| Objective:Carbon monoxide (CO) are acute poisoning the most common cause of death of asphyxiating gases,is the common occupational poison in our life. Can lead to central nervous system and cardiovascular system function of many organ-based damage.After acute poisoning symptoms disappeared,some patients will present normal or near normal for a few days or weeks,it called the false period. They present mainly the dementia symptomns,which known as the delayed encephalopathy after acute carbon monoxide poisoning,DEACMP. Its pathogenesis has not yet been clarified, The field has always been a thorny problem of clinical problems,Method of treatment has no effect, it creates the enormous burden for the society and the family.The Nogo-A protein is by the Nogo gene code protein, Nogo-A has suppresses the mature central nervous system neuron axon palingenesis, is thought at present the Inhibition center nervous system regenerates one of essential material. Nogo-A has been proved to be NI-250, in the central nervous system expression of oligodendrocyte, peripheral nervous system Schwann cells did not express.Nogo-A at the central nervous system are found in myelin inhibition of axon growth of a protein, it contains two completely independent of inhibitory activity with the structure of the domain: Located at cell's in amino-Nogo and located at cell surfa- -ce Nogo-66.Nogo-66 receptor are complex, with its combination of NgR/p75/Ling-1 role. Nogo-A has been confirmed by the three different regions of the hair angl inhibi- -t axon growth cone collapse, prevent the spread fibroblasts, which make Nogo-A as a limit of nerve regeneration and repair of axonal injury, a key factor in. In view of Nogo-A protein and its receptor system in the physiological character-istics of anatomy and pathology,But in the DEACMP patient skull phantom study white matter escapes the myelinic membrane to be prominent, therefore we extrapolated that the Nogo-A protein and acceptor system it relates closely especially with the acute CO poison with DEACMP. In this experiment we use the exogenetic CO direct processing in vitro raise little suddenly the spongiocyte, removes the oxygen deficit influence, observation little suddenly spongiocyte Nogo-A in the protein and the mRNA level expression influence, provides the new experiment clue for the research acute CO poison and the DEACMP concrete mechanism, brings the new opportunity and the hope for the clinical prevention.Ganglioside is a sialic acid containing nerve sphingolipids sugar substances, the composition of the human cell membrane composition, content in the central nervous system in particular, is rich in the highest concentration in brain gray matter is peripheral neuropathy in 8% ~ 20%, but the concentration of serum and cerebrospinal fluid in low.According to the number of sialic acid is usually divided into GM (single ganglioside sialic acid) GD (2 ganglioside sialic acid) and GT (III ganglioside sialic acid) and so on. According to the number of sugar-based GM will be divided into different GM1 (containing four glycosylation), GM2 (containing three glycosylation), GM3 (containing two glycosylation).Ganglioside has a wealth of biological functions, such as to maintain the stability of cell membrane structure; as cell markers and antigens involved in the interaction between cells and recognition; as differentiation markers involved in cell growth regulation and information transmission; as some biological activity substances receptors, affect cell function and plasma membrane proteins involved in cell adhesion and so on. A large number of experimental results show that the quality of ganglion glycoside substances can regulate neuronal membrane and the nerve function, especially for the differentiation and maturation of nerve tissue and nerve tissue injury in a clear break to promote the role of V; of which four have been single-sialic acid sugar nerve section of resin glycosides (GM1) on the neuronal protective effect significantly. GM1 ganglioside is one of the most important in the treatment of central nervous system lesions play an important role in neurogenesis, the growth and differentiation play an essential role. Exogenous GM1 may have entered the blood-brain barrier into the central nervous system and play a role in nerve cell membrane. GM1 on the central nervous system injury repair has obvious role。GM1 of CO poisoning caused by oligodendrocyte damage to whether the protective effects with the oligodendrocyte Nogo-A protein related to the present there is no report, to be in the cells in this experiment to look into the level.Methods:From 2 days newborn SD rat optic nerve, using tissue culture method, using chemically defined culture medium DMEM/F12 culture, purified oligodendrocyte. (2) to exclude the influence of hypoxia, 1% CO oligodendrocyte processing, using RT-PCR and immunohistochemical observation of cells, 6h, 24h, 48h oligodendrocyte Nogo-AmRNA and protein expression. (3) in the culture medium by adding pre-GM1, 1% CO to deal with oligodendrocyte, Nogo-AmRNA Detection and protein expression.Results:1. the beginning of tissue 24h adherent, 48h-72h nerve tissue can be seen from the edge of a small number of cells travel mainly oval and spindle, there is a long process. 7 ~ 9 days delay cells increased rapidly, about 9 ~ 10 days, cells from the tissue free out flat at the end of the dish. About 11 days to obtain the basic cells for the cell body was round or polygonal, diameter 6-10um, cell counts for each hole (6 ~ 8)×104. Cell-mediated immunity MBP-positive staining, more than 95% positive cells.2.RT-PCR detected the expression of Nogo-A mRNA: the expression of the control group began to rise 6 hours, 24 hours and reached the peak after 48 hours decreased expression; CO group was significantly higher than the expression of the same period last year; GM1 expression closer to control group group.3.cell-mediated immunity chemical detection Nogo-A protein expression: Nogo-A expression in the oligodendrocyte membrane and cytoplasm, in order to tan. Immunocytochemical staining image analysis results showed that: the control group 6 hours, 24 hours, 48 hours, both Nogo-A protein expression, but no time table to see significant changes; CO group at different time points the cumulative optical density significantly higher in the control group (P <0.05) differences statistically; GM1 intervention decreased the cumulative optical density close to the control group (P <0.05) differences statistically.Conclusion:1. Neonatal rat optic nerve oligodendrocytes in vitro technology and stability, available in sufficient quantities required for cell experiments.2. Induced by exogenous CO in vitro oligodendrocyte Nogo-AmRNA and increased protein expression.3. Induced by exogenous CO in vitro oligodendrocyte Nogo-AmRNA expression, the application of GM1 on protein Nogo-AmRNA and inhibit the expression.
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