Wihadmadyatami Hevi, Zulfikar Muhammad Ali, Herawati Herawati, Karnati Srikanth, Saragih Golda Rani, Aliffia Dinda, Pratama Dyah A O A, Handayani Nurrahmi, Kustiati Ulayatul, Tirtosari Dewi Ratih, Tjahjono Yudy
Department of Anatomy, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia.
2Department of Chemistry, Faculty of Mathematics and Science, Institut Teknologi Bandung, Bandung, Indonesia.
Open Vet J. 2025 Jan;15(1):151-161. doi: 10.5455/OVJ.2025.v15.i1.14. Epub 2025 Jan 31.
Neurodegenerative diseases (NDDs) are distinguished by impairment and depletion of nerve cells; one of the most common NDDs is Alzheimer's disease (AD), which can appear in early onset or late onset. In recent years, the secretome or conditioned medium of mesenchymal stem cells has provided new hope for improving conditions and preventing AD. One of the secretomes is bovine umbilical mesenchymal stem cells-conditioned medium (BUMSC-CM), where BUMSC is predicted to promote neuronal proliferation potentially.
This study analyzes the therapeutic efficiency of conditioned medium or secretome produced from BUMSC-CM in treating neurodegeneration in animal models of AD.
Five groups consisting of 12 male rats were assigned: untreated (Group A, = 5), positive control group given normal saline 1 ml/100 g BW (Group B, = 5), AD rats model followed by Donepezil treatment (Group C, = 5), AD rats model with BUMSC-CM 0.2 ml/kg BW post-trimethyltin (TMT) induction (Group D, = 5), and AD rats model with BUMSC-CM 0.5 ml/kg BW post-TMT induction (Group E, = 5). The brain samples were analyzed for neuronal density using cresyl violet staining. The expression and activity of brain-derived neurotrophic factor (BDNF) were analyzed by ELISA; in addition, interleukin 1beta (IL-1β), tumor necrotic factor-alpha (TNF-α), and neural growth factor (NGF) were analyzed by quantitative polymerase chain reaction. Interactions between the main substances of BUMSC-CM and beta-amyloid protein were visualized using molecular docking.
Our result demonstrated that BUMSC-CM with the dosage of 0.5 ml/kg BW significantly increased BDNF concentration. We also found that BUMSC-CM with dosage 0.2 ml/kg BW and 0.5 ml/kg BW down-regulated IL-1β and TNF-α and upregulated NGF expression. Additionally, the number of neurons in AD rats post-treated with BUMSC-CM was significantly increasing. Furthermore, the amino acids in BUMSC-CM, including isoleucine, leucine, and valine, bind to the amyloid beta protein via interactions that are hydrophobic and hydrogen-bonded.
In this study, the neuroprotective potential of BUMSC-CM was demonstrated by its ability to upregulate BDNF and NGF while downregulating IL-1β and TNF-α. Additionally, BUMSC-CM showed potential to promote neuron proliferation in the hippocampus regions of a rat AD model. The main constituents in BUMSC-CM adhere to amyloid beta protein, hence diminishing the likelihood of ND disorders, specifically AD.
神经退行性疾病（NDDs）的特征是神经细胞受损和耗竭；最常见的神经退行性疾病之一是阿尔茨海默病（AD），其可早发或晚发。近年来，间充质干细胞的分泌组或条件培养基为改善病情和预防AD带来了新希望。其中一种分泌组是牛脐间充质干细胞条件培养基（BUMSC-CM），预计BUMSC可能促进神经元增殖。
本研究分析了由BUMSC-CM产生的条件培养基或分泌组在治疗AD动物模型神经退行性变中的治疗效果。
将12只雄性大鼠分为五组：未治疗组（A组，n = 5）、给予1 ml/100 g体重生理盐水的阳性对照组（B组，n = 5）、给予多奈哌齐治疗的AD大鼠模型组（C组，n = 5）、三甲基锡（TMT）诱导后给予0.2 ml/kg体重BUMSC-CM的AD大鼠模型组（D组，n = 5）、TMT诱导后给予0.5 ml/kg体重BUMSC-CM的AD大鼠模型组（E组，n = 5）。使用甲酚紫染色分析脑样本中的神经元密度。通过酶联免疫吸附测定法分析脑源性神经营养因子（BDNF）的表达和活性；此外，通过定量聚合酶链反应分析白细胞介素1β（IL-1β）、肿瘤坏死因子-α（TNF-α）和神经生长因子（NGF）。使用分子对接可视化BUMSC-CM的主要物质与β-淀粉样蛋白之间的相互作用。
我们的结果表明，剂量为0.5 ml/kg体重的BUMSC-CM显著增加了BDNF浓度。我们还发现，剂量为0.2 ml/kg体重和0.5 ml/kg体重的BUMSC-CM下调了IL-1β和TNF-α并上调了NGF表达。此外，用BUMSC-CM治疗后的AD大鼠中的神经元数量显著增加。此外，BUMSC-CM中的氨基酸，包括异亮氨酸、亮氨酸和缬氨酸，通过疏水和氢键相互作用与淀粉样β蛋白结合。
在本研究中，BUMSC-CM的神经保护潜力通过其上调BDNF和NGF同时下调IL-1β和TNF-α的能力得以证明。此外，BUMSC-CM显示出促进大鼠AD模型海马区神经元增殖的潜力。BUMSC-CM中的主要成分与淀粉样β蛋白结合，从而降低了神经退行性疾病，特别是AD的发生可能性。
