Jin Xin, Pan Zhengxia, Zhao Zhenjiang, Ouyang Da, Qin Jinjie, Tian Jie
National Clinical Research Center for Child Health and Disorders, Ministry of Education Key Laboratory of Child Development, Chongqing Key Laboratory of Structural Birth Defect and Reconstruction, Children's Hospital of Chongqing Medical University, Chongqing, PR China.
Department of Cardiac Surgery Children's Hospital of Chongqing Medical University, Chongqing, PR China.
Heliyon. 2024 Nov 20;10(23):e40512. doi: 10.1016/j.heliyon.2024.e40512. eCollection 2024 Dec 15.
(1) ObjectiveCoarctation of the aorta (CoA) is a complex congenital heart disease. Research on differential genes in patients with CoA and other groups of aortas and investigating the pathogenesis of aorta coarctation is essential for prevention and diagnosis. This study was conducted between January 2019 and December 2021. The first step was the analysis of differential genes in four groups of aortic tissues. Group A: 46 cases, coarctation specimen of the CoA group; Group B: 46 cases, anastomotic margin specimen of the CoA group; Group C: 22 cases, aortic specimen of the aortic stenosis group; and Group D: 6 cases, aorta of the necropsy group. Genomic and proteomic differences were compared in aortic specimens from different patient groups in Southwestern China. The second step the CRISPR technology was used to knock out the differential genes in mice. The phenotypes were verified using ultrasound after obtaining homozygous mice. : Quantitative polymerase chain reaction indicated that BMP4, Smad4, STRA6, CRABP I, and COX-2 genes were statistically downregulated in group A than in other groups. STRA6 expression was 0.33 ± 0.66 in group A and 1.03 ± 0.20 in group D. The difference between the two groups was statistically significant. Western blotting revealed that BMP4, Smad4, STRA6, and CRABP I had a low expression in group A at the proteomic level. STRA6 protein was 0.13 ± 0.02 in group A and 0.48 ± 0.07 in group D. Group A decreased by 73.7 %, a statistically significant difference. Double-labeled immunofluorescence demonstrated co-expression of BMP4 and STRA6 in the tunica adventitia vasorum of group A. However, BMP4 expression in the tunica adventitia vasorum and endangium of the constricted aorta showed a double-loop sign. Primary filial generation offspring were generated using CRISPR knockout of the STRA6 gene. The first filial generation was mated with the wild-type mice to produce the second filial generation. 72.7 % homozygous mice in the fourth week of the second filial generation were detected using ultrasound, indicating a reduction in aortic diameter. (4) : BMP4, STRA6, and COX-2 genes may be associated with CoA in Southwestern Chinese patients. STRA6 knockout, a gene that initiates the retinoic acid pathway, may lead to CoA.
（1）目的：主动脉缩窄（CoA）是一种复杂的先天性心脏病。研究CoA患者与其他主动脉组的差异基因并探讨主动脉缩窄的发病机制对预防和诊断至关重要。本研究于2019年1月至2021年12月进行。第一步是分析四组主动脉组织中的差异基因。A组：46例，CoA组缩窄标本；B组：46例，CoA组吻合缘标本；C组：22例，主动脉狭窄组主动脉标本；D组：6例，尸检组主动脉。比较中国西南部不同患者组主动脉标本中的基因组和蛋白质组差异。第二步，使用CRISPR技术敲除小鼠中的差异基因。获得纯合小鼠后，用超声验证表型。定量聚合酶链反应表明，A组中BMP4、Smad4、STRA6、CRABP I和COX-2基因在统计学上比其他组下调。A组中STRA6表达为0.33±0.66，D组为1.03±0.20。两组间差异有统计学意义。蛋白质印迹法显示，A组中BMP4、Smad4、STRA6和CRABP I在蛋白质组水平表达较低。A组中STRA6蛋白为0.13±0.02，D组为0.48±0.07。A组下降了73.7%，差异有统计学意义。双标免疫荧光显示A组血管外膜中BMP4和STRA6共表达。然而，在缩窄主动脉的血管外膜和内膜中BMP4表达呈双环征。使用CRISPR敲除STRA6基因产生初代子代后代。第一代与野生型小鼠交配产生第二代。第二代第四周时，用超声检测到72.7%的纯合小鼠，表明主动脉直径减小。（4）结论：BMP4、STRA6和COX-2基因可能与中国西南部患者的CoA有关。启动视黄酸途径的基因STRA6敲除可能导致CoA。
