCs of patients with TAO was significantly higher than that of normal group(P< 0.05),suggesting that IL-17 A was involved in the occurrence of TAO.2.The results of high-throughput microarray detection combined with Target Scan and miRDB analysis showed that the expression of miR-548j-5p in TAO patients was decreased,and there was a potential binding site with IL-17 A.q RT-PCR verified that the expression of miR-548j-5p in PBMCs of TAO patients was significantly decreased,which was statistically significant compared with that of normal controls(P< 0.05).Pearson correlation analysis showed that miR-548j-5p was significantly decreased.P< 0.05),ROC analysis showed that the expression of miR-548j-5p could sensitively distinguish patients with TAO from normal controls.3.The double luciferase reporter gene confirmed that miR-548j-5p directly combined with the 3’UTR of IL-17 A m RNA through base complementation.4.Up-regulating the expression of miR-548j-5p in HUVECs can significantly inhibit the expression of IL-17 A,TF and v WF(P< 0.05),and promote the expression of e NOS(P< 0.05),while down-regulating miR-548j-5p has the opposite expression(P< 0.05).5.After intraperitoneal injection of Anti-IL-17 A into TAO rats,the thrombus area decreased significantly(P< 0.05).6.Serum containing drug(Simiao Yong ’an Decoction)in rats can up-regulate the expression of miR-548j-5p,inhibit the expression of IL-17 A,TF and v WF(P< 0.05),and promote the expression of e NOS(P< 0.05).After inhibiting the expression of miR-548j-5p,the therapeutic effect of Simiao Yong ’an Decoction decreased significantly(P< 0.05).7.After the gastric administration of Simiao Yong ’an Decoction,the thrombus in the femoral artery of TAO rats decreased,the expression of miR-548j-5p in PBMCs increased,the expressions of IL-17 A,TF and v WF decreased(P< 0.05),and the expression of e NOS increased(P< 0.05).8.Injecting miR-548j-5p inhibitor into 8.TAO rats can reverse the inhibitory effect of Simiao Yong ’an Decoction on the expression of IL-17 A,TF and v WF(P< 0.05)and promote the expression of e NOS(P< 0.05).Conclusion:1.The expression of miR-548j-5p decreased and the expression of IL-17 A increased,which participated in the occurrence of TAO;2.miR-548j-5p directly binds to IL-17 A m RNA 3’UTR through base complementation,thus inhibiting its expression;3.Simiao Yong ’an Decoction has therapeutic effect on TAO by regulating the signal axis of miR-548j-5p/IL-17 A.