腺病毒介导PDE5A_1反义核酸改善糖尿病兔阴茎勃起功能的实验研究  

Adenoviral gene transfer of antisense cDNA of the human PDE5A_1 promoter gene to penis enhances erectile function in rabbits with diabetes mellitus

作　　者：赵永平[1] 白文俊[1] 陈国强[1] 刘士军[1] 邓庆平[1] 孔雷[1] 闫征[1] 王晓峰[1] 

机构地区：[1]北京大学人民医院泌尿外科,100044

出　　处：《中华泌尿外科杂志》2005年第8期562-565,共4页Chinese Journal of Urology

基　　金：国家自然科学基金项目资助(30371419)

摘　　要：目的探讨腺病毒介导PDE5A1反义核酸对海绵体平滑肌PDE5基因表达的抑制作用及对兔阴茎勃起功能的影响。方法25只成年雄性新西兰兔随机分为5组:1组为正常对照组,4组静脉注射四氧嘧啶建立糖尿病性勃起功能障碍(ED)动物模型,分别将携带反义基因腺病毒载体、携带β-半乳糖苷酶基因腺病毒载体(β-gal组)、空载体及0.9%NaCl转染模型组阴茎海绵体。转染后第7天,电刺激盆神经测定各组阴茎海绵体内压(NSICP),评价阴茎勃起功能,ELISA法测定海绵体组织cGMP浓度,Westernblot法分析海绵体组织中β-gal表达水平。结果反义核酸组海绵体组织cGMP浓度(25.6±2.5)fmol/mg蛋白,明显高于β-gal组(8.8±0.9)fmol/mg蛋白、空载体组(8.3±1.1)fmol/mg蛋白和糖尿病对照组(7.4±0.8)fmol/mg蛋白,差异有统计学意义(P<0.05);反义核酸组NSICP增加幅度(66.2±3.6)mmHg,明显高于β-gal组(38.2±2.5)mmHg、空载体组(35.2±2.2)mmHg及糖尿病对照组(36.6±2.7)mmHg,差异有统计学意义(P<0.05);与正常对照组(65.2±3.2)mmHg比较差异无统计学意义(P>0.05);β-gal组海绵体组织β-gal表达水平明显高于其他组。结论腺病毒介导PDE5A1反义基因能改善糖尿病兔阴茎勃起功能,PDE5A1可作为ED基因治疗的重要靶基因。Objective To investigate whether the adenoviral-mediated expression of the antisensee DNA of the human PDE5A1 promoter gene can improve erectile function in rabbits with diabetes mellitus （ DM）. Methods Twenty-five male rabbits was randomly divided into 5 groups as follows ：4 treatment （ n = 5 in each group） and normal control （n = 5 ） groups. Alloxan was injected intravenously into the treatment groups to establish diabetic erectile dysfunction in animal models. The pAd/CMV/V5/antisense-PDE5A1 ,pAd/CMV/V5-GW-lacZ （β-gal group）, vehicle and 0.9% NaC1 were transfected into the corpus cavernosum of the DM rabbits in the 4 treatment groups, respectively. Seven days after transfeetion, the intraeavernosal pressure during pelvic nerve stimulation （NSICP） was compared with that in normal control animals. Adenoviral transfection efficiency of β-gal reporter gene was measured by Western blot analysis, and cGMP levels in eavernosal tissue were detected by ELISA. Results Seven days after transfeetion with pAd/CMV/V5/antisense-PDE5A1 , cGMP level in cavernosal tissue （25.6±2.5 ） fmol/mg protein was significantly higher than those in β-gal group [ （ 8.8±0.9） fmol/mg protein ] , vehicle group [ （ 8.3± 1.1 ） fmol/mg protein] ,DM control group [ （7.4±0. 8） fmol/mg protein,P 〈 0. 05 ]. The increase in NSICP of the pAd/CMV/V5/antisense-PDE5A1 -transfected rabbits [ （66.2±3.6 ） mmHg ] was significantly higher thanthose of β-gal group [ （ 38.2 ± 2.5） mmHg], vehicle group [ （35.2±2.2） mmHg], and DM control group [（36.6± 2.7） mmHg ,P 〈 0.05]. There was no significant difference between the pAd/CMV/V5/anti sense-PDE5A1 group and normal control group [ （65.2± 3.2 ） mmHg, P 〉 0.05 ]. Adenoviral expression of β-gal reporter gene was significantly higher in cavernosal tissue in β-gal group than in other groups. Conclusions These results suggest that in vivo adenoviral gene transfer of antisense cDNA of the huma nPDE5A1 promoter gene to penis can improve erectil

关 键 词：腺病毒 PDE5A1反义核酸 糖尿病 阴茎勃起功能 实验研究 

分 类 号：R698.2[医药卫生—泌尿科学]