神经钙调蛋白介导IL-1β诱导的NF-κBp65的表达及神经毒性  

Calcineurin mediates the NF-κB p65 expression and neurotoxic ity induced by interleukin-1β

作　　者：高玲[1] 康劲松[1] 候中赤[2] 潘慧[2] 祝世功[2] 

机构地区：[1]吉林大学基础医学院,吉林长春130021 [2]北京大学医学部生理与病理生理学系,北京100083

出　　处：《中国病理生理杂志》2005年第4期625-629,共5页Chinese Journal of Pathophysiology

基　　金：国家重点基础研究发展规划项目 (No .G2 0 0 0 0 5 6 90 8) ;国家自然科学基金资助项目 (No.30 370 5 5 7) ;教育部博士点基金资助项目 (No .2 0 0 2 0 0 0 10 83)

摘　　要：目的:探讨神经钙调蛋白在白细胞介素- 1β(IL - 1β)和兴奋性氨基酸(NMDA)诱导皮层神经元NF-κΒ表达及神经损伤机制的作用。方法:应用IL - 1β或NMDA诱导原代培养的胎鼠皮层神经元损伤;通过MTT法、LDH释放率测定观察细胞生存力和损伤程度;通过Westernblotting分析NF -κBp6 5的表达;通过膜联蛋白(An nexinV)和碘化丙啶(PI)免疫荧光法观察细胞凋亡或坏死程度。结果:MTT和LDH释放率测定结果表明,IL - 1β和NMDA都可分别引起神经细胞的生存力明显下降,并呈量效依赖关系。IL - 1β和NMDA在引起细胞损伤的同时都可促进NF -κΒp6 5的表达。应用神经钙调蛋白抑制剂CsA明显抑制IL - 1β引起的细胞损伤,也明显抑制IL - 1β诱导的NF -κBp6 5表达增加。但是,环孢菌素A(CsA)不能明显抑制NMDA诱导的细胞损伤(P >0 0 5 ) ,对NMDA诱导的NF -κΒp6 5的表达的抑制作用也不明显。AnnexinV和PI免疫荧光检测表明,IL - 1β诱导的神经损伤以神经元凋亡为主,而NMDA则主要引起坏死。结论:神经钙调蛋白参与介导IL - 1β诱导的NF -κΒp6 5的表达及神经细胞凋亡,但对NMDA诱导的神经元坏死则不起主要作用。说明神经钙调蛋白是细胞凋亡信号通路中重要分子。AIM: To explore the role of calcineurin i n th e expression of NF-κB and the neurotoxicity in cultured cortical neurons treate d with interleukin-1β (IL-1β) and NMDA. METHODS: The cultured rat cortical neurons were used in the expe riment, damage of neurons was induced by interleukin-1β(IL-1β) or excitator y amino acid (NMDA). The degree of neuron damage was examined with the methods o f MTT assay and LDH releasing rate assay, as well as the Annexin V and PI immuno fluorescence. The expression of NF-κB p65 on the neurons was tested by the West ern blot analysis. RESULTS: Viability of neurons was obviously lower in the IL-1β group and NMDA group respectively than that in control group (P<0.05). The higher expression of NF-κB p65 was induced by IL-1β or NMDA, which were coinc ident with the damage of neurons. The calcineurin inhibitor cyclosporine A (CsA) inhibited the higher expression of NF-κB p65 and neuron damage induced by IL-1 β (P<0.05) but not the NMDA (P>0.05). Annexin V and PI immunofluoresc ence showed that IL-1β mainly induced the neuron apoptosis, and NMDA induced th e neuron necrosis. CONCLUSIONS: The calcineurin mediates the higher expression of N F -κB p65 and neuron damage induced by IL-1β, but not play a critical role in th e necrosis induced by NMDA in the cultured cortical neurons. These results indic ate that calcineurin is the key molecule in the apoptotic signaling pathway.

关 键 词：白细胞介素1 钙神经素 神经元 细胞凋亡 NF—κB 

分 类 号：R363[医药卫生—病理学]