Up-regulation of VEGF by Small Activator RNA in Human Corpus Cavernosum Smooth Muscle Cells
Up-regulation of VEGF by Small Activator RNA in Human Corpus Cavernosum Smooth Muscle Cells
复制标题
小激活子 RNA 对人海绵体平滑肌细胞中 VEGF 的上调
DOI：
10.1111/j.1743-6109.2011.02412.x
复制
发表时间：
2011-10-01
影响因子：
3.5
通讯作者：
Ye, Zhangqun
中科院分区：
文献类型：
--
作者：
Chen, Ruibao;Wang, Tao;Ye, Zhangqun
导论.阴茎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（艾德）。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，因此预期增加VEGF的表达可能对勃起功能具有有益作用。本文旨在探讨核糖核酸激活（RNAa）技术诱导VEGF表达的可能性，以及RNAa诱导VEGF表达治疗ED的可能性。原代分离培养人阴茎海绵体平滑肌细胞（CCSMCs）。免疫组化法检测α-平滑肌肌动蛋白的表达，鉴定CCSMC。化学合成先前鉴定的VEGF启动子靶向的小激活物RNA（saRNA，双链[ds] VEGF-706）和阴性对照dsRNA。用saRNA转染培养的人CCSMC。RT-PCR和Western blotting检测转染后CCSMCs中VEGF mRNA和蛋白的表达。免疫荧光染色也被用来确认VEGF蛋白在培养的CCSMCs中的表达。采用RT定量PCR、免疫印迹和免疫荧光法检测VEGF的表达。转染dsVEGF-706后，RT定量PCR分析显示转染dsVEGF-706的细胞VEGF mRNA的表达明显高于对照组（P < 0.05）。Western blotting和免疫荧光分析结果表明，dsVEGF-706也能诱导VEGF蛋白的表达，与mRNA的诱导一致。VEGF的表达可以在原代人CCSMC中被RNA激活，这表明RNA介导的VEGF激活在治疗ED中的潜在应用。Chen R，Wang T，Rao K，Yang J，Zhang S，Wang S，Liu J，and Ye Z.小分子激活RNA上调人阴茎海绵体平滑肌细胞VEGF表达J Sex Med 2011; 8：2773-2780.
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. Chen R, Wang T, Rao K, Yang J, Zhang S, Wang S, Liu J, and Ye Z. Up-regulation of VEGF by small activator RNA in human corpus cavernosum smooth muscle cells. J Sex Med 2011; 8: 2773-2780.