Department of Cardiothoracic Surgery, Changhai Hospital, Second Military Medical University, Shanghai, People's Republic of China.
Department of Cardiothoracic Surgery, Changhai Hospital, Second Military Medical University, Shanghai, People's Republic of China.
J Thorac Cardiovasc Surg. 2015 Nov;150(5):1292-301.e2. doi: 10.1016/j.jtcvs.2015.08.010. Epub 2015 Aug 8.
To elucidate the mechanisms of Brahma-related gene 1 (Brg1) involvement in the pathophysiologic processes of aortic dissection.
Seventeen dissecting, 4 dilated, and 10 healthy human aorta samples were collected. Expression of Brg1 in the medium of aorta was evaluated by quantitative real-time polymerase chain reaction, Western blot, and immunohistochemical staining, respectively. The regulation effect of Brg1 on proliferation and migration of human aortic smooth muscle cells (HASMCs) was analyzed in 3 ways: using cell counting, a migration chamber, and a wound scratch assay. A polymerase chain reaction array was used for screening potential target genes of Brg1. A chromatin immunoprecipitation assay was adopted for direct deoxyribonucleic acid-protein binding detection.
Expression levels of Brg1 were increased in aortic dissection and aortic dilation patients. In vitro results indicated that overexpression of Brg1 inhibited proliferation and migration of HASMCs. The candidate proliferation- and migration-related Brg1 target gene found was Ras-related associated with diabetes (RRAD), expression levels of which were enhanced in dissecting aortic specimens. The direct regulation effect of Brg1 on RRAD was verified by chromatin immunoprecipitation assay results. Furthermore, down-regulating RRAD significantly alleviated the suppression effects of Brg1 on proliferation and migration of HASMCs.
Our study illustrated that Brg1 inhibited the proliferation and migration capacity of HASMCs, via the mechanism of direct up-regulation of RRAD, thus playing an important role in the pathophysiologic processes of aortic dissection.
阐明 Brg1 参与主动脉夹层病理生理过程的机制。
收集 17 例夹层、4 例扩张和 10 例健康人主动脉标本。分别采用实时定量聚合酶链反应、Western blot 和免疫组织化学染色法评估 Brg1 在主动脉中的表达。采用细胞计数、迁移室和划痕实验 3 种方法分析 Brg1 对人主动脉平滑肌细胞(HASMC)增殖和迁移的调节作用。采用聚合酶链反应阵列筛选 Brg1 的潜在靶基因。采用染色质免疫沉淀法进行直接 DNA-蛋白质结合检测。
Brg1 的表达水平在主动脉夹层和主动脉扩张患者中增加。体外结果表明,Brg1 过表达抑制 HASMC 的增殖和迁移。发现的候选增殖和迁移相关 Brg1 靶基因 Ras-related associated with diabetes (RRAD)在夹层主动脉标本中的表达增强。染色质免疫沉淀实验结果验证了 Brg1 对 RRAD 的直接调节作用。此外,下调 RRAD 显著减轻了 Brg1 对 HASMC 增殖和迁移的抑制作用。
本研究表明,Brg1 通过直接上调 RRAD 抑制 HASMC 的增殖和迁移能力,从而在主动脉夹层的病理生理过程中发挥重要作用。