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目的探讨勿动蛋白A(Nogo-A)及勿动蛋白受体(NgR)在正常墨西哥钝口螈及其中脑胆碱能神经元损伤后脑组织中的表达。方法以正常墨西哥钝口螈脑组织和中脑胆碱能神经元损伤后第3、7、14、21天脑组织作为研究对象,采用免疫荧光技术观察正常墨西哥钝口螈脑组织中Nogo-A及NgR的表达;分别用荧光定量PCR和蛋白质印迹法(WB法)检测正常墨西哥钝口螈和损伤组脑组织中Nogo-A及NgR的mRNA含量和蛋白含量的变化。结果正常墨西哥钝口螈脑组织中存在Nogo-A及NgR的表达,在墨西哥钝口螈中脑胆碱能神经元损伤后(注入AF64A)第3天,中脑乙酰胆碱转移酶(ChAT)阳性细胞数明显减少,与正常组比较差异有统计学意义(P<0.01),损伤后第7、14、21天均有ChAT、5-溴脱氧尿嘧啶核苷(BrdU)双阳性细胞,且随时间的延长而增加。中脑胆碱能神经元损伤后第3、7、14、21天脑组织中,Nogo-A的mRNA和蛋白水平的表达量与正常组比较差异均无统计学意义;而NgR的mRNA和蛋白水平的表达量均下降,与正常组比较差异有统计学意义(P<0.05)。结论中枢神经系统损伤后的再生可能与NgR表达量的下降有关。
Objective To investigate the expression of Nogo-A and NgR in brain tissue of normal duodenal and injured cholinergic neuron in normal rats. Methods The brain tissues of 3 days, 7 days, 14 days and 21 days after the injury of brain and midbrain cholinergic neurons in normal Mexican Myzotis were studied. Immunofluorescence staining was used to observe the changes of Nogo-A (Nogo-A and NgR) were detected by real-time quantitative PCR and Western blot respectively. The mRNA and protein levels of Nogo-A and NgR in the brain of normal salivary gland and injury group were detected by real-time PCR and Western blot respectively. Results The expression of Nogo-A and NgR were found in the normal brain of C. mexico. Normal brain acetylcholinesterase (ChAT) positive cells were detected on the 3rd day after injection of AF64A (P <0.01). ChAT and BrdU double positive cells were detected on the 7th, 14th and 21st day after injury, respectively, The increase of the extension. There was no significant difference in the mRNA and protein expression levels of Nogo-A at 3, 7, 14, and 21 days after injury in the midbrain cholinergic neurons compared with the normal group. However, the mRNA and protein levels of NgR The level of expression decreased, compared with the normal group, the difference was statistically significant (P <0.05). Conclusion The regeneration of central nervous system after injury may be related to the decrease of NgR expression.