CAJ | 학술논문

背景：前期研究表明，海马内注射红藻氨酸海可诱发兴奋性红藻氨酸受体KA1亚受体在海马神经元的表达明显上调，内质网应激标志物磷酸化真核翻译起始因子2α表达增加并伴随细胞死亡。目的：探讨红藻氨酸海马内注射后内质网应激发生的机制。方法：取昆明小鼠32只，将0.15 nmol 红藻氨酸注入海马CA1区域，注射时间为60 s。分别于红藻氨酸注射后第1，2，3，4，5，6，8，12小时灌注取脑，灌注取脑前进行Bederson体征评分，然后行全脑切片FJB染色分析与免疫荧光双标记观察。结果与结论：①红藻氨酸注射后第3，4，5，6，8小时，Bederson 体征评分表明中枢神经功能出现明显损伤，FJB染色示小鼠海马内神经元死亡明显；注射后第1，2，12小时，Bederson体征评分中枢神经功能未见明显损伤，FJB染色小鼠海马神经元死亡结果不明显。②根据FJB结果，取第3，8小时的脑片做免疫组化。海马内注射红藻氨酸后导致海马神经元中KA1和磷酸化真核翻译起始因子2α在相同的时间点表达明显上调，将KA1与磷酸化真核翻译起始因子2α图片结果进行叠加处理，两者完全重合，表明KA1的表达和内质网应激发生在同一个神经细胞内。结果表明红藻氨酸首先诱导了兴奋性膜上受体KA1表达的上调，其KA1的表达上调可能引起细胞内质网功能紊乱，导致内质网应激反应，并进一步促进了神经细胞的死亡。
배경：전기연구표명，해마내주사홍조안산해가유발흥강성홍조안산수체KA1아수체재해마신경원적표체명현상조，내질망응격표지물린산화진핵번역기시인자2α표체증가병반수세포사망。목적：탐토홍조안산해마내주사후내질망응격발생적궤제。방법：취곤명소서32지，장0.15 nmol 홍조안산주입해마CA1구역，주사시간위60 s。분별우홍조안산주사후제1，2，3，4，5，6，8，12소시관주취뇌，관주취뇌전진행Bederson체정평분，연후행전뇌절편FJB염색분석여면역형광쌍표기관찰。결과여결론：①홍조안산주사후제3，4，5，6，8소시，Bederson 체정평분표명중추신경공능출현명현손상，FJB염색시소서해마내신경원사망명현；주사후제1，2，12소시，Bederson체정평분중추신경공능미견명현손상，FJB염색소서해마신경원사망결과불명현。②근거FJB결과，취제3，8소시적뇌편주면역조화。해마내주사홍조안산후도치해마신경원중KA1화린산화진핵번역기시인자2α재상동적시간점표체명현상조，장KA1여린산화진핵번역기시인자2α도편결과진행첩가처리，량자완전중합，표명KA1적표체화내질망응격발생재동일개신경세포내。결과표명홍조안산수선유도료흥강성막상수체KA1표체적상조，기KA1적표체상조가능인기세포내질망공능문란，도치내질망응격반응，병진일보촉진료신경세포적사망。
BACKGROUND:Previous studies have shown that kainic acid injected into hippocampus can significantly upregulate the expression of excitatory KA1 subunit of the kainate receptor in the hippocampus, and endoplasmic reticulum stress markers, phosphorylation of the alpha subunit of eukaryotic initiation factor 2, accompanied by celldeath. OBJECTIVE:To explore the mechanism of endoplasmic reticulum stress after kainic acid is injected into the hippocampus.METHODS:0.15 nmol kainic acid was injected into the hippocampal CA1 region of 32 adult male Kunming mice, the injection time was 60 seconds. At different time points (1, 2, 3, 4, 5, 6, 8 and 12 hours) after kainic acid was injected, the Bederson score analysis was performed, and then the brain was harvested after cerebral perfusion. FJB staining of brain sections and immunofluorescence double labeled observation were also performed. RESULTS AND CONCLUSION:(1) At 3, 4, 5, 6, 8 hours after kainic acid injection, Bederson score showed severe injury of central nervous system function, and FJB staining showed the increased of celldeath in the hippocampus (P<0.05);At 1, 2, 12 hours after injection, Bederson score showed no obvious injury of central nervous system function, and FJB staining showed unobvious celldeath in the hippocampus (P>0.05). (2) According to the results of FJB staining, the brain sections were selected at 3, 8 hours for immunohistochemistry. The expressionlevels of KA1 receptors and endoplasmic reticulum stress marker P-eIF2αwere up-regulated at the same time after kainic acid was injected into hippocampus. Two single-staining KA1 and P-eIF2αimmunofluorescence images were synthesized into one over-lapped double-stained image, and two images overlapped, indicating that the up-regulated expression of KA1 and endoplasmic reticulum stress occurred in the same nerve cells. Kainic acid first up-regulated the excitatory receptor KA1 expression, which may cause cellendoplasmic reticulum dysfunction and result in the endoplasmic reticulum stress response, further promoting neuronal celldeath.