ox-LDL对血小板基质金属蛋白酶诱导剂释放的影响
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全军保健科研基金项目(12BJZ29)


Effect of Oxidized Low Density Lipoprotein on the Release of Extracellular Matrix Metalloproteinase Inducer on Platelets
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    摘要:

    目的 探讨氧化型低密度脂蛋白(ox-LDL)对血小板基质金属蛋白酶诱导剂(EMMPRIN,又名CD147)释放的影响。方法 分别以终浓度25 mg/L、50 mg/L和100 mg/L ox-LDL,对照组(磷酸盐,PBS)和100 mg/L天然LDL体外刺激洗涤血小板,以流式细胞仪检测血小板上CD147、α颗粒膜糖蛋白(CD62P)的表达,酶联免疫吸附法检测血小板上清液中可溶性CD147(sCD147)含量,共聚焦显微镜、透射电子显微镜观察血小板形态改变及颗粒释放。同时,阻断LOX-1受体后,再以流式细胞仪检测ox-LDL(50 mg/L和100 mg/L)刺激的血小板CD147表达。结果 25 mg/L、50 mg/L和100 mg/L ox-LDL处理组血小板CD147相对平均荧光强度(rMFI)(1.01±0.06、1.18±0.07、1.24±0.08)均高于对照组(0.86±0.10, P<0.01)和天然LDL组(0.89±0.11, P<0.01)。LOX-1阻断后,ox-LDL(50 mg/L和100 mg/L)处理组血小板CD147 rMFI(0.96±0.08、1.05±0.07)均下降(P<0.001)。ox-LDL处理后,血小板形态明显改变,并伴随颗粒释放。结论 ox-LDL通过与LOX-1结合诱导了血小板CD147的释放。

    Abstract:

    Aim To investigate the effect of ox-LDL on platelet extracellular matrix metalloproteinase inducer(EMMPRIN,CD147) release. Methods Washed platelets were incubated with ox-LDL (final concentrations of 25 mg/L, 50 mg/L or 100 mg/L ox-LDL, respectively), PBS (as control) or 100 mg/L native LDL in vitro, and the expressions of CD147 and alpha-granule membrane glycoprotein (CD62P) on platelets were detected by flow cytometry. Soluble CD147 from the platelets was assessed by an enzyme-linked immunosorbent assay. Laser scanning microscopy (LSM) and transmission electron microscopy (TEM) were used to visualize the morphological changes and granule release, respectively, from the platelets. In parallel, the expression of CD147 on the platelets pre-incubated with anti-LOX-1 antibody was detected by flow cytometry. Results The CD147 relative mean fluorescence intensity (rMFI) from the groups of 25 mg/L, 50 mg/L or 100 mg/L ox-LDL (1.01±0.06,1.18±0.07, 1.24±0.08, respectively) were higher than that from the control group (0.86±0.10, both P<0.01) or native LDL group (0.89±0.11, both P<0.01). The CD147 rMFI(0.96±0.08,1.05±0.07) from the platelets incubated with ox-LDL (50 mg/L or 100 mg/L) prior to pre-incubation with anti-LOX-1 antibody decreased compared with the 50 mg/L or 100 mg/L ox-LDL-treated platelets, respectively (both P<0.001). After exposure to ox-LDL, morphological changes and granule release in the platelets were visualized by LSM and TEM. Conclusion ox-LDL induces the release of platelet CD147 via binding to LOX-1.

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杨升华,杜大勇,柳 杨,薛 峰,路怀志,张雁斌,王双双,李运田. ox-LDL对血小板基质金属蛋白酶诱导剂释放的影响[J].中国动脉硬化杂志,2014,22(9):907~911.

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  • 收稿日期:2013-10-14
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