MSCs-exosomes介导miR-210调控PI3K/AKT/Nrf2通路对糖尿病性勃起功能障碍的作用及机制研究

The transplantation of bone marrow mesenchymal stem cells (MSCs) in the treatment of diabetic erectile dysfunction (DMED) has the shortcoming of tumorigenicity and short curative effect. Therefore, acellular therapy is the research direction at present. Recent focus on this issue is the application of exosomes. Our previous studies have found that the dysfunction of vascular endothelial cells and smooth muscle cells caused by the increased oxidative stress may be the main pathogenesis of DMED. We also found activation of Nrf2 gene could significantly improve erectile function of diabetic rats. Recent studies have found that miR-210 mediated by MSCs-exosomes can improve the local oxidative stress microenvironment, which is presumably related to activation of PI3K/AKT pathway. Up to now, there are no reports on the mechanism of miR-210 treatment for DMED. Therefore, we propose that local injection of miR-210 mediated by MSCs-exosomes is more effective in the treatment of DMED. The study intends to study the change of PI3K/AKT/Nrf2 pathway. In vivo and in vitro experiments will be combined according to the molecular-cell-organism level. We will explore the effect of miR-210 on the oxidative stress state of cavernous body and the changes of vascular endothelial cell function and smooth muscle cell function in the cavernosum. The study is aimed to explore a new acellular therapy for DMED.

骨髓间充质干细胞（MSCs）移植治疗糖尿病性勃起功能障碍（DMED）有致瘤性、疗效短等缺点，因而无细胞治疗是目前研究热点。近期多聚焦于外泌体（exosome）应用。我们前期发现：增强的氧化应激状态及导致的血管内皮细胞和平滑肌细胞功能不全是DMED主要原因；激活Nrf2基因能改善糖尿病大鼠勃起功能。最新研究发现MSCs-exosomes介导miR-210能改善受体局部氧化应激水平，推测与激活PI3K/AKT有关。国内外尚未有miR-210治疗DMED的报道。因此，我们提出局部注射MSCs-exosomes介导miR-210治疗DMED的假设。本项目拟以PI3K/AKT/Nrf2通路改变为切入点，按照分子-细胞-整体水平，体内、外实验结合的方法，探究miR-210对糖尿病性阴茎海绵体氧化应激状态的作用，以及阴茎海绵体血管内皮细胞功能、平滑肌细胞功能变化情况，探索DMED无细胞治疗的新思路。

干细胞移植治疗糖尿病性勃起功能障碍（DMED）有致瘤性、疗效短等缺点，因而无细胞治疗是目前研究热点。近期多聚焦于外泌体（exosome）的应用。我们的研究通过体内及体外试验证实了骨髓间充质干细胞（MSCs）来源的外泌体介导miR-210通过调控PI3K/AKT/Nrf-2信号通路减轻糖尿病大鼠氧化应激，减少平滑肌细胞及内皮细胞凋亡，从而改善勃起功能。在研究过程中，我们发现单纯外泌体进行海绵体注射存在“突释”现象，使得外泌体留存时间减少，限制了其应用，因此我们创新性研发了一种新型水凝胶 (HG) ，并耦合干细胞来源外泌体(HG@Exo) 来治疗CNI-ED。我们的研究证明，HG 表现出出色的可注射性、结构稳定性和温敏性，可以显著延长外泌体的留存时间并将其缓慢释放。我们还观察到，将HG@Exo应用于神经损伤部位后，CNI-ED大鼠模型的勃起功能得到显著改善。我们的研究表明外泌体治疗是一种安全有效的治疗 DMED 新方法，miR-210可能是DMED新的治疗靶点，我们的研究将有助于摆脱依赖于干细胞的细胞治疗，进入无细胞治疗的时代。而且我们开发的新型水凝胶耦合外泌体的应用为提高勃起功能障碍的治疗效果提供了一个创新性且有临床意义的方向，具有广阔的应用前景。