摘要
目的:探讨尿源干细胞(USC)对海绵体神经损伤性勃起功能障碍(CNIED)大鼠勃起功能和阴茎海绵体组织结构的保护作用。方法:60只成年雄性SD大鼠随机平均分为4组(n=15只/组):假手术组、双侧海绵体神经钳夹损伤组(BCNI组)、PBS组、USC组。假手术组予暴露双侧海绵体神经后直接关闭手术切口,其余3组均予血管钳钳夹双侧海绵体神经1 min,建立CNIED模型;PBS组和USC组分别予阴茎海绵体注射PBS(200μl)或USC(1×10~6细胞/200μl PBS)。治疗28 d后测定大鼠的最大海绵体内压(mICP)和m ICP/平均动脉压(mICP/MAP),并通过Western印迹检测海绵窦内皮细胞标志物e NOS,平滑肌标志物α-SMA,以及Collagen I,通过免疫组化检测海绵体阴茎背神经内的神经标志物(nNOS、NF-200),Masson染色检测海绵体平滑肌/胶原比值,以及TUNEL染色检测海绵窦内细胞凋亡水平。结果:治疗28 d后,USC组大鼠的mICP及mICP/MAP均较PBS组[(81±9.9)mm Hg vs(31±8.3)mm Hg,0.72±0.05 vs 0.36±0.03,P<0.05]和BCNI组[(81±9.9)mm Hg vs(33±4.2)mm Hg,0.72±0.05 vs 0.35±0.04,P<0.05]显著升高。免疫组化结果显示:USC组大鼠背神经内n NOS、NF-200阳性神经纤维面积的比例(%)均较PBS组(11.31±4.22 vs 6.86±3.08,27.31±3.12 vs 17.38±2.87,P<0.05)和BCNI组(11.31±4.22 vs 7.29±4.84,27.31±3.12 vs 19.49±4.92,P<0.05)显著增加;Western印迹检测结果显示:USC组大鼠e NOS/GAPDH比值较PBS组(0.52±0.08 vs 0.31±0.06,P<0.05)和BCNI组(0.52±0.08 vs 0.33±0.07,P<0.05)均显著提高,α-SMA含量亦较PBS组(1.01±0.09 vs 0.36±0.05,P<0.05)和BCNI组(1.01±0.09 vs 0.38±0.04,P<0.05)显著提高,而Collagen I含量较PBS组(0.28±0.06 vs 0.68±0.04,P<0.05)和BCNI组(0.28±0.06 vs 0.70±0.10,P<0.05)显著降低;且Masson染色结果示USC组大鼠阴茎平滑肌/胶原比值(%)亦较PBS组(17.91±2.86 vs 7.70±3.12,P<0.05)和BCNI组(17.91±2.86 vs 8.21±3.83,P<0.05)显著升高。TUNEL染色显示USC组大鼠海绵窦内细胞的凋亡指数(%)较PBS组(3.31±0.83
Objective： To investigate the protective effect of human urine-derived stem cells （USCs） on erectile function and cavernous structure in rats with cavernous nerve injury （CNI）. Methods： Sixty adult male SD rats with normal sexual function were randomly divided into four groups of equal number： sham operation, bilateral CNI （BCNI） model control, phosphate buffered saline （PBS） , and USC. The BCNI model was established in the latter three groups of rats by clamping the bilateral cavernous nerves. After modeling, the rats in the PBS and USC groups were treated by intracavernous injection of PBS at 200 μl and USCs at 1 × 10^6/200μl PBS respectively for 28 days. Then, the maximum intracavernous pressure （mICP） and the ratio of mICP to mean arterial pressure （mICP/MAP） of the rats were calculated by electrical stimulation of the major pelvic ganglions, the proportion of nNOS- or NF200- positive nerve fibers in the total area of penile dorsal nerves determined by immunohistochemical staining, the levels of endothelial cell marker eNOS, smooth muscle marker c^-SMA and collagen I detected by Western blot, and the smooth muscle to collagen ratio and the cell apoptosis rate in the corpus cavernosum measured by Masson staining and TUNEL, respectively. Results ： After 28 days of treat- ment, the rats in the USC group, as compared with those in the PBS and BCNI model control groups, showed significant increases in the mlCP （[81 ± 9.9] vs [31 ±8.3] and [33 ± 4.2] mmHg, P 〈0.05）, mlCP/MAP ratio （0.72 ± 0.05 vs 0.36 ± 0.03 and 0.35 ± 0.04, P 〈 0.05 ）, the proportions of nNOS-positive nerve fibers （ [ 11.31 ±4.22 ] % vs [ 6.86 ± 3.08 ] % and [ 7.29 ± 4.84 ] % , P 〈 0.05 ） and NF200-positive nerve fibers in the total area of penile dorsal nerves （ [ 27.31 ± 3.12 ] % vs [ 17.38 ±2.87]% and [19.49 ± 4.921%, P 〈0.05）, the eNOS/GAPDH ratio （0.52 ± 0.08vs0.31 ± 0.06 and 0.33 ± 0.07, P 〈 0.05） , and the α-SMA/GAPDH ratio （1.01 ± 0.09 vs 0.36± 0.05 andO.
作者
陈婉媚
杨其运
卞军
韩大愚
赖德辉
孙祥宙
邓春华
CHEN Wan-mei;YANG Qi-yun;BIAN Jun;HAN Da-yu;LAI De-hui;SUN Xiang-zhou;DENG Chun-hua(Department of Anesthesiology;Department of Urology, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510080, China;Department of Urology, The Third Affiliated Hospital of Southern Medi- cal University, Guangzhou, Guangdong 510630, China;Department of Urology, The Fifth Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong 510700, China)
出处
《中华男科学杂志》
CAS
CSCD
北大核心
2018年第6期483-490,共8页
National Journal of Andrology
基金
国家自然科学基金面上项目(81471449
81671449)
国家自然科学基金青年科学基金资助项目(30901487)
广东省自然科学基金(2015A030313013)
广东省自然科学基金博士启动项目(2015A030310027)~~
