腺相关病毒载体递送和低氧反应元件调节缺血性大鼠心脏中 CD151 的表达。
Adeno associated viral vector-delivered and hypoxia response element-regulated CD151 expression in ischemic rat heart.
机构信息
Department of Rehabilitation Medicine of Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
出版信息
Acta Pharmacol Sin. 2011 Feb;32(2):201-8. doi: 10.1038/aps.2010.205. Epub 2011 Jan 17.
AIM
The aim of this study was to improve the delivery efficacy and target specificity of the pro-angiogenic gene CD151 to the ischemic heart.
METHODS
To achieve the inducible expression of adeno-associated viral (AAV)-delivered CD151 gene in only the ischemic myocardium, we generated an AAV construct in which CD151 expression can be controlled by the hypoxia response element (HRE) sequence from the human Enolase gene. The function of this vector was examined in rat H9C2 cardiac myoblasts and in ischemic rat myocardium. The expression of CD151 in the areas of ischemic myocardium was confirmed at the mRNA level by real-time PCR and on the protein level by Western blot, whereas the CD151 expression in the microvessels within the areas of ischemic myocardium was detected by immunohistochemistry.
RESULTS
HRE significantly enhances the expression of CD151 under hypoxic conditions or in the ischemic myocardium, and forced CD151 expression increases the number of microvessels in the ischemic myocardium.
CONCLUSION
The AAV-mediated, HRE regulated delivery of the CD151 gene shows higher expression in the ischemic myocardium and more efficiently targets CD151 to the hypoxic regions after myocardial infarction.
目的
本研究旨在提高促血管生成基因 CD151 向缺血性心脏的传递效果和靶向特异性。
方法
为了实现在缺血心肌中仅诱导表达腺相关病毒（AAV）递送的 CD151 基因，我们构建了一种 AAV 载体，其中 CD151 的表达可以受人类烯醇化酶基因的缺氧反应元件（HRE）序列的控制。该载体的功能在大鼠 H9C2 心肌细胞和缺血性大鼠心肌中进行了检测。通过实时 PCR 在 mRNA 水平上，通过 Western blot 在蛋白水平上，以及通过免疫组织化学检测缺血心肌区域内微血管中的 CD151 表达，证实了 CD151 在缺血心肌区域的表达。
结果
HRE 在缺氧条件下或在缺血心肌中显著增强 CD151 的表达，强制表达 CD151 增加了缺血心肌中的微血管数量。
结论
AAV 介导的、HRE 调节的 CD151 基因传递在缺血心肌中的表达更高，并且在心肌梗死后更有效地将 CD151 靶向到缺氧区域。
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