Shan Haiyan, Guo Dawei, Zhang Siyang, Qi Huimeng, Liu Shen, Du Yanmei, He Yini, Wang Bofu, Xu Ming, Yu Xiaosong
Department of General Practice, The First Affiliated Hospital of China Medical University, No. 155 Nanjing North Street, Heping District, Shenyang, 110001, China.
Department of the Fourth General Surgery, The Fourth Affiliated Hospital of China Medical University, No. 4 Chongshan East Road, Huanggu District, 110032, Shenyang, China.
Cell Biosci. 2020 Jan 7;10:4. doi: 10.1186/s13578-019-0371-2. eCollection 2020.
Plenty of long non-coding RNAs (lncRNAs) play vital roles in the progression of atherosclerosis. Small nucleolar RNA host gene 6 (SNHG6) is a well known lncRNA that is aberrantly high expressed in atherosclerosis patients. However, its function and basic mechanism in atherosclerosis events have not been well clarified.
The expression patterns of SNHG6, miR-135a-5p, ROCK1 and ROCK2 in clinical samples and cells were detected by RT-qPCR assays. Cell Counting Kit-8 (CCK-8), flow cytometry assays, ELISA and reactive oxygen species (ROS) and malondialdehyde (MDA) detection, were performed to assess cell viability, apoptosis, inflammation and oxidative stress, respectively. Western blot analysis was carried out to examine the protein levels of Bax, Bcl-2, and SNHG6. Luciferase reporter and RIP assays were used to confirm the true interaction between SNHG6 and miR-135a-5p, or miR-135a-5p and ROCK.
The levels of SNHG6, ROCK1 and ROCK2 were notably increased and miR-135a-5p was decreased in atherosclerosis patients and oxidized low-density lipoprotein (ox-LDL)-treated HUVECs. Knockdown of SNHG6 alleviated ox-LDL-induced injury of HUVECs, while this effect was partly reversed by miR-135a-5p inhibitor. Moreover, overexpression of ROCKs aggravated miR-135a-5p-alleviated atherosclerosis cell injury. SNHG6 contributed to ROCK expression through sequestering miR-135a-5p as a molecular sponge.
SNHG6 functions as a promoter in atherosclerosis events by targeting miR-135a-5p/ROCK axis in ox-LDL-stimulated HUVECs. This finding will help to develop a novel therapeutic strategy for atherosclerosis.
大量长链非编码RNA（lncRNA）在动脉粥样硬化进展中发挥重要作用。小核仁RNA宿主基因6（SNHG6）是一种著名的lncRNA，在动脉粥样硬化患者中异常高表达。然而，其在动脉粥样硬化事件中的功能和基本机制尚未完全阐明。
通过RT-qPCR检测临床样本和细胞中SNHG6、miR-135a-5p、ROCK1和ROCK2的表达模式。分别进行细胞计数试剂盒-8（CCK-8）、流式细胞术检测、ELISA以及活性氧（ROS）和丙二醛（MDA）检测，以评估细胞活力、凋亡、炎症和氧化应激。进行蛋白质印迹分析以检测Bax、Bcl-2和SNHG6的蛋白质水平。使用荧光素酶报告基因和RNA免疫沉淀（RIP）实验来确认SNHG6与miR-135a-5p之间，或miR-135a-5p与ROCK之间的真实相互作用。
在动脉粥样硬化患者和氧化型低密度脂蛋白（ox-LDL）处理的人脐静脉内皮细胞（HUVECs）中，SNHG6、ROCK1和ROCK2水平显著升高，而miR-135a-5p水平降低。敲低SNHG6可减轻ox-LDL诱导的HUVECs损伤，而miR-135a-5p抑制剂可部分逆转这种作用。此外，ROCKs的过表达加剧了miR-135a-5p减轻的动脉粥样硬化细胞损伤。SNHG6作为分子海绵通过隔离miR-135a-5p促进ROCK表达。
在ox-LDL刺激的HUVECs中，SNHG6通过靶向miR-135a-5p/ROCK轴在动脉粥样硬化事件中起促进作用。这一发现将有助于开发一种新的动脉粥样硬化治疗策略。
