Zhang Jimei, Zhu Ling, Zhou Jianping, Yu Qunying, Yang Guangyuan, Luo Chaoli, Meng Jianguo, Xing Shan, Liu Jing, Mou Donggang, Yang Xuming
Gastroenterology Department, Chenggong Hospital, Yan'an Hospital Affiliated to Kunming Medical University, Kunming, Yunnan 650505, China.
Orthopedics Department, Chenggong Hospital, Yan'an Hospital Affiliated to Kunming Medical University, Kunming, Yunnan 650505, China.
Tissue Cell. 2025 Oct;96:102972. doi: 10.1016/j.tice.2025.102972. Epub 2025 May 9.
Bone marrow mesenchymal stem cells (BMSCs) are stem cells that reside in bone marrow and have multidirectional differentiation potential. BMSCs have been used to treat bone injury. However, long-term passage leads to the aging of BMSCs and the weakening of osteogenic differentiation. Furthermore, brain-derived neurotrophic factor (BDNF) may enhance the antiaging ability of BMSCs. The purpose of this study was to investigate the role of BDNF in the senescence and osteogenic differentiation of human BMSCs (hBMSCs).
The senescence of hBMSCs was induced by successive passages. The mRNA and protein expression levels were measured using RTqPCR and Western blotting. Alkaline phosphatase (ALP) and alizarin red S (ARS) staining were used to identify osteogenic differentiation in the cells.
After long-term passage, the hBMSCs morphologically gradually expanded and appeared flat, cell viability decreased, the number of fibroblast-like colony-forming units (CFU-Fs) decreased, and the number of β-galactosidase (SA-β-gal)-positive cells and the mRNA expression levels of the senescence-related genes p53, p21 and p16 increased. The activity of ALP, the level of calcium salt deposition and the protein levels of runt-related transcription factor 2 (RUNX2), osteocalcin (OCN), osteopontin (OPN) and BDNF were significantly decreased. Subsequent research indicated that the senescence and inhibition of the osteogenic differentiation of hBMSCs induced by long-term culture were caused by low expression of BDNF. From a mechanistic standpoint, BDNF can activate the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) pathway by upregulating the expression of tropomyosin receptor kinase B (TrkB), thereby improving the senescence and inhibition of the osteogenic differentiation of hBMSCs caused by long-term passage.
BDNF improves the senescence and inhibition of the osteogenic differentiation of hBMSCs caused by long-term passage via regulation of the TrkB/PI3K/AKT signaling axis.
骨髓间充质干细胞（BMSCs）是存在于骨髓中的干细胞，具有多向分化潜能。BMSCs已被用于治疗骨损伤。然而，长期传代导致BMSCs衰老和成骨分化能力减弱。此外，脑源性神经营养因子（BDNF）可能增强BMSCs的抗衰老能力。本研究的目的是探讨BDNF在人BMSCs（hBMSCs）衰老和成骨分化中的作用。
通过连续传代诱导hBMSCs衰老。采用RTqPCR和蛋白质免疫印迹法检测mRNA和蛋白质表达水平。用碱性磷酸酶（ALP）和茜素红S（ARS）染色鉴定细胞的成骨分化。
长期传代后，hBMSCs形态逐渐变大且变扁平，细胞活力下降，成纤维细胞样集落形成单位（CFU-Fs）数量减少，β-半乳糖苷酶（SA-β-gal）阳性细胞数量以及衰老相关基因p53、p21和p16的mRNA表达水平增加。ALP活性、钙盐沉积水平以及 runt相关转录因子2（RUNX2）、骨钙素（OCN）、骨桥蛋白（OPN）和BDNF的蛋白质水平显著降低。后续研究表明，长期培养诱导的hBMSCs衰老和成骨分化抑制是由BDNF低表达引起的。从机制角度来看，BDNF可通过上调原肌球蛋白受体激酶B（TrkB）的表达激活磷脂酰肌醇3激酶/蛋白激酶B（PI3K/AKT）通路，从而改善长期传代导致的hBMSCs衰老和成骨分化抑制。
BDNF通过调节TrkB/PI3K/AKT信号轴改善长期传代导致的hBMSCs衰老和成骨分化抑制。
