Institute of Urinary Surgery, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
Institute of Urinary Surgery, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
Biomed Pharmacother. 2017 Aug;92:595-605. doi: 10.1016/j.biopha.2017.04.088. Epub 2017 Jun 1.
Erectile dysfunction (ED) is considered to be incapable of obtaining or/and keeping a sufficient erection function to receive the satisfactory during the sexual intercourse. This study aims to investigate the effects of telomerase reverse transcriptase (hTERT) modified adipose tissue derived stem cells (ADSCs) autologously injected into cavernosa of the ED rats on erectile function. The ADSCs were isolated form the rat subcutaneous adipose tissue sample, and identified by examining the CD29 and CD44 molecule. The ED model was established by using 100μg/kg apomorphine (APO). The adenovirus expressing rat hTERT (Ade-hTERT vector) was established, and transfected into ADSCs and injected into ED rat model, respectively. Telomerase activity, cell growth, cell apoptosis were analyzed by using TRAP ELISA assay, CCK8 assay and flow cytometry assay, respectively. The trophic growth factors were examined by using enzyme-linked immunosorbent assay (ELISA). The mRNA and proteins were detected by using semi-quantitative PCR and western blot assay, respectively. Ade-hTERT vector was highly expressed in both ADSCs and ED rat mode. The hTERT expression enhanced the telomerase activity, inhibits cell apoptosis and enhances proliferation of ADSCs (P<0.05). hTERT expression triggers the secretory function of ADSCs and induces differentiative potential of ADSCs. hTERT expression inhibits apoptosis and increases eNOS and nNOS levels in older ED rats compared to the Ade-vector injected ED rats (P<0.05). In conclusion, the hTERT modification could enhance the ADSCs proliferation, and hTERT modified ADSCs could increase the anti-oxidative stress capacity in the ED rat model.
勃起功能障碍(ED)被认为是无法获得和/或保持足够的勃起功能,以在性交中获得满意的效果。本研究旨在探讨端粒酶逆转录酶(hTERT)修饰的脂肪组织来源干细胞(ADSCs)自体注射到 ED 大鼠海绵体内对勃起功能的影响。ADSCs 从大鼠皮下脂肪组织样本中分离出来,并通过检测 CD29 和 CD44 分子进行鉴定。使用 100μg/kg 阿扑吗啡(APO)建立 ED 模型。建立了表达大鼠 hTERT 的腺病毒(Ade-hTERT 载体),并分别转染到 ADSCs 中,然后注射到 ED 大鼠模型中。通过 TRAP ELISA 测定、CCK8 测定和流式细胞术分别分析端粒酶活性、细胞生长和细胞凋亡。采用酶联免疫吸附试验(ELISA)检测营养生长因子。通过半定量 PCR 和 Western blot 检测分别检测 mRNA 和蛋白质。Ade-hTERT 载体在 ADSCs 和 ED 大鼠模型中均高度表达。hTERT 表达增强了端粒酶活性,抑制了细胞凋亡,促进了 ADSCs 的增殖(P<0.05)。hTERT 表达触发了 ADSCs 的分泌功能,并诱导了 ADSCs 的分化潜能。与注射 Ade-vector 的 ED 大鼠相比,hTERT 表达抑制了老年 ED 大鼠的细胞凋亡,并增加了 eNOS 和 nNOS 水平(P<0.05)。总之,hTERT 修饰可以增强 ADSCs 的增殖,hTERT 修饰的 ADSCs 可以增加 ED 大鼠模型的抗氧化应激能力。