An Rui, Xi Cong, Xu Jian, Liu Ying, Zhang Shumiao, Wang Yuemin, Hao Yuewen, Sun Lijun
Department of Radiology, Xijing Hospital, Fourth Military Medical University, Xi'an, China.
Department of Neurology, Baoji City People's Hospital, Baoji, China.
Oxid Med Cell Longev. 2017;2017:1271670. doi: 10.1155/2017/1271670. Epub 2017 Mar 2.
Cotransfer of angiogenic and antiapoptotic genes could be the basis of new gene therapy strategies for myocardial infarction. In this study, rAAV-PR39-ADM, coexpressing antimicrobial peptide (PR39) and adrenomedullin (ADM), was designed with the mediation of recombinant adeno-associated virus. In vitro, CRL-1730 cells were divided into four groups, namely, the sham group, the AAV-null group, the NS (normal saline) group, and the PR39-ADM group. Immunocytochemistry analysis, CCK-8 assays, Matrigel assays, and apoptotic analysis were performed; in vivo, myocardial infarction model was established through ligation of the left coronary artery on rats, and treatment groups corresponded to those used in vitro. Myocardial injury, cardiac performance, and the extent of myocardial apoptosis were assessed. Results suggested that rAAV-PR39-ADM administration after myocardial infarction improved cell viability and cardiac function, attenuated apoptosis and myocardial injury, and promoted angiogenesis. Subsequently, levels of 6×His, HIF-1, VEGF, p-Akt, Akt, ADM, Bcl-2, and Bax were measured by western blot. rAAV-PR39-ADM increased p-Akt, HIF-1, and VEGF levels and induced higher Bcl-2 expression and lower Bax expression. In conclusion, our results demonstrate that rAAV-PR39-ADM mitigates myocardial injury by promoting angiogenesis and reducing apoptosis. This study suggests a potential novel gene therapy-based method that could be used clinically for myocardial infarction.
血管生成基因与抗凋亡基因的共转移可能成为心肌梗死新基因治疗策略的基础。在本研究中，在重组腺相关病毒的介导下，设计了共表达抗菌肽（PR39）和肾上腺髓质素（ADM）的重组腺相关病毒载体rAAV-PR39-ADM。体外实验中，将CRL-1730细胞分为四组，即假手术组、空腺相关病毒组、生理盐水组和PR39-ADM组。进行了免疫细胞化学分析、CCK-8检测、基质胶实验和凋亡分析；体内实验中，通过结扎大鼠左冠状动脉建立心肌梗死模型，治疗组与体外实验相同。评估心肌损伤、心脏功能和心肌凋亡程度。结果表明，心肌梗死后给予rAAV-PR39-ADM可提高细胞活力和心脏功能，减轻凋亡和心肌损伤，并促进血管生成。随后，通过蛋白质免疫印迹法检测6×His、HIF-1、VEGF、p-Akt、Akt、ADM、Bcl-2和Bax的水平。rAAV-PR39-ADM可提高p-Akt、HIF-1和VEGF水平，并诱导更高的Bcl-2表达和更低的Bax表达。总之，我们的结果表明rAAV-PR39-ADM通过促进血管生成和减少凋亡来减轻心肌损伤。本研究提示了一种潜在的基于新基因治疗的方法，可用于心肌梗死的临床治疗。
