JNK信号通路在游离胆固醇诱导巨噬细胞凋亡中的作用  

Effects of JNK signaling pathway on free cholesterol induced macrophages apoptosis

作　　者：李方明[1] 郭毅 孙圣刚[1] 唐冰杉 姜昕 李劲频[1] 王启章 王玲 

机构地区：[1]华中科技大学同济医学院附属协和医院神经内科,武汉博士研究生430022 [2]暨南大学第二临床医学院(深圳市人民医院)神经内科

出　　处：《卒中与神经疾病》2008年第2期106-109,共4页Stroke and Nervous Diseases

基　　金：广东省卫生厅资助项目(编号WSTJJ20041028)

摘　　要：目的研究JNK信号通路在游离胆固醇诱导巨噬细胞凋亡中的作用。方法收集兔腹腔巨噬细胞进行体外培养,使用100μg/ml乙酰低密度脂蛋白及10μg/ml胆固醇乙酰转移酶抑制剂-58035促进FC聚集,以JNK特异性抑制剂SP600125进行干预,Annexin-V和PI双染后用流式细胞仪检测细胞凋亡,用Western-bolt检测JNK蛋白表达。结果普通培养基孵育的巨噬细胞无磷酸化JNK表达,只有少量细胞凋亡;普通培养基加上胆固醇乙酰转移酶抑制剂和乙酰低密度脂蛋白孵育的巨噬细胞磷酸化JNK表达明显,8h后凋亡细胞占总细胞的(19.8±0.6)%;使用JNK特异性抑制剂SP600125可以有效抑制JNK活性,减少细胞凋亡。结论游离胆固醇聚集是诱导巨噬细胞凋亡的重要原因,JNK信号途径参与这一过程,JNK特异性抑制剂SP600125通过抑制JNK活性对游离胆固醇诱导的巨噬细胞凋亡具有保护作用。Objective To explore the effect of the JNK signaling pathway on free cholesterol（FC） induced macrophages apoptosis. Methods Macrophages were harvested from the peritonea of New Zealand White rabbit, then incubated in DMEM and 1% （w/v） FBS medium alone, or medium containing acetyl-LDL and compound 58035 （FC-loading conditions）, or medium containing acetyl-LDL and compound 58035 plus SP600125 respectively. Western-blot was used to determine expression of JNK. The apoptosis percentages of macrophages were detected with flow cytometry （FCM） after the staining of Annexin-V and PI. Results Macrophages incubated in DMEM and 1% （w/v） FBS alone no expression of phosphor-JNK and showed very little evidence of cell apoptosis. After 8 h of incubated with medium containing acetyl-LDL and compound 58035 （FC-loading conditions） in our cell culture model, The expression and phosphorylation of c-Jun increased notable and about （19. 8 ± 0. 6）% of the cells have been detected. But when macrophages cultured in medium con- taining acetyl-LDL and compound 58035 plus SP600125, there in no phosphorylation of c-Jun expression and the apoptosis percentages of macrophages were decreased markedly. Conclusions These results indicate that excess intracellular FC accumulation is an important reason for macrophage apoptosis and the JNK signaling pathway is involved in this process. SP600125 inhibits the activation of JNK, and therefore protects macrophage from apoptosis induced by FC loading.

关 键 词：游离胆固醇 JNK 巨噬细胞 凋亡 

分 类 号：R743[医药卫生—神经病学与精神病学]