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首页> 外文期刊>The journal of sexual medicine >Up-regulation of VEGF by small activator RNA in human corpus cavernosum smooth muscle cells.
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Up-regulation of VEGF by small activator RNA in human corpus cavernosum smooth muscle cells.

机译：在人海绵体平滑肌细胞中，小激活RNA上调VEGF。

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INTRODUCTION: Functional failure of smooth muscle cells and endothelial cells in corpus cavernosum contributes to erectile dysfunction (ED) in aging men. Given that vascular endothelial growth factor (VEGF) may improve the function of smooth muscle cells and endothelial cells through different mechanisms, it is thus expected that increasing the expression of VEGF may have beneficial effects on erectile function. AIM: The aim of this article is to explore the possibility that VEGF can be induced by ribonucleic acid activation (RNAa) technology, and VEGF induction by RNAa has the potential of treating ED. METHODS: Primary human corpus cavernosum smooth muscle cells (CCSMCs) were isolated and cultured in vitro. The expression of alpha-smooth muscle actin was detected by immunohistochemistry to identify CCSMCs. A previously identified VEGF promoter-targeted small activator RNA (saRNA, double-stranded [ds]VEGF-706) and a negative control dsRNA were chemically synthesized. Cultured human CCSMCs were transfected with the saRNAs. The expression of VEGF messenger RNA (mRNA) and protein in transfected CCSMCs was evaluated by real-time polymerase chain reaction (RT-PCR) and Western blotting assay, respectively. Immunofluorescent staining was also used to confirm VEGF protein expression in cultured CCSMCs. MAIN OUTCOME MEASURE: The expression of VEGF was assessed by RT quantitative PCR, Western blotting, and immunofluorescence assays. RESULTS: After transfection, RT quantitative PCR analysis showed that the expression of VEGF mRNA was significantly induced in dsVEGF-706 transfected cells compared with cells receiving control treatments (P < 0.05). Consistent with mRNA induction, Western blotting and immunofluorescence analysis showed that VEGF protein expression was also induced by dsVEGF-706. CONCLUSION: VEGF expression can be activated by RNAa in primary human CCSMCs, suggesting a potential application of RNAa-mediated VEGF activation for the treatment of ED.
机译：引言：海绵体中平滑肌细胞和内皮细胞的功能衰竭导致老年男性勃起功能障碍（ED）。鉴于血管内皮生长因子（VEGF）可以通过不同的机制改善平滑肌细胞和内皮细胞的功能，因此可以预期增加VEGF的表达可能对勃起功能产生有益作用。目的：本文的目的是探讨核糖核酸激活（RNAa）技术可诱导VEGF的可能性，而RNAa诱导VEGF具有治疗ED的潜力。方法：分离并培养原代人海绵体平滑肌细胞（CCSMC）。通过免疫组织化学检测α平滑肌肌动蛋白的表达以鉴定CCSMC。化学合成先前鉴定的靶向VEGF启动子的小激活RNA（saRNA，双链[ds] VEGF-706）和阴性对照dsRNA。用saRNA转染培养的人CCSMC。分别通过实时聚合酶链反应（RT-PCR）和蛋白质印迹法评估转染CCSMCs中VEGF Messenger mRNA（mRNA）和蛋白的表达。免疫荧光染色也用于确认培养的CCSMC中VEGF蛋白的表达。主要观察指标：通过RT定量PCR，Western印迹和免疫荧光分析评估VEGF的表达。结果：转染后，RT定量PCR分析显示，与接受对照的细胞相比，dsVEGF-706转染的细胞明显诱导了VEGF mRNA的表达（P <0.05）。与mRNA诱导一致，蛋白质印迹和免疫荧光分析表明dsVEGF-706也诱导VEGF蛋白表达。结论：RNAa可在人原代CCSMCs中激活VEGF的表达，提示RNAa介导的VEGF激活在ED治疗中的潜在应用。

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