Reduced beta2-glycoprotein I protects macrophages from ox-LDL-induced foam cell formation and cell apoptosis.
Reduced beta2-glycoprotein I protects macrophages from ox-LDL-induced foam cell formation and cell apoptosis.
复制标题
还原的 β2-糖蛋白 I 可保护巨噬细胞免受 ox-LDL 诱导的泡沫细胞形成和细胞凋亡的影响。
DOI：
10.1186/1476-511x-12-174
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发表时间：
2013-11-16
影响因子：
4.5
通讯作者：
Yu P
中科院分区：
文献类型：
--
作者：
Wang WL;Meng ZX;Zhou SJ;Li CJ;Chen R;Lv L;Ma ZJ;Yu DM;Yu P
还原型β 2-糖蛋白I（β 2-GPI）是β 2-GPI的一种含巯基的游离形式，在保护内皮细胞免受氧化应激诱导的细胞死亡方面显示出强大的作用。本研究旨在研究β 2-GPI或还原型β 2-GPI对ox-LDL诱导的泡沫细胞形成和细胞凋亡的影响，并确定可能的机制。以RAW264.7巨噬细胞系为实验材料。油红O染色法定性检测胆固醇蓄积，胆固醇测定法定量检测胆固醇蓄积。流式细胞术检测细胞凋亡。采用实时定量PCR方法检测与胆固醇转运相关的主要蛋白CD 36、SRB 1、ABCA 1和ABCG 1的mRNA表达。Western blot检测caspase-9、caspase-3、p38 MAPK/p-p38 MAPK、JNK/p-JNK等凋亡相关蛋白的表达。β 2-GPI或降低β 2-GPI可降低ox-LDL诱导的胆固醇蓄积（96.45 ± 8.51 μg/mg蛋白vs 114.35 ± 10.38 μg/mg蛋白，p < 0.05; 74.44 ± 5.27 μg/mg蛋白vs114.35 ± 10.38 μg/mg蛋白，P < 0.01）和细胞凋亡（30.00 ± 5.10%vs.38.70 ± 7.76%，p < 0.05; 20.66 ± 2.50%vs.38.70 ± 7.76%，p < 0.01），且β 2-GPI与降低的β 2-GPI之间存在显著差异（p < 0.05）。降低的β 2-GPI降低了ox-LDL诱导的CD 36 mRNA和ABCA 1 mRNA的表达（p < 0.05），以及CD 36、裂解的caspase-9、裂解的caspase-3、p-p38 MAPK和p-JNK蛋白的表达（p < 0.05或p < 0.01）。β 2-GPI对ABCA 1 mRNA和p-p38 MAPK蛋白表达无明显影响。β 2-GPI和还原型β 2-GPI均能抑制ox-LDL诱导的泡沫细胞形成和细胞凋亡，还原型β 2-GPI的抑制作用更强。这两种糖蛋白通过下调CD 36以及蛋白质表达来减少巨噬细胞的脂质摄入。降低的β 2-GPI通过减少ox-LDL诱导的p38 MAPK和JNK磷酸化以及切割的caspase-3和caspase-9的量来抑制细胞凋亡。β 2-GPI不抑制ox-LDL诱导的p38 MAPK磷酸化。
Reduced beta2-glycoprotein I (beta2-GPI) is a free thiol-containing form of beta2-GPI that displays a powerful effect in protecting endothelial cells from oxidative stress-induced cell death. The present study aims to investigate the effect of beta2-GPI or reduced beta2-GPI on ox-LDL-induced foam cell formation and on cell apoptosis and to determine the possible mechanisms. The RAW264.7 macrophage cell line was selected as the experimental material. Oil red O staining and cholesterol measurement were used to detect cholesterol accumulation qualitatively and quantitatively, respectively. Flow cytometry was used to detect cell apoptosis. Real-time quantitative PCR was used to detect the mRNA expression of the main proteins that are associated with the transport of cholesterol, such as CD36, SRB1, ABCA1 and ABCG1. Western blot analysis was used to detect the protein expression of certain apoptosis-related proteins, such as caspase-9, caspase-3, p38 MAPK/p-p38 MAPK and JNK/p-JNK. Beta2-GPI or reduced beta2-GPI decreased ox-LDL-induced cholesterol accumulation (96.45 ± 8.51 μg/mg protein vs. 114.35 ± 10.38 μg/mg protein, p < 0.05;74.44 ± 5.27 μg/mg protein vs. 114.35 ± 10.38 μg/mg protein, p < 0.01) and cell apoptosis (30.00 ± 5.10% vs. 38.70 ± 7.76%, p < 0.05; 20.66 ± 2.50% vs. 38.70 ± 7.76%, p < 0.01), and there are significant differences between beta2-GPI and reduced beta2-GPI (p < 0.05). Reduced beta2-GPI decreased the ox-LDL-induced expression of CD36 mRNA and ABCA1 mRNA (p < 0.05), as well as CD36, cleaved caspase-9, cleaved caspase-3, p-p38 MAPK and p-JNK proteins (p < 0.05 or p < 0.01). Beta2-GPI did not significantly decrease the expression of ABCA1 mRNA and the p-p38 MAPK protein. Both beta2-GPI and reduced beta2-GPI inhibit ox-LDL-induced foam cell formation and cell apoptosis, and the latter exhibits a stronger inhibition effect. Both of these glycoproteins reduce the lipid intake of macrophages by downregulating CD36 as well as protein expression. Reduced beta2-GPI inhibits cell apoptosis by reducing the ox-LDL-induced phosphorylation of p38 MAPK and JNK, and the amount of cleaved caspase-3 and caspase-9. Beta2-GPI does not inhibit the ox-LDL-induced phosphorylation of p38 MAPK.