【Abstract】 Objective To investigate the effect of salidroside on rat bone marrow mesenchymal stem cells (BMSCs)differentiation into the chol inergic nerve cells, so as to provide the theory basis of the combination of salidroside and stem cellsfor cl inical therapy of nervous system diseases. Methods BMSCs were isolated from 2 Wistar rats (aged 4-6 weeks, weighing 120 g), which were identified by CD34, CD45, CD90, and CD106 with flow cytometry. According to inducing method, BMSCs at passage 2 were divided into 3 groups: In groups A and B, BMSCs were induced by salidroside (20 μg/mL) and retinoic acid (5 μmol/mL) respectively for 1, 3, 6, and 9 days, in group C, BMSCs were cultured with serum-free DMEM/F12 medium as control. MTT assay was used to detect the cellular prol iferation activity. The immunofluorescence chemical technology was used to detect the expressions of nerver growth factor (NGF) and relevant marker molecule of nerve cells, including neuron-specific enolase (NSE), microtubule-associated protein 2 (MAP2), β-Tubulin III, gl ial fibrillary acidic protein (GFAP), and the marker of cholinergic neuron, such as Acetylcholine (Ach) and NGF. RT-PCR was used to detect mRNA expressions of NSE, β-Tubulin III, GFAP, brain derived neurotrophic factor (BDNF), and γ-aminobutyric acid (GABA). ELISA was used to detect the levels of BDNF and NGF, and the expression level of NGF protein was analyzed by Western blot. Results The results of the flow cytometry showed that the cultured cells were CD90 and CD106 positive, and CD34 and CD45 negative, which indicated that the cells were BMSCs. The cellular proliferation activity in groups A and B were significantly higher than that in group C at 6 days and 9 days (P lt; 0.05). RT-PCR results showed that the expression level of NSE, BDNF, β-Tubulin III, GFAP mRNA were increased in groupA at 6 days; In group B, that expression level of NSE mRNA was up-regulated at 6 days, that expression level of BDNF mRNA increased at 1 days and reached the peak at 6 days, and that expression level of β-Tubulin III mRNA was up-regulated at 3 days, which was significantly higher than that at the other time points, and than that in group C (P lt; 0.01). But no GABA mRNA expression was detected in each group. Immunofluorescence chemical technology staining showed that the positive rates of NSE, MAP2, β-Tubulin III, and GFAP were significantly higher in group A than those in group C at 3 days; the positive rates of Ach were significantly higher at 3, 6, and 9 days than those at 1 day in groups A and B, and in groups A and B than in group C (P lt; 0.01); the positive rates of NGF in groups A and B were significantly higher than those in group C (P lt; 0.01). The levels of BDNF and NGF in groups A and B were significantly higher than those in group C at 1, 3, 6, and 9 days (P lt; 0.01), but no significant difference of BDNF was found between groups A and B (P gt; 0.05). The expression level of NGF protein in groups A and B were significantly higher than that in group C (P lt; 0.01). The NGF expression reached the peak at 6 days in group A and at 3 days in group B. Conclusion Sal idroside could induce rat BMSCs differentiate into chol inergic nerve cells in vitro.
The object of this experimental study was to investigate the influence of low-energy He-Ne laser on the motor nerve cells of the spinal cord. The experimental study included as follws: (1) Four rabbits were used in this experiment. The L5-6 spinal cord segment was irradiated by He-Ne laser percutaneously, the nerve velocity of the comon peroneal nerve was measured in order to determine the function of the spinal motor nerve cells when the peripheral nerve was intact. (2) The common peroneal nerve was transected on one side wothout repair, two weeks after laser irradiation, the grey mater of the spinal cord of L5-6 segment was procured for electronic microscopic examination. (3) The common peroneal nerve on the contralateral side was transected and followed by end-to-end anastomosis, and laser irradiation was done on the same spinal cord segment. Two weeks after irradiation, the nerve velocity of the common peroneal nerve and the toe expanding test were investigated. The results were: (1) the He-Ne laser can influence the spinal motor nerve cells function as expressed by latent rate when the peripherial nerve is intact. i.e. the nerve velocity is slower than mormal, and the amplitude is markedly decreared. (2) the change of the microstructure of the spinal motor nerve cells is comparatively slight in the 10 and 15 minutes groups. (3) the recovery of the nerve velocity and the toe expansion are more earlier in the 15 min. group. In short, the low-energy He-Ne laser can influence the function of the spinal motor nerve cells.
Epigenetic modifications include DNA methylation, RNA methylation, histone methylation, histone acetylation, and noncoding RNA. This article elucidates the molecular mechanisms by which histone modifications regulate key proteins associated with neurological disorders, focusing on how the dynamic balance between histone acetyltransferase (HAT) and histone deacetylase (HDAC) serves as an epigenetic regulatory hub, influencing disease progression through acetylation coding. It also summarizes how fluctuations in acetyl coenzyme A/nicotinamide adenine dinucleotide levels regulate cell death networks via HAT and the silence information regulator family, as well as multi-target therapeutic strategies combining HDAC inhibitors, iron chelators, and receptor-interacting protein kinase 1 inhibitors to achieve precise neuroprotection.