| BACKGROUND&OBJECTIVERadical prostatectomy(RP)is the optimal treatment for patients with localized prostate cancer.The incidence of erectile dysfunction(ED)post-operation remains as high as 30 to 87%o Simultaneously,the pathophysiology of RP-ED has not been well elucidated,and the low efficacy of traditional PDE5i treatment remained a major complain in contemporary clinic.Phenotype transformation of smooth muscle cells is defined as the ability to transform from the contractile phenotype into synthetic one reversibly.Our previous studies have confirmed corpus cavernosum smooth muscle cells underwent phenotype transformation in diabetic rats,which is closely related to ED.Myocardin is a remarkably potent transcriptional coactivator and the founding member of a class of muscle transcription factors expressed exclusively in cardiomyocytes and SMCs.Our previous studies confirmed that Myocardin gene therapy can improve erectile function in diabetic ratsThis study aims to make sure the relationship between phenotype transformation and ccsmc of BCNI rats by establishing bilateral cavernous nerve injury(BCNI)rats,thus improve erectile function of BCNI rats by Myocardin gene therapy.METHODS1?36 rats were randomly assigned to BCNI or negative control(NC)groups.subgroups of 3d,5d,7d post operation were made with 6 rats each.All ratsexperienced ICP and MAP tests and the penile shaft Was removed for western blot and histochemical analysis which were used to determine the expression levels of a-sma?calponin?Myocardin and OPN,and assess the content of CCSM by HE&Masson's trichrome staining2?30 rats were randomly divided into 3 equal groups,namely NC+PBS,BCNI+Ad-Myocardin,BCNI+Ad-vector.A single intracavernous injection of 50?L PBS,Ad-Myocardin or adenovirus with empty vector were given to the corresponding rats 21 days later,all animals were anesthetized for ICP and MAP measurement and were killed thereafter.The penises were harvested,weighed,and processed for HE&Masson's trichrome staining and immunochemistry(IHC)RESULTSPart?:The 3 subgroups of 3,5,7 days in BCNI rats showed a remarkable decline in the maximum ICP compared with the NC group.?ICP/MAP in BCNI groups was significantly lower than the NC group.There was no differences in ICP among NC groups,as well as in MAP among all the BCNI and NC groups.Western blotting revealed a markedly decrease of Myocd,a-SMA and Calponin,along with elevated OPN expression at day 5 after modeling,but HE and Masson's staining revealed no morphological or SM-to-collagen.ratio changes within 7 days in the surgical groups and the controls.Part II:BCNI+Ad-Myocd rats showed partial recovery of the maximum ICP in at 21th day compared with the BCNI+Ad-vector group.However,the ?ICP/MAP in gene therapy rats was still lower than the control group.Simultaneously,HE stain revealed thinner smooth muscle layer,disordered and discontinuous cavernous sinuses in two CN injured rats.In masson's staining,SM-to-Collagen ratio was descending in both BCNI groups,with more significant reduction in the vector rats(p<0.05).Moreover,IHC suggested considerably greater expression of Myocd and a-SMA in NC+PBS and BCNI+Ad-Myocd groups under 100X and amplified 400X microscopy fields.21 days after injection,Myocd and contractile smooth muscle markers a-SMA,Calponin expression in gene-transferred group were retained as no difference to the control rats and were significantly higher than the BCNI+Ad-vector group.The CCSMC synthetic marker OPN was highly expressed in BCNI+Ad-vector rats than other groupsCONCLUSION1.Erectile dysfunction but not histological changes of BCNI rats within 7 days2.Phenotypic modulation of BCNI rat CCSMCs at day 53.Improvement of Erectile function in gene-transferred rats4.Myocardin gene therapy maintained the contractile phenotype... |
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