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[尿源性干细胞对海绵体神经损伤大鼠勃起功能障碍的保护作用]

[Protective effect of urine-derived stem cells on erectile dysfunction in rats with cavernous nerve injury].

作者信息

Chen Wan-Mei, Yang Qi-Yun, Bian Jun, Han Da-Yu, Lai De-Hui, Sun Xiang-Zhou, Deng Chun-Hua

机构信息

Department of Anesthesiology, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510080, China.

Department of Urology, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510080, China.

出版信息

Zhonghua Nan Ke Xue. 2018 Jun;24(6):483-490.

Abstract

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 1×106/200 μl 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 α-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 treatment, the rats in the USC group, as compared with those in the PBS and BCNI model control groups, showed significant increases in the mICP ([81 ± 9.9] vs [31 ± 8.3] and [33 ± 4.2] mmHg, P <0.05), mICP/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 ± 4.92]%, P <0.05), the eNOS/GAPDH ratio (0.52 ± 0.08 vs 0.31 ± 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 and 0.38 ± 0.04, P <0.05), but a remarkable decrease in the collagen I/GAPDH ratio (0.28 ± 0.06 vs 0.68 ± 0.04 and 0.70 ± 0.10, P <0.05). The ratio of smooth muscle to collagen in the corpus cavernosum was significantly higher in the USC than in the PBS and BCNI model control groups (17.91 ± 2.86 vs 7.70 ± 3.12 and 8.21 ± 3.83, P <0.05) while the rate of cell apoptosis markedly lower in the former than in the latter two (3.31 ± 0.83 vs 9.82 ± 0.76, P <0.01; 3.31 ± 0.83 vs 9.75 ± 0.91, P <0.05).

CONCLUSIONS

Intracavernous injection of USCs can protect the erectile function of the rat with cavernous nerve injury by protecting the nerves, improving the endothelial function, alleviating fibrosis and inhibiting cell apoptosis in the cavernous tissue.

摘要

目的

探讨人尿源干细胞(USCs)对海绵体神经损伤(CNI)大鼠勃起功能和海绵体结构的保护作用。

方法

将60只具有正常性功能的成年雄性SD大鼠随机分为数量相等的四组:假手术组、双侧CNI(BCNI)模型对照组、磷酸盐缓冲盐水(PBS)组和USCs组。后三组大鼠通过钳夹双侧海绵体神经建立BCNI模型。建模后,PBS组和USCs组大鼠分别通过海绵体内注射200 μl PBS和1×106/200 μl PBS的USCs进行治疗,持续28天。然后,通过电刺激盆神经节计算大鼠的最大海绵体内压(mICP)以及mICP与平均动脉压的比值(mICP/MAP),通过免疫组织化学染色确定阴茎背神经总面积中nNOS或NF200阳性神经纤维的比例,通过蛋白质免疫印迹法检测内皮细胞标志物eNOS、平滑肌标志物α-SMA和I型胶原蛋白的水平,分别通过Masson染色和TUNEL法测量海绵体中平滑肌与胶原蛋白的比例以及细胞凋亡率。

结果

治疗28天后,与PBS组和BCNI模型对照组相比,USCs组大鼠的mICP([81 ± 9.9] vs [31 ± 8.3]和[33 ± 4.2] mmHg,P <0.05)、mICP/MAP比值(0.72 ± 0.05 vs 0.36 ± 0.03和0.35 ± 0.04,P <0.05)、阴茎背神经总面积中nNOS阳性神经纤维的比例([11.31 ± 4.22]% vs [6.86 ± 3.08]%和[7.29 ± 4.84]%,P <0.05)以及NF200阳性神经纤维的比例([27.31 ± 3.12]% vs [17.38 ± 2.87]%和[19.49 ± 4.92]%,P <0.05)、eNOS/GAPDH比值(0.52 ± 0.08 vs 0.31 ± 0.06和0.33 ± 0.07,P <0.05)以及α-SMA/GAPDH比值(1.01 ± 0.09 vs 0.36 ± 0.05和0.38 ± 0.04,P <0.05)均显著升高,但I型胶原蛋白/GAPDH比值显著降低(0.28 ± 0.06 vs 0.68 ± 0.04和0.70 ± 0.10,P <0.05)。USCs组海绵体中平滑肌与胶原蛋白的比例显著高于PBS组和BCNI模型对照组(17.91 ± 2.86 vs 7.70 ± 3.12和8.21 ± 3.83,P <0.05),而细胞凋亡率则显著低于后两组(3.31 ± 0.83 vs 9.82 ± 0.76,P <0.01;3.31 ± 0.83 vs 9.75 ± 0.91,P <0.05)。

结论

海绵体内注射USCs可通过保护神经、改善内皮功能、减轻纤维化和抑制海绵体组织细胞凋亡来保护海绵体神经损伤大鼠的勃起功能。

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