Zhang S C, Chen L L, Wang W J, Wu X G, Zhou X
Department of Cardiology, Second Affiliated Hospital of Soochow University, Suzhou 215004, China.
Experimental Center, Second Affiliated Hospital of Soochow University, Suzhou 215004, China.
Zhonghua Xin Xue Guan Bing Za Zhi. 2021 Nov 24;49(11):1130-1138. doi: 10.3760/cma.j.cn112148-20210929-00838.
To explore the differential expression of circRNAs and their potential impact on the pathophysiological process in cardiac hypertrophy. Six SPF C57BL/6J male mice, aged 8 to 10 weeks, were randomly divided into transverse aortic constriction (TAC) group (=3) or sham operation(sham) group (=3) according to random number table method. TAC mouse model was used to induce cardiac hypertrophy. Four weeks after surgery, high-throughput sequencing analysis was performed to detect differentially expressed circRNA in left myocardial tissues of mice between TAC group and sham group, and principal component analysis of circRNA was performed by R language software. Enrichment analysis was performed by GO and KEGG databases to predict the basic functions of differentially expressed circRNA-derived genes and their biological pathways. The differentially expressed circRNAs in the sequencing results were verified by real-time fluorescence quantitative polymerase chain reaction. Cytoscape software was used to construct circRNA-microRNA (miRNA) network maps to predict their interactions by combining differentially expressed circRNA and TargetScan predicted miRNA sites. Principal component analysis was performed on 4 580 circRNAs detected from 6 samples of mice in TAC group and sham group. The results of R language software indicated that the variance contribution rate of the first 3 principal components, namely the first, second and third principal components, was 91.01%, 3.19% and 2.01%, respectively, and the cumulative variance contribution rate of the 3 components was 96.21%. Among the differentially expressed circRNAs, 6 (19%) were up-regulated and 25 (81%) were down-regulated in the TAC group. GO analysis showed that differentially expressed circRNA was closely related to the occurrence and development of cardiac hypertrophy, and KEGG pathway analysis suggested that downregulated circRNA expression was involved in the regulation of actin cytoskeleton. Fifteen out of the 31 differentially expressed circRNAs were selected for real-time fluorescence quantitative polymerase chain reaction verification, and the results showed that 8 circRNAs were consistent with sequencing results. circRNA-miRNA co-expression network analysis results showed that chr11:65218529-65233184-interacts with mmu-miRNA-30e-3p and mmu-miRNA-30a-3p. s The differential expression of circRNA in hypertrophic myocardium mice is evidenced in TAC mouse model. circRNA may interact with the corresponding miRNA to influence the occurrence and development of cardiac hypertrophy through autophagy-related cellular hypertrophy pathway or apoptosis-related pathological phenotypes.
为探讨环状RNA（circRNA）的差异表达及其对心肌肥厚病理生理过程的潜在影响。选取6只8至10周龄的SPF级C57BL/6J雄性小鼠，根据随机数字表法随机分为横断主动脉缩窄（TAC）组（n = 3）或假手术（sham）组（n = 3）。采用TAC小鼠模型诱导心肌肥厚。术后4周，进行高通量测序分析，以检测TAC组和假手术组小鼠左心室心肌组织中差异表达的circRNA，并使用R语言软件对circRNA进行主成分分析。通过GO和KEGG数据库进行富集分析，以预测差异表达的circRNA衍生基因的基本功能及其生物学途径。通过实时荧光定量聚合酶链反应验证测序结果中差异表达的circRNA。使用Cytoscape软件构建circRNA-微小RNA（miRNA）网络图，通过结合差异表达的circRNA和TargetScan预测的miRNA位点来预测它们之间的相互作用。对TAC组和假手术组6只小鼠样本中检测到的4580个circRNA进行主成分分析。R语言软件结果显示，前3个主成分，即第一、第二和第三主成分的方差贡献率分别为91.01%、3.19%和2.01%，3个成分的累积方差贡献率为96.21%。在差异表达的circRNA中，TAC组有6个（19%）上调，25个（81%）下调。GO分析表明，差异表达的circRNA与心肌肥厚的发生发展密切相关，KEGG通路分析提示下调的circRNA表达参与肌动蛋白细胞骨架的调节。从31个差异表达的circRNA中选取15个进行实时荧光定量聚合酶链反应验证，结果显示8个circRNA与测序结果一致。circRNA-miRNA共表达网络分析结果显示，chr11:65218529-65233184与mmu-miRNA-30e-3p和mmu-miRNA-30a-3p相互作用。在TAC小鼠模型中证实了肥厚心肌小鼠中circRNA的差异表达。circRNA可能与相应的miRNA相互作用，通过自噬相关的细胞肥大途径或凋亡相关的病理表型影响心肌肥厚的发生发展。
