尼曼-匹克C1型类似蛋白1介导依泽替米贝对RAW264.7细胞脂质蓄积的抑制作用  

NPC1L1 contributes to the inhibition of lipid accumulation induced by ezetimibe in RAW264.7 cells

作　　者：李靓[1] 袁皓愉 唐振旺[3] 于伟霞[1] 谢志忠[1] 庹勤慧[1] 廖端芳[1] 

机构地区：[1]南华大学药学与生命科学学院,湖南衡阳421001 [2]株洲市芦淞区疾病预防控制中心,湖南株洲412000 [3]南华大学附属第一医院,湖南衡阳421001

出　　处：《中国药理学通报》2009年第12期1563-1566,共4页Chinese Pharmacological Bulletin

基　　金：国家重点基础研究发展计划(973计划)资助项目(No2006CB503808);国家自然科学基金资助项目(No30770868)

摘　　要：目的观察肠道胆固醇吸收抑制剂依泽替米贝(ezetimibe)对RAW264.7细胞源性荷脂细胞脂质蓄积的影响并对其机制进行初步探讨。方法采用油红O染色、高效液相色谱法检测细胞内脂滴数量和细胞内脂质含量,Western blot对NPC1L1(Niemann-Pick type C1Like-1)进行定性和半定量检测。结果RAW264.7细胞中有NPC1L1蛋白表达。不同浓度(0、0.003、0.01和0.03mol.L-1)依泽替米贝预先孵育RAW264.7细胞24h或最佳浓度(0.03mol.L-1)预先孵育不同时间(0、6、12和24h)后,换50mg.L-1oxLDL继续孵育24h,结果显示不同浓度ezetimibe预先孵育后,细胞内脂滴数量与面积随着浓度的增加而逐渐减少;Ezetimibe预先孵育可减少细胞内脂质蓄积,并呈浓度和时间依赖性。其中0.03mol.L-1ezetimibe预先孵育24h组作用最明显,CE百分比较oxLDL单独孵育组减少了约47%±0.1%。结论小鼠源性巨噬细胞RAW264.7中存在NPC1L1蛋白表达;依泽替米贝能够减少RAW264.7细胞中NPC1L1蛋白表达;依泽替米贝抑制RAW264.7细胞中脂质蓄积。Aim To investigate the effect of intestinal cholesterol absorption inhibitor Ezetimibe on lipid accumulation in RAW264.7 cells and identify the underlying mechanism.Method RAW264.7 cells were pretreated with the indicated concentrations of Ezetimibe （0,0.003,0.01 and 0.03 mol·L^-1for 24 hours or pretreated with the optimal concentration（0.03 mol·L^-1of Ezetimibe for different periods （0,6,12 and 24 h）,followed by incubation with 50 mg·L-1 oxLDL for 24 hours,then the number of intracellular lipid droplets and lipid content were measured by using oil red O staining and HPLC;the expression of NPC1L1 was measured by Western blot.Results Pretreatment with indicated concentrations of Ezetimibe caused a concentration-dependent inhibition of intracellular lipid accumulation;pretreatment with 0.03 mol·L^-1Ezetimibe caused a time-dependent inhibition of intracellular lipid accumulation.It was noted that pretreatment with 0.03 mol·L^-1Ezetimibe for 24 hours inhibited CE by about 47%＋0.1% compared with control group（oxLDL alone）.Immunoblotting results showed that NPC1L1 was expressed in RAW264.7 cells and it was down-regulated after Ezetimibe treatment.Conclusions Ezetimibe causes concentration-dependent and time-dependent inhibition of lipid accumulation in RAW264.7 cells;it also reduces NPC1L1 expression in RAW264.7 cells.

关 键 词：药理学 依泽替米贝 巨噬细胞 脂质蓄积 尼曼-匹克C1型类似蛋白1 抑制作用 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R344.81[医药卫生—基础医学]