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N^ε-羧甲基赖氨酸通过RAGE促进平滑肌细胞源性泡沫细胞形成 被引量:1

CML/RAGE signal induces vascular smooth muscle cell derived-foam cells formation
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摘要 目的研究Nε-羧甲基赖氨酸(CML)对平滑肌细胞泡沫化中脂质外流的影响,探讨其在促进平滑肌细胞泡沫化中的作用和机制。方法使用组织贴块法从C57BL/6J小鼠主动脉中提取原代血管平滑肌细胞并进行鉴定;将第3~9代的原代平滑肌细胞分为对照组,氧化型低密度脂蛋白(ox-LDL)(50 mg/L)模型组,1、10、100μmol/L的CML与ox-LDL共刺激组,通过NBD-胆固醇流出实验检测CML对泡沫细胞胆固醇流出率的影响,通过胆固醇含量测定试剂盒测定细胞内总胆固醇(TC)、游离胆固醇(FC)和胆固醇酯(CE)水平;将原代血管平滑肌细胞分为四组:对照组、ox-LDL模型组、ox-LDL+CML组、RAGE siRNA沉默组(ox-LDL+CML+siRAGE组)并进行刺激,通过Western blot检测糖基化终末产物受体(RAGE)和胆固醇流出调节子ABCA1的表达;通过油红O染色和油红萃取实验检测各组平滑肌细胞泡沫化程度。结果与对照组相比,不同浓度CML明显升高平滑肌细胞内TC、CE和FC的含量;胆固醇流出实验表明胆固醇流出率随CML浓度升高而降低;Western blot实验表明与对照组相比,加入CML后RAGE明显升高,ABCA1明显降低,ox-LDL+CML+siRAGE组RAGE的表达降低,并且ABCA1降低水平减少;油红O染色显示ox-LDL能够促使平滑肌细胞内脂质蓄积,CML能够增强其作用,抑制RAGE表达后CML作用减弱。结论 CML通过RAGE抑制脂质外流,促进平滑肌细胞源性泡沫细胞形成。 Aim To study the effect and mechanism of Nε-carboxymethyllysine( CML) on vascular smooth muscle cells( VSMC) foam cells formation. Methods Primary VSMC were extracted from the aorta of C57 BL/6 J mice using tissue affixing method and identified; Primary smooth muscle cells of 3 ~ 9 passages were divided into control group,oxidized low-density lipoprotein( ox-LDL)( 50 mg/L) group,1,10,and 100 μmol/L concentrations of CML and ox-LDL co-stimulation group. The levels of total cholesterol( TC),free cholesterol( FC) and cholesteryl esters( CE) were determined by a cholesterol content assay kit; the primary VSMC were divided into four groups: control group,ox-LDL-induced model group,and ox-LDL co-stimulated with CML group,RAGE siRNA silencing group. Western blot and qRT-PCR was used to detect receptor for advanced glycation end products( RAGE) and cholesterol efflux end regulator ATP binding cassette transporter A1( ABCA1); The degree of foaming of smooth muscle cells in each group was detected by oil red O staining and oil red extraction experiments. Results Compared with control group,CML significantly increased smooth muscle intracellular TC,CE and FC levels; Cholesterol efflux experiments showed that cholesterol efflux rate decreased withincreasing CML concentration; Western blot showed that compared with the control group,RAGE in the CML group was significantly higher,ABCA1 was significantly lower,and RAGE siRNA decreased expression of RAGE,and increased ABCA1 levels. Oil red staining showed that ox-LDL could promote lipid accumulation in smooth muscle cells,and CML could enhance its effect. Conclusion CML/RAGE inhibits VSMC cholesterol efflux,induces vascular smooth muscle cell derived-foam cells formation.
作者 包正阳 孙振 李丽华 王小东 宋娟 邵晨 严金川 王中群 BAO Zhengyang1,SUN Zhen1,LI Lihua2,WANG Xiaodong1,SONG Juan1,SHAO Chen1;YAN Jinchuan1;WANG Zhongqun1 ( 1.Department of Cardiology, 2. Department of Pathology, Affiliated Hospital of Jiangsu University, Zhenjiang, Jiangsu 212001, China)
出处 《中国动脉硬化杂志》 2018年第7期672-677,共6页 Chinese Journal of Arteriosclerosis
基金 国家自然科学基金(81770450,81370408) 江苏省研究生科研创新计划(KYCX17_1801) 镇江市社会发展项目(SH2016037,SH2017031)
关键词 N^ε-羧甲基赖氨酸 晚期糖基化终末产物受体 胆固醇外流 血管平滑肌细胞 泡沫细胞形成 Nε-carboxymethyllysine receptor for advanced glycation end products cholesterol efflux vascular smooth muscle cells foam cells formation
作者简介 包正阳,硕士研究生,主要研究方向为动脉粥样硬化的基础与临床,E-mail为txwdbzy9307@outlook.com。;通信作者 王中群,博士,副研究员,博士研究生导师,主要研究方向为动脉粥样硬化基础与l艋床,E-mail为wangtsmc@aliyun.com。
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