Inhibition of miR-155-5p attenuates the valvular damage induced by rheumatic heart disease
Inhibition of miR-155-5p attenuates the valvular damage induced by rheumatic heart disease
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DOI：
10.3892/ijmm.2019.4420
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发表时间：
2020-02-01
影响因子：
5.4
通讯作者：
Zeng, Zhiyu
中科院分区：
文献类型：
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作者：
Chen, Ang;Wen, Jianlin;Zeng, Zhiyu
自身免疫参与了风湿性心脏病（RHD）导致的瓣膜损伤。越来越多的证据表明，微小核糖核酸（miRNA/miR）与自身免疫性疾病相关。信号转导及转录激活因子3（STAT3）、1 - 磷酸鞘氨醇受体1（S1PR1）和细胞因子信号传导抑制因子1（SOCS1）在自身免疫和炎症中的作用已得到广泛研究。因此，本研究旨在探究miR - 155 - 5p通过S1PR1、SOCS1/STAT3以及白细胞介素（IL）- 6/STAT3信号通路在RHD诱导的瓣膜损伤中所起的作用。通过灭活的A组链球菌和完全弗氏佐剂诱导建立RHD大鼠模型。使用重组腺相关病毒（AAV - miR155 - 抑制剂）抑制心脏中miR - 155 - 5p的表达。通过苏木精 - 伊红染色和天狼星红染色评估炎症和纤维化情况。采用逆转录 - 定量聚合酶链反应检测瓣膜组织和血清外泌体中miR - 155 - 5p的表达。通过蛋白质免疫印迹法和免疫组织化学法检测S1PR1、SOCS1、STAT3、磷酸化STAT3、IL - 6和IL - 17的蛋白表达。通过双荧光素酶报告基因实验检测miR - 155 - 5p与S1PR1及SOCS1之间的关系。采用酶联免疫吸附试验检测细胞因子浓度。在RHD模型中，瓣膜组织和血清外泌体中miR - 155 - 5p的表达增加，同时S1PR1表达降低，SOCS1/STAT3信号通路激活。瓣膜和血清中IL - 6和IL - 17的表达增加。双荧光素酶报告基因实验表明，miR - 155 - 5p直接作用于S1PR1和SOCS1。通过AAV预处理抑制瓣膜miR - 155 - 5p可增加S1PR1表达，抑制SOCS1/STAT3信号通路的激活，从而减轻瓣膜炎症和纤维化，同时降低瓣膜和血清中IL - 6和IL - 17的水平。这些结果表明，抑制miR - 155 - 5p可通过S1PR1、SOCS1/STAT3和IL - 6/STAT3信号通路减轻RHD诱导的瓣膜损伤。
Autoimmunity is involved in the valvular damage caused by rheumatic heart disease (RHD). Increased evidence has linked microRNAs (miRNAs/miRs) to autoimmune disease. Signal transducer and activator of transcription 3 (STAT3) and sphingosine-1-phosphate receptor 1 (S1PR1) and suppressor of cytokine signaling 1 (SOCS1) have been widely studied for their roles in autoimmunity and inflammation. Thus, the current study aims to investigate the role played by miR-155-5p in RHD-induced valvular damage via the S1PR1, SOCS1/STAT3 and interleukin (IL)-6/STAT3 signaling pathways. An RHD rat model was induced by inactivated Group A streptococci and complete Freund's adjuvant. A recombinant adeno-associated virus (AAV-miR155-inhibitor) was used to inhibit the expression of miR-155-5p in the heart. Inflammation and fibrosis were assessed by hematoxylin and eosin staining and Sirius red staining. The expression of miR-155-5p in valvular tissues and serum exosomes was detected by reverse transcription-quantitative PCR. S1PR1, SOCS1, STAT3, phosphorylated STAT3, IL-6 and IL-17 protein expression was detected by western blotting and immunohistochemistry. The relationships between miR-155-5p and S1PR1 and SOCS1 were detected by dual luciferase assays. Cytokine concentrations were measured by ELISA. The expression of miR-155-5p in valve tissues and serum exosomes was increased along with decreased S1PR1 and activated SOCS1/STAT3 signaling in the RHD model. The expression of IL-6 and IL-17 was increased in the valves and the serum. Dual luciferase assays showed that miR-155-5p directly targeted S1PR1 and SOCS1. Inhibition of valvular miR-155-5p through AAV pretreatment increased S1PR1 expression and inhibited activation of the SOCS1/STAT3 signal pathway as a result of attenuated valvular inflammation and fibrosis as well as a decrease in IL-6 and IL-17 in the valves and serum. These results suggest that inhibition of miR-155-5p can reduce RHD-induced valvular damage via the S1PR1, SOCS1/STAT3 and IL-6/STAT3 signaling pathways.