超声微泡介导血管内皮生长因子基因转染大鼠阴茎组织

Ultrasound-mediated microbubble destruction enhances VEGF_(165) gene to the rat penis

目的探讨超声靶向破坏微泡介导血管内皮生长因子165(VEGF165)转染于高脂模型大鼠阴茎海绵体组织的可行性。方法以含4%胆固醇及1%胆酸饲料饲养36只2月龄雄性SD大鼠3个月,建立大鼠高脂模型,将其随机均分为高脂模型组(对照组)、VEGF165组和1.0W/cm2超声+微泡+VEGF165组(US+MB+VEGF165组)。于基因转染后7天处死大鼠,采用荧光定量PCR检测VEGF165基因表达水平,以Western Blot检测大鼠阴茎组织VEGF蛋白质的表达水平,用免疫组织化学法(IHC)检测大鼠阴茎组织内皮型一氧化氮合成酶(eNOS)蛋白质表达。结果转基因7天后,US+MB+VEGF165组VEGF165基因水平明显高于VEGF165组及对照组(P均<0.05),其阴茎海绵体组织VEGF蛋白质表达较VEGF165组及对照组增加(P均<0.05);IHC结果显示,US+MB+VEGF165组大鼠阴茎组织eNOS较其他组高表达(P均<0.05)。结论超声靶向破坏微泡可介导VEGF165基因在大鼠高脂模型阴茎海绵体组织的高效转移,为基因治疗高脂勃起功能障碍提供了实验依据。

Objective To investigate the possibility of improving the delivery of vascular endothelial growth factor 165 （VEGF165） gene to eavernosum tissue in rat models of hypercholesterolemia by using ultrasound targeted microbubble de struetion （UTMD）. Methods Thirty-six 2-month-old male Sprague-Dawley （SD） rats were included. The animals fed on the artificial diet containing 4 % cholesterol and 1% cholic acid for 3 months. All rats were randomly divided into the following group averagely after the establishment of hypercholesterolemia models： Model group alone （control group）, VEGF165 group and 1.0 W/cm^2 ultrasonic irradiation＋microbubbles＋VEGF165 group （US＋MB＋VEGF165 group）. All rats were sacrificed and the cavernosum were harvested 7 days after UTMD. The expression of VEGF165 mRNA was detec ted with real time PCR, the expression of VEGF protein were detected by Western Blot, while the expression of eNOS pro tein were evaluated with immunohistochemical examination. Results The results from real time PCR revealed that corn pared with the control group and VEGFI^5 group, the expression of VEGF mRNA markedly increased in US＋ MB＋ VEGF,65 group （both P^0.05）. Western Blot analysis showed that the expression of VEGF in US＋MB＋VEGF165 group significantly increased more than the other groups （both P〈0.05）. Immunohistochemical examination showed that the ex pression of eNOS protein in US＋MB＋VEGF165 group also significantly increased than the other groups （both P〈0.05）. Conclusion UTMD is a noninvasive method that can effectively enhance the delivery of targeted VEGF165 genes to cavernosum tissue in rat models of hypercholesterolemia, which forms the experimental base of gene therapy for erectile dysfunction.