内毒素对THP-1源性巨噬细胞PCSK9和LDLR表达的影响

发布时间：2018-03-30 09:35

本文选题：内毒素　切入点：THP-1源性巨噬细胞　出处：《南华大学》2012年硕士论文

【摘要】：目的以THP-1源性巨噬细胞为研究对象，经内毒素处理后，观察细胞水平PCSK9、LDLR的变化及其规律，，以便了解PCSK9是否介导内毒素在动脉粥样硬化发展中起作用。方法用不同浓度的内毒素(0,0.1,1,10,100μg/ml)处理经160nmol/L佛波酯诱导贴壁的THP-1源性巨噬细胞不同时间(0,3,6,12,24h)，RT-PCR、Western blot分别检测PCSK9、LDLR mRNA和蛋白质的表达。应用Lipofectamine2000转染80nmol/L PCSK9siRNA进入THP-1源性巨噬细胞后，Western blot检测LDLR蛋白质的表达以及油红O染色观察细胞内脂质含量。结果随着内毒素浓度的增加，THP-1源性巨噬细胞PCSK9mRNA、蛋白质的表达逐渐增多。但是随着内毒素浓度的增加，THP-1源性巨噬细胞LDLR蛋白质表达逐渐减少；RT-PCR检测发现，LDLR mRNA表达在早期时间(3h和6h)减少，但随后(12h和24h) LDLR mRNA表达逐渐增多，到24h表达量略多于对照组。PCSK9siRNA转染THP-1源性巨噬细胞后，与LPS单独处理组相比，PCSK9siRNA组、PCSK9siRNA+LPS组LDLR蛋白质表达都增多，PCSK9siRNA组表达最为明显，其细胞内脂质的含量最多。结论 1.内毒素上调THP-1源性巨噬细胞PCSK9mRNA、蛋白质的表达；下调THP-1源性巨噬细胞LDLR蛋白质的表达，而对于LDLR mRNA的表达是可变化的； 2.内毒素通过PCSK9下调THP-1源性巨噬细胞LDLR的表达，影响脂质的蓄积。
[Abstract]:Purpose. The changes and regularity of THP-1 derived macrophages were observed after endotoxin treatment in order to understand whether endotoxin may play a role in the development of atherosclerosis. Method. THP-1 derived macrophages were treated with different concentrations of endotoxin 0. 1 ~ (1) ~ 1 ~ (10) 渭 g 路ml ~ (-1)) to observe the expression of mRNA and protein in THP-1 derived macrophages induced by 160nmol/L phorbol ester at different times. The expression of mRNA and protein in THP-1 derived macrophages was detected by Lipofectamine2000 transfection of 80nmol/L PCSK9siRNA into THP-1 derived macrophages. The expression of LDLR protein and the lipid content in the cells were observed by oil red O staining. Results. With the increase of endotoxin concentration, the protein expression of THP-1 derived macrophage PCSK9mRNAs gradually increased, but with the increase of endotoxin concentration, the LDLR protein expression of THP-1 derived macrophages decreased gradually. The period time of 3 h and 6 h) decreased, However, the expression of LDLR mRNA increased gradually at 12h and 24h. After transfection of PCSK9siRNA into THP-1 derived macrophages at 24 h, the expression of LDLR protein in PCSK9siRNA group was higher than that in PCSK9siRNA LPS group, and the expression of LDLR protein in PCSK9siRNA group was higher than that in PCSK9siRNA group. The content of lipid in the cells was the highest. Conclusion. 1. Endotoxin up-regulated the expression of PCSK9mRNAs in THP-1 derived macrophages, down-regulated the expression of LDLR protein in THP-1 derived macrophages, and changed the expression of LDLR mRNA. 2. Endotoxin down-regulated the expression of LDLR in THP-1 derived macrophages by PCSK9, and affected the accumulation of lipids.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363

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