转染胰岛素样生长因子-1基因对原代培养的阴茎海绵体平滑肌细胞增殖的影响  

Gene transfer of insulin-like growth factor-1 promotes the proliferation of corpus cavernosal smooth muscle cells in vitro in rats

作　　者：蒲小勇[1,2] 闻安民[1] 刘久敏[2] 郑祥光[2] 王怀鹏[2] 徐战平[2] 张仁礼[1] 周香雪 

机构地区：[1]广东省人民医院生殖中心,广州510080 [2]广东省人民医院泌尿外科,广州510080 [3]山大学附属第一医院黄埔院区神经内科

出　　处：《中华实验外科杂志》2012年第8期1577-1579,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30801147、30872572）;中央高校基本科研业务费专项基金资助项目（09ykpy16）;中国博士后科学基金资助项目（20060400779）;广东省自然科学基金资助项目（07001203）;广东省医学科研基金资助项目（A2008033、B2010094）

摘　　要：目的观察转染胰岛素样生长因子-1（IGF-1）基因对原代培养的阴茎海绵体平滑肌细胞（CCSMCs）增殖的影响，探讨IGF-1基因治疗勃起功能障碍（ED）的可能机制。方法构建pcDNA3．1-IGF-1，体外原代培养大鼠CCSMCs并免疫组织化学染色鉴定；CCSMCs分3组：转染pcDNA3．1-IGF-1组、pcDNA3．1组和对照组，转染后不同时间段（3、5、7、9d）噻唑蓝（Mrrr）比色法检测细胞增殖，同时不同时间段（24、48、96h）酶免疫分析法（EIA）定量检测CCSMCs分泌IGF-1含量的变化。结果成功克隆pcDNA3．1-IGF-1载体，成功原代培养CCSMCs；转染pcDNA3．1．1GF-1组细胞在转染后5、7、9d（分别为0．72±O．08、0．80±0．06、0．82±0．07）较转染pcDNA3．1组（分别为0．58±0．07、0．63±0．05、0．6l±0．08）和对照组（0．59±0．06、0．61±0．07、0．62±0．03）增殖明显增加，呈时间依赖性，差异有统计学意义（P〈0．05）。与转染pcDNA3．1组[分别为（46±7）、（48±9）、（50±11）μg/L]和对照组[分别为（47±11）、（52±10）、（51±10）μg/L]比较，转染pcDNA3．1-IGF-1组细胞在转染后24、48、96hCCSMCs分泌的IGF-1含量[分别为（58±8）、（69±11）、（71±8）μg/L]增加，差异有统计学意义（P〈0．05）。结论转染IGF-1基因可提高CCSMCs的增殖能力和增加IGF-1的分泌，可能是IGF-1基因治疗ED的基础。Objective To study whether gene transfer of insulin-like growth factor-1 （ IGF-1 ） can promote the proliferation of corpus cavernosal smooth muscle ceils （CCSMCs） in rats and explore the possi- ble mechanism of IGF-1 gene therapy for erectile dysfunction （ ED ）. Methods The pcDNA3.1-IGF-1 vector was constructed and sequenced. The CCSMCs were subjected to primary culture and identified, and divided into 3 groups： pcDNA3. 1-IGF-1 group, pcDNA3. 1 group （vehicle-only） and control group. The cell proliferation was measured by using methyhhiazoletetrazolium （MTT） assay in the three cell strains at 3rd, 5th, 7th and 9th day after transfection. The enzyme immunoassay （EIA） kit was used to quantify IGF-I levels in in the three cell strains at 24, 48 and 96 h after transfection. Results The pcDNA3.1- IGF-1 vector was successfully cloned with correct open reading fragment. The CCSMCs were confirmed by an- ti-or smooth muscle actin antibodies. Gene transfer of IGF-1 significantly promoted proliferation of CCSMCs in a time-dependent manner at 5th, 7th and 9th day （A570 ： 0. 72 ±0. 08, 0. 80 ±0. 06, 0. 82 ±0. 07 ,P 〈0. 05, respectively）. As compared with pcDNA3. 1 group [ （46 ±7） , （48 ±9）, （50 ± 11 ） μg/L, respectively] and control group [ （47 ± 11 ）, （52± 10） , （51 ± 10）μg/L, respectively] , the secretion of IGF-1 in CCSMCs was significantly increased in pcDNA3.1-IGF-1 group [ （58± 8 ） , （69 ± 11 ）, （71 ± 8 ） μg/L, respectively ] at 24, 48 and 96 h （ all P 〈 0. 05 ）. Conclusion The gene transfer of IGF-1 promotes the proliferation of CCSMCs in rats and might contribute to the mechanism of using IGF-1 gene therapy for ED.

关 键 词：胰岛素样生长因子-1 勃起功能障碍 增殖 阴茎海绵体 平滑肌细胞 

分 类 号：R698.1[医药卫生—泌尿科学]