摘要
目的通过观察泡沫细胞形成、脂滴形态以及脂滴表面蛋白分子表达的情况,探讨血清淀粉样P物质（SAP）对泡沫细胞发生发展的影响。方法利用THP-1细胞,通过佛波酯（PMA）处理48 h,诱导其分化成为巨噬细胞,再用氧化型低密度脂蛋白（ox-LDL）联合脂多糖（LPS）诱导其成为泡沫细胞作为对照组,实验组在上述过程中加入不同浓度SAP（10、20 mg/L）进行干预。采用油红O染色分析SAP对泡沫细胞形成的影响,观察脂质蓄积;同时利用Bodipy493/503染色,在荧光显微镜下观察泡沫细胞内脂滴形态及数量;提取上述泡沫细胞内的脂质,通过试剂盒定量测定细胞内甘油三酯（TG）和胆固醇酯（CE）含量;利用Western blot对脂滴表面相关的蛋白进行分析,如乙酰辅酶A羧化酶（ACC1）、脂肪分化相关蛋白（ADRP）、47 k Da的尾连蛋白（TIP47）、组蛋白去甲基化酶（JMJD3）、脂肪酸合成酶（FAS）的表达水平。结果油红O染色和Bodipy493/503染色显示,实验组细胞内的脂滴大小更大、数量更多,且20 mg/L SAP干预组较10 mg/L SAP干预组趋势更明显;脂质定量检测结果显示,实验组TG、CE含量增高,20 mg/L SAP干预组较10 mg/L SAP干预组更高; Western blot结果显示,SAP能够增强ADRP、TIP47、JMJD3、FAS的表达水平,除了FAS在不同SAP浓度干预组表达水平一样外,其余均随着SAP浓度的增加表达水平也增强。结论 SAP能够促进泡沫细胞形成,增强脂质蓄积; SAP能够增强相关脂蛋白的表达水平。
Aim To investigate the effect of serum amyloid P component（SAP） on the formation and development of foam cells by observing the formation of foam cells,the morphology of lipid droplets and the expression of lipid droplet surface protein molecules. Methods THP-1 cells were induced to differentiate into macrophages after 48 hours of PMA treatment,and then foam cells were induced by oxidized low density lipoprotein（ox-LDL） combined with lipopolysaccharide（LPS） as control group. Different concentrations of SAP（10,20 mg/L） were added to the experimental group for intervention. Oil red O staining was used to analyze the effect of SAP on foam cell formation and lipid accumulation,and Bodipy493/503 staining was used to observe the morphology and quantity of lipid droplets in foam cells under fluorescence microscope. The lipids in the foam cells were extracted and the contents of triglyceride（TG） and cholesterol ester（CE） in the foam cells were quantitatively determined by kit. Western blot was used to analyze lipid droplet surface related proteins,such as acetyl coenzyme A carboxylase（ACC1）,adipogenesis related protein（ADRP）,47 k Da telopenin（TIP47）,histone demethylase（JMJD3）,fatty acid synthase（FAS）. Results Oil red O staining and Bodipy493/503 staining showed that intracellular lipid droplets in the experimental group were larger and more numerous,and the trend was more obvious in the 20 mg/L SAP intervention group than in the 10 mg/L SAP intervention group. The results showed that SAP could enhance the expression levels of ADRP,TIP47,JMJD3 and FAS. Except for the same expression level of FAS in different concentration of SAP intervention group,the expression level of FAS increased with the increase of SAP concentration. Conclusion SAP can promote the formation of foam cells and enhance the accumulation of lipid,SAP can enhance the expression of related lipoproteins.
作者
廉会娟
石镜明
崔海燕
李岩松
孙正启
LIAN Huijuan;SHI Jingming;CUI Haiyan;LI Yansong;SUN Zhengqi(Department of Medicine,Tibet Nationalities University,Xianyang,Shaanxi 712082,China)
出处
《中国动脉硬化杂志》
CAS
2018年第9期883-887,共5页
Chinese Journal of Arteriosclerosis
基金
西藏自治区科技厅项目(2016ZR-MQ-04)
西藏民族大学青年学人培育计划项目(16MYQP08)
关键词
淀粉样P物质
泡沫细胞
脂肪分化相关蛋白
serum amyloid P component
foam ceils
adipocyte differentiation related protein
