PAPP-A通过IGF-1信号途径下调胆固醇流出相关蛋白的表达
发布时间：2018-12-27 16:17
【摘要】：目的：研究显示PAPP-A通过IGF-1激活PI3-K/Akt信号通路发挥促动脉粥样硬化（As）发生发展的作用，此外IGF-1还可通过MAPK信号通路促进As发生发展。本文观察PAPP-A对THP-1巨噬细胞源性泡沫细胞ABCA1、ABCG1和SR-B1表达及其介导胆固醇流出的影响，并探讨其相关机制。 方法：THP-1巨噬细胞源性泡沫细胞经不同浓度PAPP-A（0，10，50，250ng/mL）处理或者用250ng/mL PAPP-A共处理不同时间（0，6，12，24h）;运用液体闪烁计数器检测细胞内胆固醇流出和磷脂流出，高效液相色谱分析细胞内总胆固醇、游离胆固醇和胆固醇酯含量,实时荧光定量PCR和Western blot检测ABCA1、ABCG1、SR-B1和LXRα、IGFBP-5mRNA和蛋白表达情况。以PI3-K抑制剂或MAPK阻断剂或转染IGF-1R siRNA对PAPP-A作用下的细胞共处理24小时，观察对ABCA1、ABCG1、SR-B1及LXRα表达的影响。250ng/mL PAPP-A处理THP-1巨噬细胞源性泡沫细胞24h，ELISA检测培养液中IGF-1的浓度，，实时荧光定量PCR检测细胞内IGF-1mRNA表达情况，Western blot检测PAPP-A对PI3-K和Akt磷酸化水平的影响 结果：PAPP-A抑制ABCA1、ABCG1和SR-B1的表达及其介导的胆固醇流出和磷脂流出；LXRα介导了PAPP-A抑制ABCA1、ABCG1和SR-B1的表达以及细胞内胆固醇流出和磷脂流出；PAPP-A通过IGF-1/PI3K/Akt信号通路抑制LXRα、ABCA1、ABCG1和SR-B1mRNA和蛋白的表达；THP-1巨噬细胞源性泡沫细胞中PAPP-A可激活IGF-1/PI-3K/Akt信号途径。 结论：PAPP-A通过IGF-1/PI3K/Akt信号通路抑制LXRα表达，进而抑制ABCA1、ABCG1和SR-B1表达及细胞内胆固醇流出，从而发挥促As发生发展的作用。
[Abstract]:Aim: to investigate the role of PAPP-A in promoting the development of atherosclerotic (As) through IGF-1 activation of PI3-K/Akt signaling pathway, and IGF-1 can also promote the development of As through MAPK signaling pathway. The effects of PAPP-A on the expression of ABCA1,ABCG1 and SR-B1 in THP-1 macrophage derived foam cells and their effects on cholesterol efflux were investigated. Methods: THP-1 macrophage derived foam cells were treated with different concentrations of PAPP-A (010 ~ 10g / mL) or co-treated with 250ng/mL PAPP-A for 24 h. Cholesterol efflux and phospholipid efflux were detected by liquid scintillation counter, total cholesterol, free cholesterol and cholesterol ester contents were analyzed by HPLC, ABCA1,ABCG1,SR-B1 and LXR 伪 were detected by real-time fluorescence quantitative PCR and Western blot. Expression of IGFBP-5mRNA and protein. The effects of PI3-K inhibitor or MAPK blocker or IGF-1R siRNA transfection on the expression of ABCA1,ABCG1,SR-B1 and LXR 伪 in the cells treated with PAPP-A for 24 hours were observed. 250ng/mL PAPP-A treatment of THP-1 macrophage derived foam cells for 24 h. Detection of IGF-1 concentration in culture medium by ELISA and IGF-1mRNA expression in cells by real-time fluorescence quantitative PCR, Western blot effect of PAPP-A on PI3-K and Akt phosphorylation results: PAPP-A inhibits ABCA1, The expression of ABCG1 and SR-B1 and their mediated cholesterol and phospholipid efflux; LXR 伪 mediated PAPP-A to inhibit the expression of ABCA1,ABCG1, SR-B1, cholesterol and phospholipid, and PAPP-A inhibited the expression of LXR 伪, ABCA1,ABCG1, SR-B1mRNA and protein through IGF-1/PI3K/Akt signaling pathway. PAPP-A can activate IGF-1/PI-3K/Akt signaling pathway in THP-1 macrophage derived foam cells. Conclusion: PAPP-A inhibits the expression of LXR 伪, ABCA1,ABCG1 and SR-B1 and cholesterol efflux through IGF-1/PI3K/Akt signaling pathway, thus promoting the development of As.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363
本文编号：2393305
[Abstract]:Aim: to investigate the role of PAPP-A in promoting the development of atherosclerotic (As) through IGF-1 activation of PI3-K/Akt signaling pathway, and IGF-1 can also promote the development of As through MAPK signaling pathway. The effects of PAPP-A on the expression of ABCA1,ABCG1 and SR-B1 in THP-1 macrophage derived foam cells and their effects on cholesterol efflux were investigated. Methods: THP-1 macrophage derived foam cells were treated with different concentrations of PAPP-A (010 ~ 10g / mL) or co-treated with 250ng/mL PAPP-A for 24 h. Cholesterol efflux and phospholipid efflux were detected by liquid scintillation counter, total cholesterol, free cholesterol and cholesterol ester contents were analyzed by HPLC, ABCA1,ABCG1,SR-B1 and LXR 伪 were detected by real-time fluorescence quantitative PCR and Western blot. Expression of IGFBP-5mRNA and protein. The effects of PI3-K inhibitor or MAPK blocker or IGF-1R siRNA transfection on the expression of ABCA1,ABCG1,SR-B1 and LXR 伪 in the cells treated with PAPP-A for 24 hours were observed. 250ng/mL PAPP-A treatment of THP-1 macrophage derived foam cells for 24 h. Detection of IGF-1 concentration in culture medium by ELISA and IGF-1mRNA expression in cells by real-time fluorescence quantitative PCR, Western blot effect of PAPP-A on PI3-K and Akt phosphorylation results: PAPP-A inhibits ABCA1, The expression of ABCG1 and SR-B1 and their mediated cholesterol and phospholipid efflux; LXR 伪 mediated PAPP-A to inhibit the expression of ABCA1,ABCG1, SR-B1, cholesterol and phospholipid, and PAPP-A inhibited the expression of LXR 伪, ABCA1,ABCG1, SR-B1mRNA and protein through IGF-1/PI3K/Akt signaling pathway. PAPP-A can activate IGF-1/PI-3K/Akt signaling pathway in THP-1 macrophage derived foam cells. Conclusion: PAPP-A inhibits the expression of LXR 伪, ABCA1,ABCG1 and SR-B1 and cholesterol efflux through IGF-1/PI3K/Akt signaling pathway, thus promoting the development of As.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363
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