d-609 and Spinal-Cord-Injuries

The rate of neuronal differentiation of bone marrow stromal cells (BMSCs) in vivo is very low; therefore, it is necessary to elevate the number of BMSC-derived neurons to cure neurodegenerative diseases. We previously reported that tricyclodecane-9-yl-xanthogenate (D609), an inhibitor of phosphatidylcholine-specific phospholipase C (PC-PLC), induced BMSCs to differentiate into neuron-like cells in vitro. However, the neuronal type is not clear, and it is still unknown whether these neuron-like cells possess physiological properties of functional neurons and whether they can contribute to the recovery of neuron dysfunction. To answer these questions, we investigated their characteristics by detecting neuronal function-related neurotransmitters and calcium image. The results showed that these cells exhibited functional cholinergic neurons in vitro. Transplantation of these cholinergic neuron-like cells promoted the recovery of spinal cord-injured mice, and they were more effective than BMSCs. The number of cholinergic neurons was increased after injection with BMSC-derived cholinergic neuron-like cells, indicating their high differentiation rate in vivo. Moreover, the proportion of cholinergic neurons in host cells and secretion of acetylcholine were increased, and preservation of neurofilament was also observed in the lesion of mice implanted with BMSC-derived neurons, suggesting the neuronal protection of BMSC-derived neurons. Our findings provide both a simple method to induce the differentiation of BMSCs into cholinergic neuron-like cells and a putative strategy for the therapy of spinal cord injuries.

Topics: Acetylcholine; Animals; Axons; Biomarkers; Bridged-Ring Compounds; Bromodeoxyuridine; Cell Differentiation; Choline O-Acetyltransferase; Cholinergic Neurons; Glutamate Decarboxylase; Graft Rejection; Immunohistochemistry; Male; Mesenchymal Stem Cells; Mice; Mice, Inbred C57BL; Motor Neurons; Nerve Regeneration; Neuroprotective Agents; Norbornanes; Rats; Recovery of Function; Spinal Cord Injuries; Thiocarbamates; Thiones; Transplantation, Heterologous

Ceramide is a sphingolipid signaling molecule with powerful proinflammatory and proapoptotic properties. The aim of this study was to investigate the role of altered ceramide metabolism in spinal cord injury. Spinal cord injury was induced by application of vascular clips (force of 24 g) to the dura via a four-level T5-T8 laminectomy. Spinal cord injury in mice resulted in severe trauma characterized by edema, neutrophil infiltration, production of a range of inflammatory mediators, tissue damage, and apoptosis. Fumonisin B1, tyclodecan-9-xanthogenate (D609), and (3-carbazol-9-yl-propyl)-[2-(3,4-dimethoxy-phenyl)-ethyl]-methylamine (NB6) inhibitors of, respectively, ceramide synthase, acid sphingomyelinase, and the secretory form of acid sphingomyelinase significantly reduced the degree of (i) ceramide formation, (ii) tissue injury, (iii) neutrophil infiltration, (iv) nitrotyrosine formation, (v) TNF-alpha and IL-1beta production and apoptosis (TUNEL staining and Bax and Bcl-2 expression). Significant improvement of motor function was observed in mice treated with inhibitors of the de novo (fumonisin B1) and sphingomyelin (D609, NB6) pathways. These results implicate ceramide in the pathogenesis of spinal cord injury, providing the rationale for development of candidates for its therapeutic inhibition.

Topics: Animals; Antioxidants; Apoptosis; Bridged-Ring Compounds; Carbazoles; Ceramides; Enzyme Inhibitors; Fumonisins; Immunohistochemistry; In Situ Nick-End Labeling; Interleukin-1beta; Male; Mice; Norbornanes; Oxidoreductases; Random Allocation; Sphingomyelin Phosphodiesterase; Spinal Cord Injuries; Thiocarbamates; Thiones; Tumor Necrosis Factor-alpha