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目的观察大鼠模拟失重后中脑导水管周围灰质(periaqueductafgray,PAG)神经元Fos蛋白表达的改变。方法采用雌性大鼠尾部悬吊法建立模拟失重模型,按体重配对原则随机将大鼠分为2组,即模拟失重14d(SW-14d)组和正常对照(Con)组;根据实验干预措施的不同,每组又分为3个亚组,即电刺激(electrical stimulation,ES)亚组,琥珀胆碱(succinylcholine,Sch)预处理(pretreatment ofSch,PS)亚组和单纯琥珀胆碱注射(Sch injection,SI)亚组,每个亚组4只大鼠。采用免疫组织化学ABC法对大鼠中脑冰冻切片进行染色,显微镜下进行观察并拍照,对Fos免疫阳性细胞进行形态与计数分析。结果两组大鼠PAG腹外侧区均观察到Fos样蛋白的表达。与正常对照组相比,模拟失重组Fos免疫阳性细胞,核着色较淡,边界较为模糊;Fos免疫阳性细胞数明显减少。不同亚组大鼠Fos免疫阳性细胞计数分析表现为:Sch预处理亚组>伤害性电刺激亚组>单纯琥珀胆碱注射亚组,正常对照组大鼠阳性细胞计数结果分别为71.06±8.96、46.94±3.38和35.04±4.62;模拟失重2周组分别为32.91±2.99,27.77±3.27和11.75±1.00。结论伤害性电刺激可诱发大鼠PAG腹外侧区神经元Fos样蛋白的表达,而琥珀胆碱可使其表达增加;模拟失重后伤害性电刺激诱发的Fos样蛋白在PAG的表达减少。
Objective To observe the changes of Fos protein expression in periaqueductafgray (PAG) neurons of midbrain after simulated weightlessness in rats. Methods The model of weightlessness was established by the tail suspension method in female rats. The rats were randomly divided into two groups according to the principle of body weight matching: the simulated weightlessness 14d (SW-14d) group and the normal control (Con) group. According to the experimental intervention In addition, each group was divided into three subgroups: electrical stimulation (ES) subgroup, succinylcholine (Sch) pretreatment of Schizophrenia (PS) subgroup and Schuman injection, SI) subgroup, 4 rats in each subgroup. Immunohistochemistry ABC method was used to stain the frozen sections of rat midbrain. The morphology and count of Fos immunopositive cells were observed under the microscope and photographed. Results Fos-like protein expression was observed in the ventrolateral region of PAG in both groups. Compared with the normal control group, Fos immunoreactive cells in the simulated weightlessness group showed a faint nuclear staining and a faint borderline. The number of Fos immunoreactive cells was significantly decreased. The counts of Fos immunoreactive cells in different subgroups were as follows: Sch pretreatment subgroup> nociceptive stimulation subgroup> succinylcholine injection subgroup, the positive control group rats positive cell count results were 71.06 ± 8.96, 46.94 ± 3.38 and 35.04 ± 4.62, respectively. The two weeks after simulated weightlessness were 32.91 ± 2.99, 27.77 ± 3.27 and 11.75 ± 1.00, respectively. Conclusions Noxious electrical stimulation can induce Fos-like protein expression in the neurons of rat’s PAG. However, succinylcholine increases the expression of Fos-like protein in PAG after simulated weightlessness.