NR2B反义寡核苷酸对大鼠海马CA1区NR2B蛋白质表达的影响

目的 观察NMDA受体2B亚单位（NR2B）反义寡核苷酸（ANR2B）对海马CA1区NR2B蛋白质表达的影响，筛选有效的反义寡核苷酸，为探讨针对NR2B的新药物提供形态学基础。方法 设计、筛选、合成ANR2B。正常SD大鼠，海马CA1区立体定位注射ANR2B，灌注取脑，连续冰冻切片，ABC免疫组织化学方法染色，光镜下观察NR2B蛋白质的表达变化。结果 注射ANR2B后，注射区及其周围NR2B免疫组织化学染色强度明显下降。仅有少量锥体细胞和颗粒细胞散在分布；而在注射NR2B正义寡核苷酸（SNR2B）、生理盐水及生理盐水插针不注射的海马切片上，海马CA1区的染色特征与注射ANR2B者有明显差别，其作用区锥体细胞、颗粒细胞及顶树突的NR2B免疫组织化学染色强度无明显变化。结论 ANR2B能够降低NR2B蛋白质在正常大鼠海马CA1区的基础性高表达。

The effect of antisense oligonucleotide on the expression of NR2B in CA1 subfield of rat hippocampus

Objective To probe into the effect of antisense oligonucleotide on NMDA receptor 2B(ANR2B) in CA1 subfield of normal rat hippocampus as a basic approach to the new drug regulating the expression of NR2B protein.Methods ANR2B and sense oligonucleotide to NMDA receptor 2B(SNR2B) were prepared and injected into the normal rat hippocampal CA1 subfield under well controlled conditions.Serial cryosections of the brain were processed by ABC immunohistochemistry to examine the NR2B expression.Results The immunoreactivity of NR2B expression was obviously decreased in CA1 subfield after ANR2B injection(P<0.05), leaving only a few pyramidal and granular cells faintly stained.But the intensity of immunoreactivity was not altered at all in the control cases of normal tissue,normal saline injection,shame injection and SNR2B injection.Conclusion It is likely that the high basal expression of NR2B in the normal rat hippocampal CA1 subfield can be downregulated by using the antisense oligonucleotide technique.