辛伐他汀对oxLDL诱导的巨噬细胞活性氧及IL-1β分泌的影响  

Effect of simvastatin on production of reactive oxygen species and secretion of IL-1β in macrophages induced by oxLDL

作　　者：靳梦醒 闫海[1] 程艳伟[1] 桂丽[1] 胡春松[1] 张林杰[1] 黄保军[1] 

机构地区：[1]安徽医科大学基础医学院免疫学教研室,安徽合肥230032

出　　处：《中国药理学通报》2014年第5期692-696,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 30972791);安徽医科大学博士科研基金(No XJ2006005)

摘　　要：目的探讨辛伐他汀对氧化低密度脂蛋白(oxidized low density lipoprotein,oxLDL)诱导的巨噬细胞内活性氧(ROS)的水平及IL-1β分泌的影响。方法将oxLDL 0、50、100、200 mg·L-1作用于巨噬细胞J774A.1后,采用油红O染色观察脂滴在巨噬细胞内的聚集情况;用辛伐他汀0.5、1.0μmol·L-1作用于oxLDL处理的巨噬细胞,流式细胞仪检测辛伐他汀作用前后细胞内ROS的水平;Western blot检测pro-caspase-1、cleaved caspase-1和成熟IL-1β的表达情况。结果油红O染色结果表明,oxLDL可导致巨噬细胞内明显的脂质聚集,并于100 mg·L-1作用剂量达到摄取脂质的饱和状态;流式细胞仪结果表明,oxLDL可以诱导脂质聚集巨噬细胞内ROS的水平增加,辛伐他汀作用后,ROS水平由(167±0.47)%下降到(139±0.97)%,并呈明显的剂量依赖;Western blot结果表明,辛伐他汀可抑制oxLDL诱导的caspase-1活化及IL-1β的分泌,辛伐他汀处理组与oxLDL实验组相比,cleaved caspase-1及成熟IL-1β的表达呈下降趋势,灰度值差异有统计学意义(P<0.05)。结论在oxLDL诱导的巨噬细胞脂质聚集模型中,辛伐他汀可以抑制巨噬细胞内ROS的产生及caspase-1的活化,并减少IL-1β的分泌。Aim To study the effect of simvastatin on the production of reactive oxygen species （ROS） and the secretion of interleukin-1 beta （IL-1β） in oxidized low density lipoprotein （oxLDL）-induced macrophages. Methods After the murine macrophage J774A. 1 was treated with 0,50,100,200 mg . L-1 oxLDL, the contents of aggregated lipid in macrophages were observed and determined by oil red O staining. Then, the oxLDL-primed macrophages were treated with 0.5,1.0 umol . L- 1 simvastatin, the production of ROS was determined by flow cytometry and the expressions of procaspase-1, cleaved caspase-1 and mature IL-1β on protein level were determined by Western blot. Results The oil red O staining results showed that oxLDL could induce obvious lipid aggregation in macrophages, and reached the saturation point with 100 mg . L-1 concentration. Flow cytometry resuhs indicated that oxLDL could induce the production of ROS in macrophages,up to 167%±0. 47%, and ROS level decreased to 139% ±0.97% in a dose-dependent manner after treatment with simvastatin. Western blot indicated that simvastatin could inhibit the expression of cleaved caspase-1 and mature IL-1b in macrophages triggered by oxLDL; compared with oxLDL group, the expression of cleaved caspase-1 and mature IL-1β decreased in simvastatin treated group, and all results had statistical significance （P 〈 0. 05）. Conclusion In the lipid aggregation model of macrophages induced by oxLDL, simvastatin can inhibit the production of ROS, caspase- 1 activation, and secretion of IL-1β in macrophages.

关 键 词：辛伐他汀 巨噬细胞 活性氧 IL-1Β caspase-1 OX-LDL 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R392.12[医药卫生—基础医学]