Intramyocardial Injection of Recombinant Adeno-Associated Viral Vector Coexpressing PR39/Adrenomedullin Enhances Angiogenesis and Reduces Apoptosis in a Rat Myocardial Infarction Model.
Intramyocardial Injection of Recombinant Adeno-Associated Viral Vector Coexpressing PR39/Adrenomedullin Enhances Angiogenesis and Reduces Apoptosis in a Rat Myocardial Infarction Model.
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心肌内注射共表达 PR39/肾上腺髓质素的重组腺相关病毒载体可增强大鼠心肌梗死模型中的血管生成并减少细胞凋亡
DOI：
10.1155/2017/1271670
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发表时间：
2017
影响因子：
--
通讯作者：
Sun L
中科院分区：
文献类型：
--
作者：
An R;Xi C;Xu J;Liu Y;Zhang S;Wang Y;Hao Y;Sun L
血管生成和抗凋亡基因的共转移可能成为心肌梗死基因治疗新策略的基础。本研究利用重组腺相关病毒介导，设计了共表达抗菌肽（PR 39）和肾上腺髓质素（ADM）的rAAV-PR 39-ADM。体外实验将CRL-1730细胞分为4组，即假手术组（sham）、AAV空载体组（AAV null）、生理盐水组（NS）和PR 39-ADM组。进行免疫细胞化学分析、CCK-8测定、Matrigel测定和细胞凋亡分析;在体内，通过结扎大鼠左冠状动脉建立心肌梗死模型，并且处理组对应于体外所用的那些。评估心肌损伤、心脏功能和心肌细胞凋亡程度。结果表明，心肌梗死后给予rAAV-PR 39-ADM可改善细胞活力和心功能，减轻细胞凋亡和心肌损伤，并促进血管生成。Western blot检测6×His、HIF-1α、VEGF、p-Akt、Akt、ADM、Bcl-2和Bax的表达。rAAV-PR 39-ADM可增加p-Akt、HIF-1α和VEGF的表达，并诱导Bcl-2的高表达和Bax的低表达。总之，我们的结果表明，rAAV-PR 39-ADM通过促进血管生成和减少细胞凋亡减轻心肌损伤。这项研究提出了一种潜在的新的基因治疗为基础的方法，可用于临床心肌梗死。
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-1α, VEGF, p-Akt, Akt, ADM, Bcl-2, and Bax were measured by western blot. rAAV-PR39-ADM increased p-Akt, HIF-1α, 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.