hTERT修饰ADSCs移植治疗老年性ED的作用及其机制研究

Aging has been generally recognized as a high-risk factor for ED,Phosphodiesterase V inhibitors (PDE5I) have poor efficacy for organic ED.The preliminary study found that aged cavernosal tissue shows decrease in SMC, SMC life can be extended by hTERT transfected. Stem cell can be directly differentiated to replace damaged cells and be used as gene therapy vectors. The adipose stem cells (ADSCs) for ED treatment were be taken seriously. We speculate that hTERT modification could improve the proliferative capacity of ADSCs, and ADSCs could transfect hTERT to the corpus cavernosum as a carrier, the SMC number and function could be improved. This project, for the first time,ADSCs is modified with hTERT and be transplanted to the cavernosum of aged rats.Erectile function and ultrastructure of penis are observed. By RT-PCR,Western,immunofluorescence and flow cytometry,ADSCs' proliferation,apoptosis, and tumorigenicity and differentiation potential,SMC, endothelial cells, growth factors,the expression of hTERT and NOS are detected.To explore the role and its regulatory mechanism of the hTERT gene-modified ADSCs transplantation in the treatment of elderly ED. Provide new strategies for the prevention of aging ED.

衰老是勃起功能障碍(ED)的高危影响因素。磷酸二酯酶V型抑制剂(PDE5I)对老年性ED疗效欠佳。前期研究发现：老年性ED阴茎平滑肌细胞(SMC)减少,端粒酶催化亚单位(hTERT)转染可使SMC寿命延长。干细胞移植可直接分化取代受损细胞及作为基因治疗的载体。脂肪干细胞(ADSCs)用于ED治疗受到重视。我们推测:hTERT修饰可提高ADSCs的增殖能力，ADSCs作为载体可转染hTERT至海绵体，使SMC数量及功能得到改善。本项目拟首次以hTERT修饰ADSCs并移植老年大鼠海绵体, 检测阴茎勃起功能,超微结构,通过RT-PCR、Western、免疫荧光、流式细胞仪等测定ADSCs增殖、凋亡、致瘤性及分化潜能，SMC、内皮细胞数量,生长因子,hTERT、NOS 等表达。阐明衰老对ADSCs的影响，探讨hTERT基因修饰ADSCs移植治疗老年ED的作用及其机制。为防治老年性ED提供新策略。

勃起功能障碍（ErectileDysfunction，ED）是泌尿男性科最常见的疾病之一，阴茎海绵体平滑肌细胞（SMC）舒张是阴茎勃起的关键。本研究首次探讨hTERT基因修饰ADSCs自体移植治疗老年ED的作用及其机制。检测并探讨了hTERT修饰提高ADSCs增殖能力、分化潜能的机制，进行了转染鉴定、细胞增殖、凋亡检测、致瘤性检测、细胞因子检测。研究了hTERT基因修饰ADSCs自体移植治疗老年ED的作用及其机制：hTERT基因修饰ADSCs自体移植老年大鼠阴茎组织后，检测了阴茎组织增殖及凋亡、免疫荧光双染法评价脂肪源干细胞向平滑肌样细胞、内皮细胞分化、检测大鼠阴茎海绵体内NOS的表达和活性、细胞因子检测、检测hTERT表达、海绵体内cGMP、cAMP浓度测定、检测阴茎海绵体是否存在免疫和炎症反应。研究结果提示：hTERT修饰的ADSCs后，ADSCs细胞增殖能力增强，凋亡减少，分化能力增强。ADSCs自体移植可预防老年性ED。