论文部分内容阅读
目的探讨施万细胞对丙烯酰胺诱导的周围神经损伤的影响机制。方法(1)丙烯酰胺周围神经损伤/修复动物模型:免疫组化法检测大鼠神经元钙通道的特异位点(Synapsin-I)和施万细胞特异标志蛋白(S-100β)的变化。(2)细胞模型(原代施万细胞、背根神经节神经元单独培养+插入式培养皿介导混合培养):噻唑蓝(MTT)比色法检测不同浓度丙烯酰胺对神经细胞的损伤作用并探讨施万细胞的整体保护效应;Fura-2/AM负载荧光分光光度法测定单独和混合培养条件下神经细胞内Ca2+浓度的变化。结果丙烯酰胺染毒大鼠表现明显的周围神经毒性症状,恢复4～5周症状缓解。染毒末S-100β的阳性信号明显减弱提示施万细胞损伤,随恢复期延长,S-100β阳性信号渐增强提示施万细胞损伤后的恢复性变,Synapsin-I与S-100β的变化趋势基本一致。丙烯酰胺作用后混合培养组神经细胞的存活率(51.08%±3.24%)大于单独培养组(42.08%±5.42%),混合培养的神经细胞内钙离子浓度显著低于单独培养组。结论施万细胞影响丙烯酰胺诱导的周围神经损伤后的修复过程,钙通道是其发挥该有益作用的途径之一,SynapsinI是其关键的作用位点之一。
Objective To investigate the mechanism of Schwann cells affecting acrylamide induced peripheral nerve injury. Methods (1) Acrylamide peripheral nerve injury / repair animal model: The changes of synapsin-I and S-100β in rat neurons were detected by immunohistochemistry. (2) Cell model (Primary Schwann cells and dorsal root ganglion neurons were cultured separately and mixed with Petri dishes): MTT assay was used to detect the damage of nerve cells with different concentrations of acrylamide And to explore the overall protective effect of Schwann cells; Fura-2 / AM load spectrofluorimetric assay alone and mixed culture under the conditions of changes in intracellular Ca2 + concentration. Results Acrylamide in rats showed obvious symptoms of peripheral neurotoxicity, recovery 4 to 5 weeks to relieve symptoms. The positive signal of S-100β at the end of the exposure was significantly weakened, which indicated the injury of Schwann cells. With the prolongation of the recovery period, the positive signal of S-100β gradually increased, which indicated the recovery of Schwann cells after injury, the change tendency of Synapsin-I and S-100β Basically the same. The survival rate of neuronal cells in mixed culture group (51.08% ± 3.24%) was higher than that in cultured group (42.08% ± 5.42%) after mixed with acrylamide. The intracellular calcium concentration of mixed culture group was significantly lower than that of cultured group alone. Conclusion Schwann cells affect the repair process of acrylamide induced peripheral nerve injury. Calcium channel is one of the ways to play its beneficial role. Synapsin I is one of the key sites.