Abstract：
Thromboangiitis obliterans (TAO) is a rare, chronic, progressive, and segmental inflammatory disease characterized by a high rate of amputation, significantly compromising the quality of life of patients. Si-Miao-Yong-An decoction (SMYA), a traditional prescription, exhibits anti-inflammatory, anti-thrombotic, and various other pharmacological properties. Clinically, it was fully proved to be effective for TAO therapy, but the specific therapeutic effect of SMYA on TAO has been unknown. Thus, deep unveiling the mechanism of SMYA in TAO for identifying clinical therapeutic targets is extremely important. In this study, we observed elevated levels of IL-17A in the peripheral blood mononuclear cells (PBMCs) of TAO patients, whereas the expression of miR-548j-5p was significantly decreased. A negative correlation between the levels of miR-548j-5p and IL-17A was also demonstrated. In vitro experiments showed that overexpression of miR-548j-5p led to a decrease in IL-17A levels, whereas downregulation of miR-548j-5p showed the opposite effect. Using a dual luciferase assay, we confirmed that miR-548j-5p directly targets IL-17A. Furthermore, serum containing SMYA effectively decreased IL-17A levels by increasing the expression of miR-548j-5p. More importantly, the results of in vivo tests indicated that SMYA mitigated the development of TAO by inhibiting IL-17A through the upregulation of miR-548j-5p in vascular tissues. In conclusion, SMYA significantly enhances the expression of miR-548j-5p, thereby reducing the levels of the target gene IL-17A and alleviating TAO. Our research not only identifies novel targets and pathways for the clinical diagnosis and treatment of TAO but also advances the innovation in traditional Chinese medicine through the elucidation of the SMYA/miR-548j-5p/IL-17A regulatory axis in the pathogenesis of TAO.
摘要：
血栓闭塞性脉管炎（TAO）是一种罕见的，慢性，进步,以高截肢率为特征的节段性炎症性疾病，显著影响患者的生活质量。四苗勇安汤（SMYA），传统的处方,表现出抗炎，抗血栓,和其他各种药理特性。临床上,它被充分证明对TAO治疗是有效的，但SMYA对TAO的具体治疗效果尚不清楚。因此，深入揭示SMYA在TAO中的作用机制对于识别临床治疗靶点极为重要。在这项研究中,我们观察到TAO患者外周血单核细胞（PBMC）中IL-17A的水平升高，而miR-548j-5p的表达显著降低。还证明了miR-548j-5p和IL-17A的水平之间的负相关。体外实验表明，miR-548j-5p的过表达导致IL-17A水平降低，而miR-548j-5p的下调显示出相反的效果。使用双荧光素酶测定法，我们证实miR-548j-5p直接靶向IL-17A。此外，含有SMYA的血清通过增加miR-548j-5p的表达有效降低IL-17A水平。更重要的是,体内试验结果表明,SMYA通过上调血管组织中miR-548j-5p抑制IL-17A,从而减轻TAO的发育.总之,SMYA显著增强miR-548j-5p的表达,从而降低靶基因IL-17A的水平并减轻TAO。我们的研究不仅为TAO的临床诊断和治疗确定了新的靶点和途径，而且通过阐明SMYA/miR-548j-5p/IL-17A调控轴在TAO发病机制中的作用，推进了中医药的创新。
