It is reported that circular RNAs (circRNAs) play a key role in atherosclerosis (AS). Foam cell formation, which is the main feature of AS, can be significantly inhibited by cholesterol efflux. We established a model of astaxanthin (AST) promoting cholesterol efflux from macrophages through oil red O staining, real-time quantitative PCR (qRT-PCR), and western blot and used RNA sequencing to detect the expression of circRNAs in AST-treated and untreated THP-1 cells. Finally, siRNA transfection screened out circRNAs that were significantly differentially expressed. The data analysis was performed by Student’s t test and P < 0.05 was considered statistically significant. In the model of AST promoting cholesterol efflux from THP-1 cells, there were a total of 7276 circRNAs differentially expressed, among which the top 25 upregulated and the top 25 downregulated circRNAs were selected based on the log2 (fold change). GO analysis showed that differential expression of circRNAs in biological process (2066/3098; 66.69%), molecular function (543/3098; 17.53%), and cellular component (489/3098; 15.78%). Based on KEGG analysis, RNA transport was the most enriched pathway. Finally, we obtained 3 significantly upregulated circRNAs by siRNA transfection and qRT-PCR. The 3 differentially expressed circRNAs may play an important role in the process of AST promoting cholesterol efflux and may be used as biomarkers to prevent AS.

中文翻译：

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基于RNA-seq的虾青素中环状RNA的表达促进胆固醇从THP-1细胞流出

据报道，环状RNA（circRNA）在动脉粥样硬化（AS）中起关键作用。泡沫细胞的形成是 AS 的主要特征，可被胆固醇外流显着抑制。我们通过油红 O 染色、实时定量 PCR (qRT-PCR) 和蛋白质印迹建立了虾青素 (AST) 促进巨噬细胞胆固醇流出的模型，并使用 RNA 测序检测 AST 处理和未处理中 circRNA 的表达THP-1 细胞。最后，siRNA转染筛选出显着差异表达的circRNA。数据分析通过Student's t 检验进行，P < 0.05 被认为具有统计学意义。在AST促进THP-1细胞胆固醇外流模型中，共有7276个circRNA差异表达，其中根据log2（倍数变化）选择前25个上调和前25个下调的circRNA。GO分析显示circRNA在生物过程（2066/3098；66.69%）、分子功能（543/3098；17.53%）和细胞成分（489/3098；15.78%）中的差异表达。基于 KEGG 分析，RNA 转运是最丰富的途径。最后，我们通过siRNA转染和qRT-PCR获得了3个显着上调的circRNA。这3个差异表达的circRNA可能在AST促进胆固醇外流的过程中发挥重要作用，可作为预防AS的生物标志物。和细胞成分（489/3098；15.78%）。基于 KEGG 分析，RNA 转运是最丰富的途径。最后，我们通过siRNA转染和qRT-PCR获得了3个显着上调的circRNA。这3个差异表达的circRNA可能在AST促进胆固醇外流的过程中发挥重要作用，可作为预防AS的生物标志物。和细胞成分（489/3098；15.78%）。基于 KEGG 分析，RNA 转运是最丰富的途径。最后，我们通过siRNA转染和qRT-PCR获得了3个显着上调的circRNA。这3个差异表达的circRNA可能在AST促进胆固醇外流的过程中发挥重要作用，可作为预防AS的生物标志物。