title: saRNA guided iNOS up-regulation improves erectile function of diabetic rats.
abstract: OBJECTIVE
  Promoter 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.
  
  
  METHODS
  An 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.
  
  
  RESULTS
  Adenovirus 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.
  
  
  CONCLUSIONS
  Results 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.
authors.affiliation: Department of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, People's Republic of China.
authors.forename: Tao
authors.initials: T
authors.lastname: Wang
authors.forename: Mingchao
authors.initials: M
authors.lastname: Li
authors.forename: Huixin
authors.initials: H
authors.lastname: Yuan
authors.forename: Yin
authors.initials: Y
authors.lastname: Zhan
authors.forename: Hua
authors.initials: H
authors.lastname: Xu
authors.forename: Shaogang
authors.initials: S
authors.lastname: Wang
authors.forename: Weiming
authors.initials: W
authors.lastname: Yang
authors.forename: Jihong
authors.initials: J
authors.lastname: Liu
authors.forename: Zhangqun
authors.initials: Z
authors.lastname: Ye
authors.forename: Long-Cheng
authors.initials: LC
authors.lastname: Li
mesh_terms: D000256:Adenoviridae; D000704:Analysis of Variance; D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D006152:Cyclic GMP / Q000502:physiology; D003921:Diabetes Mellitus, Experimental; D004797:Enzyme-Linked Immunosorbent Assay; D008297:Male; D009130:Muscle, Smooth / Q000502:physiology; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism*; D010410:Penile Erection / Q000502:physiology*; D012313:RNA / Q000494:pharmacology*; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction; D017423:Sequence Analysis, RNA; D015398:Signal Transduction; D015854:Up-Regulation
chemical_list: D012313:RNA; D052247:Nitric Oxide Synthase Type II; D006152:Cyclic GMP
keywords: CSMC; ED; ICP; MOI; NO; NO synthase; NOS; PCR; PDE5; RNA; RNA activation; RNAa; RT-PCR; STZ; TSS; cGMP; cavernous smooth muscle cell; cyclic guanosine monophosphate; diabetes mellitus, experimental; double stranded; ds; eNOS; endothelial NOS; erectile dysfunction; iNOS; inducible NOS; intracavernous pressure; multiplicity of infection; nNOS; neuronal NOS; nitric oxide; nitric oxide synthase type II; p-; penis; phosphodiesterase type 5; phosphorylated; polymerase chain reaction; reverse transcriptase-PCR; saRNA; shRNA; short hairpin RNA; small activating RNA; streptozocin; transcription start site