Purpose

Atherosclerosis is the leading cause of cardiovascular diseases (CVD) with high incidence rate and mortality rate. Long non-coding RNAs (lncRNAs) are important functional molecules in atherosclerosis. Present study aimed to explore the functional role and underlying mechanism of ZFAS1 in atherosclerosis.

Methods

The in-vitro cell model of atherosclerosis was established by using oxidized low-density lipoprotein (ox-LDL) to induce THP-1 macrophage-derived foam cells. qRT-PCR measured the mRNA levels of ZFAS1, miR-654-3p, ADAM10 and RAB22A. Western blot detected the protein levels of ADAM10 and RAB22A. The levels of IL-1β, IL-6 and TNF-ɑ (inflammatory biomarkers) were tested with ELISA assay. Detection of cholesterol efflux rate was experimented. The interaction between RNAs was affirmed with luciferase reporter and RNA pull-down experiments.

Results

The expression of ZFAS1 was significantly up-regulated in in-vitro cell model of atherosclerosis at a dose- and time-dependent manner. Knockdown of ZFAS1 impaired inflammatory responses and promoted cholesterol efflux rate. Overexpression of ZFAS1 accelerated inflammatory responses and hampered cholesterol efflux rate. Then, the cytoplasmic role of ZFAS1 was revealed. By bio-informatics analysis and mechanism assays, miR-654-3p was identified to bind with ZFAS1. Moreover, ADAM10 and RAB22A were targeted and suppressed by miR-654-3p. ZFAS1 served as a ceRNA to positively regulate ADAM10 and RAB22A expression through endogenously sponging miR-654-3p.

Conclusion

In conclusion, ZFAS1 elevated ADAM10/RAB22A expression to reduce cholesterol efflux rate and facilitate inflammatory responses in atherosclerosis at a miR-654-3p-dependent way, suggesting a prospective treatment method for amelioration of atherosclerosis.

中文翻译：

---

LncRNA ZFAS1 通过调节 miR-654-3p-ADAM10/RAB22A 轴赋予炎症反应并减少动脉粥样硬化中的胆固醇流出。

目的

动脉粥样硬化是心血管疾病（CVD）的主要原因，发病率和死亡率都很高。长链非编码 RNA (lncRNA) 是动脉粥样硬化中重要的功能分子。本研究旨在探讨 ZFAS1 在动脉粥样硬化中的功能作用和潜在机制。

方法

利用氧化低密度脂蛋白（ox-LDL）诱导THP-1巨噬细胞来源的泡沫细胞建立了动脉粥样硬化的体外细胞模型。qRT-PCR 测量了 ZFAS1、miR-654-3p、ADAM10 和 RAB22A 的 mRNA 水平。Western blot检测ADAM10和RAB22A的蛋白水平。用 ELISA 法检测 IL-1β、IL-6 和 TNF-ɑ（炎症生物标志物）的水平。对胆固醇流出率的检测进行了实验。荧光素酶报告基因和 RNA 下拉实验证实了 RNA 之间的相互作用。

结果

ZFAS1的表达在动脉粥样硬化体外细胞模型中以剂量和时间依赖性方式显着上调。ZFAS1 的敲低会损害炎症反应并促进胆固醇流出率。ZFAS1 的过表达加速炎症反应并阻碍胆固醇流出率。然后，揭示了 ZFAS1 的细胞质作用。通过生物信息学分析和机制分析，鉴定出miR-654-3p与ZFAS1结合。此外，ADAM10 和 RAB22A 被 miR-654-3p 靶向和抑制。ZFAS1 作为 ceRNA 通过内源性海绵化 miR-654-3p 正向调节 ADAM10 和 RAB22A 的表达。

结论

总之，ZFAS1 以 miR-654-3p 依赖的方式提高 ADAM10/RAB22A 的表达以降低胆固醇流出率并促进动脉粥样硬化中的炎症反应，提示一种改善动脉粥样硬化的前瞻性治疗方法。