尿源干细胞治疗1型糖尿病性勃起功能障碍大鼠的实验研究

Effects of urine derived stem cells in the treatment of type Ⅰ diabetic erectile dysfunction rats

目的:验证尿源干细胞(USCs)治疗1型糖尿病性勃起功能障碍(DED)大鼠的效果及探索可能机制。方法:予10周龄SD雄性大鼠腹腔注射链脲佐菌素(40 mg/kg)以构建糖尿病大鼠模型,8周后行阿扑吗啡(APO)颈部皮下注射筛选DED大鼠模型。随后分离、培养、鉴定USCs。将所有大鼠分为正常对照组(n=6)、PBS组(n=6)和USCs治疗组(n=6),其中PBS组和USCs治疗组均为DED大鼠。分别将PBS或USCs(1×106/只)移植至PBS组和USCs治疗组大鼠阴茎海绵体。移植4周后分别比较三组大鼠的体重和血糖,并测定平均颈动脉内压(MAP)和海绵体内压(ICP)以评价阴茎勃起功能,分析其阴茎局部血管周细胞标志物、平滑肌/胶原比例、VEGF/VEGF/eNOS。结果:1型DED大鼠的成模率为75%。经流式分析证实USCs表达间充质干细胞表面标志物,并至少可向成骨、成脂方向分化。USCs移植4周后,除正常对照组大鼠体重保持增长外(P<0.05),PBS组、USCs治疗组的体重无明显变化(P>0.05)。此外三组大鼠治疗前后血糖均无显著变化(P>0.05)。与PBS组比较,USCs治疗组的ICP及ICP/MAP均得到显著恢复(P<0.01)。免疫荧光证实1型DED大鼠的VEGF/VEGFR1/eNOS通路和阴茎海绵窦周细胞功能(CD146)均受损,经USCs治疗4周后,该通路及CD146均得到显著恢复,平滑肌/胶原比例也得到有效恢复。结论:USCs可有效治疗1型DED大鼠,其作用机制可能是通过VEGF/VEGFR1/eNOS通路改善阴茎海绵窦内皮功能实现的。

Objective：To demonstrate the effects of urine derived stem cells（USCs）in the treatment of type Ⅰ diabetic erectile dysfunction（DED）rats and possible mechanism.Method：Twenty 10-week-old SD rats were induced to the diabetic rats by intraperitoneal injection with streptozotocin（STZ,40 mg/kg）.Type I DED rats were screened with subcutaneous injection of apomorphine.And then human urine derived stem cells（USCs）were collected from 3 healthy donors.Eighteen SD male rats were divided into 3 groups.A control group of normal rats（n=6）,and two other test groups of type 1 DED rats：PBS as a negative control（n=6）,USCs（n=6）.Totally 1×106 USCs in 100μl PBS per rats or 100 ul PBS only were injected directly into the corpora cavernosa.The body weight and blood sugar level were compared at initial and 28 days post injection.Mean arterial pressure（MAP）,intracavernosal pressure（ICP）,expression of pericytes markers（CD146）,smooth muscle/collagen ratio,and VEGF/VEGF/eNOS expression were assessed for each group.Result：After 8 weeks of STZ injection,75% rats were developed to be type Ⅰ DED rats.Cultured USCs expressed mesenchymal stem cell markers,were able to be induced into osteogenic,adipogenic lineages.No significant change of blood sugar level after implantation.Implanted USCs displayed a significant raise in ICP and ICP/MAP ratio（P〈0.01）28 days after intracavernous injection.Histological analysis demonstrated an increased expression of pericytes markers（CD146）,smooth muscle/collagen ratio,and VEGF/VEGF/eNOS expression within the cavernous tissue following USCs injection.Conclusion：The paracrine effect of USCs improved the erectile function in type Ⅰ diabetic rats by ameliorating the impaired VEGF/VEGFR1/eNOS pathway and increasing the pericytes expression and contents of smooth muscle.