游离胆固醇经P38-MAPK信号途径激活未折叠蛋白反应诱导巨噬细胞凋亡  

p38-MAPK Signaling Pathway Is Involved in Apoptosis of Macrophages Induced by Free Cholesterol

作　　者：唐冰杉[1] 郭毅[1] 李方明[1] 姜昕[1] 王玲[1] 张艳波[1] 黄术良[1] 

机构地区：[1]暨南大学第二临床医学院神经内科,广东深圳518020

出　　处：《神经损伤与功能重建》2008年第6期374-376,共3页Neural Injury and Functional Reconstruction

基　　金：广东省卫生厅资助项目(WSTJJ20041028)

摘　　要：目的:探讨p38-MAPK信号通路对未折叠蛋白反应的影响及在游离胆固醇(FC)诱导巨噬细胞凋亡中的作用。方法:收集小鼠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p38特异性抑制剂SB203580进行干预,Annexin-V和PI双染后流式细胞仪检测细胞凋亡,Western-bolt检测磷酸化p38和CHOP表达。结果:普通培养基孵育的巨噬细胞无磷酸化p38和CHOP表达,只有少量细胞凋亡;而在促进FC聚集条件下孵育的巨噬细胞磷酸化p38和CHOP表达明显,8 h后凋亡细胞为(21.8±0.6)%;使用SB203580干预后无磷酸化p38表达,CHOP表达减少,凋亡细胞为(6.9±0.3)%。结论:FC聚集是诱导巨噬细胞凋亡的重要原因,p38通过激活未折叠反应参与这一过程,而SB203580通过抑制p38活性对FC诱导的巨噬细胞凋亡具有保护作用。Objective： To explore the effect of p38-MAPK signaling pathway on unfolded protein response （UPR） and subsequent apoptosis induced by free eholesterol（FC）. Methods： Macrophages were harvested from the peritonea of female C57BL6/J mice, then incubated in DMEM and 1% FBS medium alone, or medium containing acetyl-LDL and compound 58035 （F@loading conditions）, or medium containing aeetyl-LDL and compound 58035 plus SB203580 respectively. Western-blotting was used to determine the expression of p38 and CHOP. The apopto- sis percentages of macrophages were detected with flow cytometry （FCM） after the staining of Annexin-V and PI. Results ： Macrophages incubated in DMEM and 1% FBS alone did not show strong evidence of apoptosis and expression of phosphor-p38 and CHOP. After 8 h incubation in medium containing acetyl-LDL and compound 58035 in our cell culture model, the expression and phosphorylation of p38 increased notably and about （21.8 ± 0.6）% of the cells have been detected. But when macrophages cultured in medium containing acetyl-LDL and compound 58035 plus SB203580, there was no phosphor-p38 expression and the apoptosis percentages of macrophages were decreased markedly. The expression of CHOP also has been inhibited notably. Conclusion： Excess intracellular FC accumula- tion induced by macrophage apoptosis dependent on URP and the p38 is involved in this process. SB203580 can inhibit the activation of p38, which protects macrophage from apoptosis induced by FC loading.

关 键 词：游离胆固醇 P38-MAPK 巨噬细胞 凋亡 

分 类 号：R741[医药卫生—神经病学与精神病学] R741.02[医药卫生—临床医学]