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目的探讨孕期应激对子代雌鼠成年后钙结合蛋白阳性神经元表达的影响。方法 6只SD孕鼠随机分为孕期应激组(PS组)和对照组(CON组)。PS组孕鼠在妊娠晚期接受限制性应激,CON组不给予孕期应激。子代成年后,PS组和CON组雌性子代随机分别又分为PS-S组,PS-NS组,CON-S组和CON-NS组(n均=6)。PS-S组和CON-S组动物先后接受限制性应激和冰水应激。动物在接受应激后均给与水和酒精喂养,用Western blot检测海马钙结合蛋白Parvalbumin(PV)和Calretinin(CR)蛋白的表达,进一步结合共聚焦免疫荧光组织化学方法检测PV和CR在海马CA1、CA3区和齿状回(dentate gyrus,DG)的表达。结果与CON-NS组相比,海马区PV和CR的蛋白表达在CON-S,PS-NS和PS-S组下降,以PS-S组表达最低。PV在海马CA1、CA3区和DG的免疫荧光结果与PV的蛋白表达呈一致性,免疫阳性细胞明显减少;而CR的表达除了在海马CA1区的PS-NS组略有升高外,在其余各组的表达均较减少。与CON-NS组相比,PV和CR在其它各组的蛋白表达和免疫阳性细胞计数差异有统计学意义,其中以PS-S组的表达降低最为有统计学意义(P<0.05)。结论孕期应激可影响子代雌鼠成年后海马结构钙结合蛋白PV和CR的表达。
Objective To investigate the effects of gestational stress on the expression of calcium-binding protein-positive neurons in offspring of offspring. Methods Six pregnant SD rats were randomly divided into stress group (PS group) and control group (CON group) during pregnancy. Rats in PS group received restrictive stress in the third trimester of pregnancy, and CON group did not receive intrauterine stress. After passage, the female offspring of PS and CON groups were randomly divided into PS-S group, PS-NS group, CON-S group and CON-NS group (n = 6). Animals in the PS-S and CON-S groups received restrictive stress and ice-water stress. The animals were fed with water and alcohol after receiving the stress. The expression of Parvalbumin (PV) and Calretinin (CR) protein in hippocampus were detected by Western blot. The expressions of Calretinin (CR) protein and protein in hippocampus were detected by confocal immunofluorescence histochemistry CA1, CA3 region and dentate gyrus (dentate gyrus, DG) expression. Results Compared with CON-NS group, the protein expression of PV and CR in hippocampus decreased in CON-S, PS-NS and PS-S groups, and lowest in PS-S group. The immunofluorescence results of PV in hippocampal CA1, CA3 area and DG were consistent with the expression of PV protein, and the number of immunopositive cells was significantly decreased. However, the expression of CR was slightly increased in PS-NS group in hippocampus CA1 area The expression of each group were reduced. Compared with CON-NS group, there was significant difference of protein expression and immunopositive cell count between PV and CR in other groups, and the expression of PS-S group was the most significant difference (P <0.05). CONCLUSIONS: Pregnancy stress can affect the expression of calcium and protein CR in hippocampal formation in offspring of adult offspring.