Lu Qiyu, Sun Yi, Duan Yuyin, Li Bin, Xia Jianming, Yu Songhua, Zhang Guimin
Department of Gastrointestinal Surgery, The Second Affiliated Hospital of Kunming Medical University, Kunming, Yunnan Province, 650101, China.
Department of Cardiothoracic Surgery, The First Affiliated Hospital of Kunming Medical University, Kunming, Yunnan Province, 650101, China.
BMC Cardiovasc Disord. 2018 Mar 16;18(1):53. doi: 10.1186/s12872-018-0788-2.
Valvular heart disease is a leading cause of cardiovascular mortality, especially in China. More than a half of valvular heart diseases are caused by acute rheumatic fever. microRNA is involved in many physiological and pathological processes. However, the miRNA profile of the rheumatic valvular heart disease is unknown. This research is to discuss microRNAs and their target gene pathways involved in rheumatic heart valve disease.
Serum miRNA from one healthy individual and four rheumatic heart disease patients were sequenced. Specific differentially expressed miRNAs were quantified by Q-PCR in 40 patients, with 20 low-to-moderate rheumatic mitral valve stenosis patients and 20 severe mitral valve stenosis patients. The target relationship between certain miRNA and predicted target genes were analysis by Luciferase reporter assay. The IL-1β and IL1R1 expression levels were analyzed by immunohistochemistry and western blot in the mitral valve from surgery of mitral valve replacement.
The results showed that 13 and 91 miRNAs were commonly upregulated or downregulated in all four patients. Nine miRNAs, 1 upregulated and 8 downregulated, that had a similar fold change in all 4 patients were selected for quantitative PCR verification. The results showed similar results from miRNA sequencing. Within these 9 tested miRNAs, hsa-miR-205-3p and hsa-miR-3909 showed a low degree of dispersion between the members of each group. Hsa miR-205-3p and hsa-miR-3909 were predicted to target the 3'UTR of IL-1β and IL1R1 respectively. This was verified by luciferase reporter assays. Immunohistochemistry and Western blot results showed that the mitral valve from rheumatic valve heart disease showed higher levels of IL- 1β and IL1R1 expression compared with congenital heart valve disease. This suggested a difference between rheumatic heart valve disease and other types of heart valve diseases, with more inflammatory responses in the former.
In the present study, by next generation sequencing of miRNAs, it was revealed that interleukin 1β and interleukin 1 receptor 1 was involved in rheumatic heart diseases. And this is useful for diagnosis and understanding of mechanism of rheumatic heart disease.
心脏瓣膜病是心血管疾病死亡的主要原因之一，在中国尤其如此。超过一半的心脏瓣膜病由急性风湿热引起。微小RNA参与许多生理和病理过程。然而，风湿性心脏瓣膜病的微小RNA图谱尚不清楚。本研究旨在探讨参与风湿性心脏瓣膜病的微小RNA及其靶基因途径。
对1名健康个体和4名风湿性心脏病患者的血清微小RNA进行测序。通过Q-PCR对40例患者（20例轻至中度风湿性二尖瓣狭窄患者和20例重度二尖瓣狭窄患者）中特定差异表达的微小RNA进行定量分析。通过荧光素酶报告基因检测分析特定微小RNA与预测靶基因之间的靶向关系。通过免疫组织化学和蛋白质印迹法分析二尖瓣置换手术中二尖瓣组织中白细胞介素-1β（IL-1β）和白细胞介素-1受体1（IL1R1）的表达水平。
结果显示，在所有4例患者中，分别有13种和91种微小RNA普遍上调或下调。选择在所有4例患者中具有相似倍数变化的9种微小RNA（1种上调和8种下调）进行定量PCR验证。定量PCR结果与微小RNA测序结果相似。在这9种检测的微小RNA中，hsa-miR-205-3p和hsa-miR-3909在每组成员之间显示出较低的离散度。预测hsa-miR-205-3p和hsa-miR-3909分别靶向IL-1β和IL1R1的3'非翻译区（3'UTR）。荧光素酶报告基因检测验证了这一结果。免疫组织化学和蛋白质印迹结果显示，与先天性心脏瓣膜病相比，风湿性心脏瓣膜病患者的二尖瓣组织中IL-1β和IL1R1表达水平更高。这表明风湿性心脏瓣膜病与其他类型的心脏瓣膜病存在差异，前者炎症反应更明显。
在本研究中，通过对微小RNA进行二代测序，发现白细胞介素-1β和白细胞介素-1受体1参与了风湿性心脏病的发生。这对于风湿性心脏病的诊断和发病机制的理解具有重要意义。
