MiR-223-3p Alleviates Vascular Endothelial Injury by Targeting IL6ST in Kawasaki Disease
MiR-223-3p Alleviates Vascular Endothelial Injury by Targeting IL6ST in Kawasaki Disease
复制标题
MiR-223-3p 通过靶向 IL6ST 减轻川崎病中的血管内皮损伤
DOI：
10.3389/fped.2019.00288
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发表时间：
2019-07-24
影响因子：
2.6
通讯作者：
Lv, Hai tao
中科院分区：
文献类型：
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作者：
Wang, Xiang;Ding, Yue yue;Lv, Hai tao
背景：川崎病（KD）是一种以急性系统性血管炎症为特征的自限性疾病。它会引起大多数中小型动脉的病理变化，尤其是冠状动脉，这增加了成人患冠心病的风险。材料和方法：我们使用 miRNA 微阵列和 RT-PCR 检测了 30 名 KD 患者和 12 名正常对照中 miR-223-3p 的表达。采用白色念珠菌水不溶物(CAWS)构建KD小鼠模型。我们还使用 qRT-PCR 检查了 miR-223-3p 的表达。采用荧光素酶报告系统来验证 miR-223-3p 和白细胞介素 6 受体亚基 β (IL-6ST) 之间的相关性。使用TNF-α刺激人冠状动脉内皮细胞（HCAEC），并使用miR-223-3p激活剂或抑制剂以及KD血清来治疗HCAEC。采用Western blotting自动定量分析蛋白印迹系统检测信号转导和转录激活因子3（STAT3）、磷酸化信号转导和转录激活因子3（pSTAT3）和NF-κB p65的表达。结果：临床试验发现，急性川崎病组与对照组的miR-223-3p表达存在显着差异（2倍以上）。 KD 患者中 E-选择素和细胞间细胞粘附分子 1 (ICAM-1) 水平也显着升高（约 2 倍），尤其是冠状动脉病变时。 MiR-223-3p可减轻KD小鼠血管内皮损伤，IL-6（白细胞介素6）、E-选择素和ICAM-1同时阴性。激动剂治疗小鼠和KD小鼠之间IL-6、E-选择素和ICAM-1 mRNA表达值降低，而IL-6ST值升高。 RT-qPCR 结果显示，miR-223-3p 在 KD 小鼠和激动剂组中均在第三天达到最高表达。 MiR-223-3p可以直接与IL-6ST 3'非翻译区（UTR）结合并抑制IL-6的表达。 miR-223的过表达下调了IL6ST的表达，并降低了p-STAT3和NF-κB p65的表达，而miR-223抑制剂可以逆转上述过程。结论：MiR-223-3p是川崎病血管内皮损伤的重要调控因子，有可能成为未来川崎病治疗的潜在靶点。
Background: Kawasaki disease (KD) is a self-limiting illness with acute systematic vascular inflammation. It causes pathological changes in mostly medium and small-sized arteries, especially the arteria coronaria, which adds the risk of developing coronary heart disease in adults. Materials and methods: We detected the miR-223-3p expression in 30 KD patients combined with 12 normal controls using miRNA microarrays and RT-PCR. A KD mouse model was constructed using Candida albicans water insoluble substance (CAWS). We also checked the miR-223-3p's expression using qRT-PCR. The Luciferase reporting system was implemented to validate the correlation between miR-223-3p and Interleukin-6 receptor subunit beta (IL-6ST). TNF-α was used to stimulate human coronary artery endothelial cells (HCAECs), and miR-223-3p activator or inhibitor and KD serum were used to treat HCAECs. A Western blotting automatic quantitative analysis protein imprinting system was used to test the expression of signal transducer and the activator of transcription 3 (STAT3), phosphorylated-signal transducer and the activator of transcription 3 (pSTAT3) and NF-κB p65. Results: Clinical trials found that miR-223-3p expressions were markedly different (more than 2-fold) between the acute KD group and the control group. E-selectin and intercellular cell adhesion molecule-1 (ICAM-1) levels were also significantly higher (about 2-fold) in KD especially with coronary artery lesions. MiR-223-3p could alleviate vascular endothelial damage in KD mice, and IL-6 (Interleukin-6), E-selectin and ICAM-1 were simultaneously negative. The values of IL-6, E-selectin, and ICAM-1 mRNA expressions decreased, while the value of IL-6ST was increased between the agonist treated mice and KD mice. The RT-qPCR consequences displayed that miR-223-3p explored the highest expression on the third day in both the KD mice as well as the agonist group. MiR-223-3p can directly combine with IL-6ST 3' untranslatable regions (UTR) and held back the IL-6's expression. Overexpression of miR-223 down regulated IL6ST expression and decreased the expression of p-STAT3 and NF-κB p65, while the miR-223 inhibitor could reverse the above process. Conclusion: MiR-223-3p is an important regulatory factor of vascular endothelial damage in KD and could possibly become a potential target of KD treatment in the future.