The studies with adipose-derived stem/stromal cells therapeutic development of erectile dysfunction

简介和目的：糖尿病患者对5型磷酸二酯酶抑制剂（PDE-5I）的治疗更加难治，因为其海绵体严重受损。我们以前报道过急性肾上腺髓质素（AM）进入海绵穴体可以增强大鼠的勃起功能。在筛选脂肪组织源性干细胞（ASC）产生的细胞因子过程中，我们发现ASC产生AM，特别是当ASC在含有血管内皮细胞（VEC）生长因子的培养基中培养时。在这项研究中，我们研究了AM是否与勃起功能恢复有关。方法：从Wister大鼠皮下脂肪组织中分离出ASC，在含有血管内皮细胞生长因子（VEGF-A， IGF）和5% FBS的EBM或EGM培养基中培养一周。采用链脲佐菌素（STZ）注射雄性Wister大鼠。STZ给药后第9周，将上述两种方法培养的干细胞分别注射到模型大鼠和正常大鼠的阴茎海绵体中。测量平均动脉血压（MAP）和海绵体内压（ICP）。我们使用慢病毒（LV）表达大鼠AM siRNA和AdAM，研究了ASC产生的AM是否在ASC诱导的勃起功能恢复中起关键作用。结果：1；ASC管理恢复ICP；ASC可显著提高ICP/MAP和AUC。2. ASC可恢复海绵体结构；注射ASC后，海绵体小梁变大，分布更密。3. ASC生产AM；ASC在培养基中分泌AM呈时间依赖性。用表达AM siRNA的LV感染ASC，培养1周后，ASC分泌AM明显受到抑制。4. AM下调及AM强制表达的影响用表达AMsiRNA （LV_AMsiRNA）的LV或其阴性对照NC siRNA（LV_NC siRNA）感染ASC，培养1周后注射于海绵体。当ASC感染LV_NC siRNA时，通过ICP/MAP和AUC评估，ASC显著恢复勃起功能。相比之下，当ASC感染LV_AM siRNA时，ASC诱导的ICP/MAP和AUC恢复明显减弱。结论：这些结果表明，ASC具有恢复勃起功能的潜力，ASC产生的AM在恢复勃起功能中起主要作用，特别是在EGM中

INTRODUCTION AND OBJECTIVES : Diabetic patients are more refractory to the treatment with phosphodiesterase type 5 inhibitors(PDE-5I) because of severe damage in their cavernous body. We have previously reported that acute administration of Adrenomedullin(AM) into the cavernous body enhances erectile function in rats. During the screening of cytokines that adipose tissue-derived stem cells(ASC) produce, we found that ASC produced AM especially when ASC were cultured in medium containing growth factors for vascular endothelial cells(VEC). In this study, we examined whether AM would be implicated in ASCinduced restoration of erectile function.METHODS : ASC isolated from the subcutaneous adipose tissue of a Wister rat was cultured for one week in either EBM or EGM medium which contains vascular endothelial cell growth factor such as VEGF-A, IGF with 5% FBS. Streptozotocin(STZ) was injected into a male Wister rat. Stem cells cultured using the two methods above were injected into the corpu … More s spongiosum penis of the model rat and a normal rat in the ninth week after STZ administration. The mean arterial blood pressure(MAP) and the intracavernosal pressure(ICP) were measured. We used lentivirus(LV) expressing rat AM siRNA and AdAM, and examined whether AM produced by ASC played pivotal roles in ASC-induced restoration of erectile function. RESULTS : 1. ASC administration restores ICP ; Administration of ASC significantly increased ICP/MAP and AUC. 2. ASC administration restores the structure of cavernous body ; When ASC were injected, the trabeculae of cavernous body became larger and were distributed more densely. 3. ASC produce AM ; The ASC secreted AM in the culture medium in a time-dependent fashion. When ASC were infected with LV that expresses AM siRNA and cultured for 1 week, AM secretion by ASC was significantly suppressed. 4. Effects of AM knockdown and forced expression of AM ; ASC were infected with LV expressing AM siRNA(LV_AMsiRNA) or its negtive control NC siRNA(LV_NC siRNA), cultured for 1 week, and injected in the cavernous body. When ASC were infected with LV_NC siRNA, ASC significantly restored erectile function, as assessed by ICP/MAP and AUC. In contrast, when ASC were infected with LV_AM siRNA, ASC-induced restoration of ICP/MAP and AUC was significantly diminished. CONCLUSIONS : These results suggested that ASC have potentials to restore erectile function and that AM produced by ASC when cultured especially in EGM plays a major role in the restoration of erectile function Less