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[目的]调查〔3H〕DFP在各脊髓段神经细胞膜上的特异结合。[方法]鸡曝露于Tri-o-cre-sylphosphate(TOCP)后,隔6、24、48h处死,取颈、胸、腰段脊髓,称重后制成匀浆。经超速离心,获得膜蛋白。在脊髓膜蛋白中加入8nM的〔3H〕DFP,或加入80nM的非标记DFP后又加入8nM的〔3H〕DFP,然后培养1h。用nitrocellulosefilter进行快速真空滤过,用Tris-HCl-NaCl液对滤纸冲洗,再加5mL的Aquasol-2后,计数〔3H〕DFP的结合量。[结果]对照组鸡的颈、胸、腰椎段脊髓神经细胞膜上的〔3H〕DFP特异结合量分别为832.0、857.0、864.0fmol/mg,TOCP曝露组为273.5、243.52、71.5fmol/mg。TOCP曝露组各段脊髓神经细胞膜上的〔3H〕DFP特异结合量显著低于对照组,而各段脊髓神经细胞膜之间〔3H〕DFP的特异结合量无显著性差异。随着TOCP曝露后的时间的推移,各段脊髓神经细胞膜上的〔3H〕DFP特异结合量逐渐增高,提示迟发性神经毒性有机磷化合物的特异结合膜蛋白是较均匀地分布在整个脊髓神经细胞膜上。在这些脊髓神经细胞膜上的特异性结合部位,TOCP和DFP之间有竞争性抑制作用。[结论]脊髓神经细胞膜上的特异性结合膜蛋白可能与有机磷化合物的迟发性神经毒性诱发有关。
[Objective] To investigate the specific binding of [3H] DFP to the membrane of neurons in each spinal cord segment. [Methods] After exposure to Tri-o-cre-sylphosphate (TOCP), the chickens were sacrificed at 6, 24 and 48 hours. The neck, thoracic and lumbar spinal cord were harvested and weighed. The membrane protein is obtained by ultracentrifugation. 8nM of [3H] DFP was added to the spinal membrane protein, or 8nM of [3H] DFP was added to 80nM of unlabeled DFP, followed by incubation for 1 hour. Rapid vacuum filtration through a nitrocellulose filter followed by rinsing the filter paper with Tris-HCl-NaCl solution followed by 5 mL of Aquasol-2 was used to count the amount of [3H] DFP binding. [Result] The specific binding amount of [3H] DFP on the neurons of cervical, thoracic and lumbar segments in control group was 832.0, 857.0 and 864.0 fmol / mg respectively, and that in TOCP exposure group was 273.5, 243.52 and 71.5 fmol / mg. The [3H] DFP specific binding of spinal cord neurons in each segment of TOCP exposure group was significantly lower than that of the control group, while there was no significant difference in the specific binding amount of [3H] DFP between the spinal cord neurons. With the passage of time after TOCP exposure, the specific binding of [3H] DFP on the spinal cord neurons gradually increased, suggesting that the specific binding membrane proteins of delayed neurotoxicity organophosphorus compounds are more evenly distributed throughout the spinal cord nerve Cell membrane. There is a competitive inhibition between TOCP and DFP at specific binding sites on these spinal cord neuronal membranes. [Conclusion] The specific binding membrane proteins on the membrane of spinal cord neurons may be related to the delayed neurotoxicity induced by organophosphorus compounds.