Fu Housheng, Ou Zhewen, Wang Fei, Wang Weifu, Wang Zhongyao
Department of Urology, Hainan Hospital Affiliated to Hainan Medical University, Haikou City, Hainan Province, China.
Kidney Disease Center, Hainan General Hospital, Haikou City, Hainan Province, China.
Neurourol Urodyn. 2025 Jun;44(5):1211-1218. doi: 10.1002/nau.70067. Epub 2025 May 2.
High glucose damages rat Schwann cells (SCs), which is closely related to the dysfunction of mitochondria-associated endoplasmic reticulum membranes (MAMs). Therefore, the present study aimed to investigate the protective effects and mechanisms of modified bone marrow mesenchymal stem cells (BMSCs) and mitochondrial fusion protein 2 (Mfn2) modified BMSCs against SCs injury.
The Mfn2-modified BMSCs were constructed after culturing with neural-induced differentiation solution. MAP-2 (microtubule-associated protein-2, neuron marker) and GFAP (glial fibrillary acidic protein, astrocytes marker) immunofluorescence staining was used to observe changes in the differentiation potential of neural-like BMSCs. SCs (RSC96) cells cultured under high glucose conditions were cocultured with Mfn2-modified BMSCs. Changes in functional protein expression of MAMs were detected by Western Blot. Transmission electron microscopy (TEM) was used to observe the microscopic morphology of MAMs, mitochondria and endoplasmic reticulum.
The expression level of Mfn2 was significantly increased in BMSCs transfected with Mfn2. The fluorescence densities of MAP-2 and GFAP were significantly upregulated in Mfn2-BMSCs after induction by neural inducible differentiation solution. When RSC96 was incubated with high glucose and Mfn2-modified/non-modified BMSCs, the expression level of Mfn2 in RSC96 was significantly increased, while PERK, IP3R and Drp1 expressions were significantly reduced. And the Mfn2-modified BMSCs showed more significant effects comparing to Mfn2-non-modified BMSCs. The TEM showed the structural integrity of MAMs, clear structure of mitochondrial cristae and obvious and structurally intact extension of endoplasmic reticulum in Mfn2-BMSC group.
Mfn2 transfection promoted neural-like cell differentiation in BMSCs. Mfn2-modified BMSCs modulated the structural and functional homeostasis of MAMs by regulating the expression levels of MAMs functional proteins.
高糖会损伤大鼠雪旺细胞（SCs），这与线粒体相关内质网膜（MAMs）功能障碍密切相关。因此，本研究旨在探讨经修饰的骨髓间充质干细胞（BMSCs）和线粒体融合蛋白2（Mfn2）修饰的BMSCs对SCs损伤的保护作用及机制。
用神经诱导分化液培养构建Mfn2修饰的BMSCs。采用微管相关蛋白2（MAP-2，神经元标志物）和胶质纤维酸性蛋白（GFAP，星形胶质细胞标志物）免疫荧光染色观察神经样BMSCs分化潜能的变化。将高糖条件下培养的SCs（RSC96）细胞与Mfn2修饰的BMSCs共培养。通过蛋白质免疫印迹法检测MAMs功能蛋白表达的变化。采用透射电子显微镜（TEM）观察MAMs、线粒体和内质网的微观形态。
转染Mfn2的BMSCs中Mfn2表达水平显著升高。经神经诱导分化液诱导后，Mfn2-BMSCs中MAP-2和GFAP的荧光密度显著上调。当RSC96与高糖及Mfn2修饰/未修饰的BMSCs共同孵育时，RSC96中Mfn2的表达水平显著升高，而PERK、IP3R和Drp1的表达显著降低。与未修饰Mfn2的BMSCs相比，Mfn2修饰的BMSCs显示出更显著的效果。TEM显示Mfn2-BMSC组中MAMs结构完整，线粒体嵴结构清晰，内质网延伸明显且结构完整。
Mfn2转染促进了BMSCs向神经样细胞分化。Mfn2修饰的BMSCs通过调节MAMs功能蛋白的表达水平来调节MAMs的结构和功能稳态。
