Hu Daoyuan, Ge Yunlong, Xi Yuhang, Chen Jialiang, Wang Hua, Zhang Chi, Cui Yubin, He Lizhao, Su Ying, Chen Jun, Hu Cheng, Xiao Hengjun
Department of Urology, The Third Affiliated Hospital, Sun Yat-Sen University, Guangzhou, China.
Department of Urology, The Sixth Affiliated Hospital, Sun Yat-Sen University, Guangzhou, China.
World J Mens Health. 2024 Jul;42(3):638-649. doi: 10.5534/wjmh.230149. Epub 2024 Jan 2.
The poor retention and ambiguous differentiation of stem cells (SCs) within corpus cavernosum (CC) limit the cell application in erectile dysfunction (ED). Herein, the effects and mechanism of gene modification on modulating the traits and fate of bone marrow-derived mesenchymal stem cells (BMSCs) were investigated.
The effects of miR-145 on cell apoptosis, proliferation, migration, and differentiation were determined by flow cytometry, cell counting kit-8, transwell assays and myogenic induction. Then, the age-related ED rats were recruited to four groups including phosphate buffer saline, BMSC, vector-BMSC, overexpressed-miR-145-BMSC groups. After cell transplantation, the CC were harvested and prepared to demonstrate the retention and differentiation of BMSCs by immunofluorescent staining. Then, the target of miR-145 was verified by quantitative real-time polymerase chain reaction and immunohistochemical. After that, APTO-253, as an inducer of Krüppel-like factor 4 (KLF4), was introduced for rescue experiments in corpus cavernosum smooth muscle cells (CCSMCs) under the co-culture system.
, miR-145 inhibited the migration and apoptosis of BMSCs and promoted the differentiation of BMSCs into smooth muscle-like cells with stronger contractility. , the amount of 5-ethynyl-2'-deoxyuridine (EdU)cells within CC was significantly enhanced and maintained in the gene modified BMSC group. The EdU/CD31 co-staning was detected, however, no co-staining of EdU/α-actin was observed. Furthermore, miR-145, which secreted from the gene modified BMSCs, dampened the expression of KLF4. However, the effects of miR-145 on CCSMCs could be rescued by APTO-253.
Overall, miR-145 modification prolongs the retention of the transplanted BMSCs within the CC, and this effect might be attributed to the modulation of the miR-145/KLF4 axis. Consequently, our findings offer a promising and innovative strategy to enhance the local stem cell-based treatments.
阴茎海绵体内干细胞（SCs）的低保留率和模糊分化限制了其在勃起功能障碍（ED）中的细胞应用。在此，研究了基因修饰对调节骨髓间充质干细胞（BMSCs）特性和命运的影响及机制。
通过流式细胞术、细胞计数试剂盒-8、Transwell实验和生肌诱导来确定miR-145对细胞凋亡、增殖、迁移和分化的影响。然后，将年龄相关性ED大鼠分为四组，包括磷酸盐缓冲盐水组、BMSC组、载体-BMSC组、过表达-miR-145-BMSC组。细胞移植后，收获阴茎海绵体并通过免疫荧光染色来证明BMSCs的保留和分化。然后，通过定量实时聚合酶链反应和免疫组织化学验证miR-145的靶标。之后，在共培养系统下，引入APTO-253作为Krüppel样因子4（KLF4）的诱导剂，在阴茎海绵体平滑肌细胞（CCSMCs）中进行挽救实验。
miR-145抑制BMSCs的迁移和凋亡，并促进BMSCs分化为具有更强收缩力的平滑肌样细胞。阴茎海绵体内5-乙炔基-2'-脱氧尿苷（EdU）细胞数量在基因修饰的BMSC组中显著增加并得以维持。检测到EdU/CD31共染色，但未观察到EdU/α-肌动蛋白的共染色。此外，基因修饰的BMSCs分泌的miR-145抑制了KLF4的表达。然而，APTO-253可挽救miR-145对CCSMCs的影响。
总体而言，miR-145修饰延长了移植的BMSCs在阴茎海绵体内的保留时间，这种作用可能归因于miR-145/KLF4轴的调节。因此，我们的研究结果为增强基于局部干细胞的治疗提供了一种有前景的创新策略。
