摘要
目的 探索在海绵体神经损伤的大鼠神经性阴茎勃起功能障碍（NED）模型中,注射BDNF过表达的MSCs在神经再生和修复中的作用.方法 建立脑源性神经营养因子（BDNF）过表达的骨髓间充质干细胞（MSCs）,通过Western Blot检测BDNF和VEGF-A的表达的水平.通过Elisa检测BDNF和VEGF-A在细胞上清液中的含量.取120只3月龄大鼠随机分为4组：第1组为假手术组（Sham组）,其余三组均损伤双侧海绵体神经,第2组（Control组）不再进行干预,第3组（MSC组）和第4组（BDNF-MSC）分别在关腹前注射MSCs和BDNF过表达的MSCs.大鼠在手术造模后12周,通过海绵体神经电刺激并测阴茎海绵体内压（ICP）来评价阴茎勃起功能.取大鼠阴茎海绵体组织行免疫组化评价诱导型NO合酶（iNOS）的表达水平.结果 通过绿色免疫荧光试验证实BDNF过表达MSCs制备成功.Western Blot验证结果显示BDNF过表达的MSC细胞中的BDNF和VEGF-A表达水平较MSC细胞明显上调,ELISA验证结果显示BDNF过表达的MSC细胞上清液中的BDNF和VEGF-A表达水平较MSC细胞上清液明显上调.手术后12周,通过海绵体神经电刺激并测ICP来评价阴茎勃起功能,其中假手术组平均最大ICP为（102.20±15.61）cmH2O,实验对照组为（35.47±13.16）,MSC干预组为（43.79±16.94）cmH2O,BDNF-MSC干预组为（66.04±15.33） cmH2O.免疫组化结果显示,BDNF-MSC干预组大鼠阴茎海绵体组织内iNOS阳性表达细胞数明显高于MSC干预组.结论 本研究结果显示局部注射MSCs和BDNF过表达的MSCs对于大鼠的NED具有治疗作用.
Objective To investigate the effects of intracavernosal injection with brain-derived neurotrophic factor （BDNF） overexpressed mesenchymal stem cells （BDNF-MSCs） on recovery of erectile function after cavernous nerve injury （CNI）. Methods Stable MSCs with BDNF over epression were first established. Western blot and ELISA were applied to detect the levels of BDNF and Vascular endothelial growth factor A （VEGF-A） in cells or their supernatants respectively. A total of 120 3-month-old Sprague-Dawley （SD） rats were used in the study and they were divided randomly into four groups. Rats in sham-operated group were treated with a sham operation without cavernous nerve manipulation. Rats in the other three groups were treated with bilateral CNI,but rats in the control group had no further manipulation. Rats in MSC and BDNF-MSC groups were treated by intracavernosal injection with MSCs or BDNF-MSCs respectively. Erectile function was assessed by cavernosal nerve electrostimulation 12 weeks after operation. The penile tissues werethen collected for analysis of inducible nitric oxide synthase （iNOS） level. Results BDNF-MSC cells were successfully established. Western blot and ELISA results demonstrated that both BDNF and VEGF-A were all increased in the BDNF- MSC cells and its supernatant, as compared with that in MSC cells. After nerve crushing, the erectile functional evaluation of rats in the control group at 3 months showed a lower mean maximal intracavernous pressure （ICP） under CN stimulation, at （35.47±13.16）cmH20, than that of rats in the sham group, at （102.20±15.61）cmH20. Meanwhile, the [CP of BDNF-MSC group was significantly higher than that of the controls, at （66.04±15.33）cmH20. lmmunohistochemical staining showed an increase of iNOS in penile smooth muscle cells of BDNF-MSC group. Conclusion intracavernous injection with BDNF- overexpressed MSCs may enhance the therapeutic effect of M SCs for the treatment of erectile dysfunction.
出处
《中国男科学杂志》
CAS
CSCD
2015年第4期3-8,共6页
Chinese Journal of Andrology
基金
中央级公益性科研院所基本科研业务费资助(编号:GY2012G-3)
关键词
勃起功能障碍
间质干细胞
脑源性神经营养因子
erectile dysfunction
mesenchymal stem cell
brain-derived neurotrophic factor
