saRNA guided iNOS up-regulation improves erectile function of diabetic rats.
saRNA guided iNOS up-regulation improves erectile function of diabetic rats.
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DOI：
10.1016/j.juro.2013.03.043
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发表时间：
2013-08
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影响因子：
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通讯作者：
Tao Wang;Ming-chao Li;Huixing Yuan;Y. Zhan;Hua Xu;Shaogang Wang;Wei-min Yang;Jihong Liu;Z. Ye;Long-cheng Li
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文献类型：
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作者：
Tao Wang;Ming-chao Li;Huixing Yuan;Y. Zhan;Hua Xu;Shaogang Wang;Wei-min Yang;Jihong Liu;Z. Ye;Long-cheng Li
目的启动子靶向 saRNA 介导基因表达的序列特异性上调。我们探索了 RNA 激活介导的 iNOS 基因激活对改善糖尿病大鼠模型勃起功能的治疗作用。材料和方法将 iNOS 启动子特异的最佳 saRNA 序列克隆到腺病毒载体中，产生 AdU6/shiNOS 和 AdU6/shControl。使用相应的病毒转导培养的大鼠海绵体平滑肌细胞。在大鼠中建立链脲佐菌素诱导的糖尿病模型，并用于测试海绵体内递送 iNOS saRNA 病毒对勃起功能的影响。通过逆转录聚合酶链反应和蛋白质印迹评估治疗大鼠海绵体平滑肌细胞或阴茎组织中 iNOS 的表达。通过酶联免疫吸附测定法分析环状单磷酸鸟苷。在注射后第 1、3、5、7、10 和 14 天，使用数据采集系统测量对海绵体神经刺激的海绵体内压力。结果腺病毒介导的 iNOS saRNA 表达引起海绵体平滑肌细胞中 iNOS 的持续上调。海绵体内注射AdU6/shiNOS通过一氧化氮/细胞内环单磷酸鸟苷激活激活链脲佐菌素诱导的糖尿病大鼠体内iNOS表达，并显着增加海绵体内峰值压力。结论结果表明saRNA介导的阴茎iNOS过表达可以通过一氧化氮-环单磷酸鸟苷途径恢复链脲佐菌素糖尿病大鼠的勃起功能。
PurposePromoter targeted saRNAs mediate sequence specific up-regulation of gene expression. We explored the therapeutic effect of RNA activation mediated iNOS gene activation on improving erectile function in a rat model of diabetes mellitus.Materials and MethodsAn optimal saRNA sequence specific for iNOS promoter was cloned into an adenoviral vector, resulting in AdU6/shiNOS and AdU6/shControl. The corresponding viruses were used to transduce cultured rat cavernous smooth muscle cells. Streptozotocin induced diabetes models were established in rats and used to test the effects of intracavernous delivery of iNOS saRNA viruses on erectile function. iNOS expression in the cavernous smooth muscle cells or penile tissue of treated rats was assessed by reverse transcriptase-polymerase chain reaction and Western blot. Cyclic guanosine monophosphate was analyzed by enzyme-linked immunosorbent assay. Intracavernous pressure in response to cavernous nerve stimulation was measured using a data acquisition system on post-injection days 1, 3, 5, 7, 10 and 14.ResultsAdenovirus mediated expression of iNOS saRNA caused sustained up-regulation of iNOS in cavernous smooth muscle cells. Intracavernous injection of AdU6/shiNOS activated iNOS expression in vivo and significantly increased peak intracavernous pressure in streptozotocin induced diabetic rats via nitric oxide/intracellular cyclic guanosine monophosphate activation.ConclusionsResults show that saRNA mediated iNOS over expression in the penis can restore erectile function in streptozocin diabetic rats via the nitric oxide-cyclic guanosine monophosphate pathway.