腺病毒介导的小激活RNA基因治疗对老龄大鼠阴茎勃起功能的改善

发布时间：2018-01-25 06:31

本文关键词： 勃起功能障碍基因治疗 RNA激活 iNOS 老龄大鼠　出处：《华中科技大学学报(医学版)》2017年05期 　论文类型：期刊论文

【摘要】：目的探讨靶向大鼠iNOS基因的shRNA重组腺病毒载体对老龄大鼠阴茎海绵体iNOS基因的激活作用,为勃起功能障碍(ED)的基因治疗提供实验依据。方法 24月龄SD大鼠30只随机分为3组,注射重组腺病毒Ad5-iNOSshRNA-EGFP(AdU6/shiNOS)组10只,注射对照病毒(AdU6/shControl)组10只,空白对照组10只注射PBS。注射1周后采用电刺激下检测阴茎海绵体压力和平均动脉压比值(ICP/MAP)的方法评估阴茎勃起功能,收集阴茎海绵体组织,采用real-time PCR半定量检测iNOS mRNA的表达,Western blot检测iNOS蛋白表达。结果在2.5、5.0和7.5V电压刺激时,AdU6/shiNOS组大鼠ICP/MAP均高于AdU6/shControl组和空白对照组,差异均有统计学意义(均P0.05);注射AdU6/shiNOS组海绵体组织中iNOS的mRNA和蛋白表达水平均明显高于AdU6/shControl组和空白对照组(均P0.05)。结论利用腺病毒介导的RNA激活技术能提高海绵体iNOS基因表达,改善老龄大鼠的勃起功能。
[Abstract]:Objective to investigate the effect of shRNA recombinant adenovirus vector targeting rat iNOS gene on the activation of iNOS gene in penis cavernosum of aged rats. Methods Thirty SD rats aged 24 months were randomly divided into 3 groups. Ten rats were injected with recombinant adenovirus Ad5-iNOSshRNA-EGFPN AdU6 / shiNOSand 10 in the control group (AdU6 / shControl). The penile erectile function was evaluated by measuring the ratio of cavernosum pressure to mean arterial pressure (ICP / MAPP) after 1 week of PBSs injection in the blank control group. The penile cavernous tissue was collected and the expression of iNOS mRNA was detected by real-time PCR. The expression of iNOS protein was detected by Western blot. The ICP/MAP of AdU6/shiNOS group was significantly higher than that of AdU6/shControl group and blank control group (P 0.05). The expression of mRNA and protein of iNOS in cavernous tissue of AdU6/shiNOS group was significantly higher than that of AdU6/shControl group and blank control group (all P 0.05). Conclusion Adenovirus-mediated RNA activation can enhance the expression of iNOS gene in cavernous body. To improve erectile function in aged rats.
【作者单位】：华中科技大学同济医学院附属同济医院泌尿外科;
【基金】：国家自然科学基金资助项目(No.30872574) 华中科技大学自主创新研究基金资助项目(No.0118540280) 湖北省自然科学基金资助项目(No.2017CFB638)
【分类号】：R698.1
【正文快照】： 随着中国老龄化社会的到来,勃起功能障碍(e-rectile dysfunction,ED)发病率随着老龄人口的增加而升高。美国马萨诸塞州男性老龄化研究(MMAS)指出,在随机抽取的40~70岁调查者中ED的发病率高达52%[1]。目前ED的基因治疗主要通过转入一氧化氮合酶(nitric oxide synthase,NOS)基因

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