Yu Chen, Lixia Yang, Ruiwei Guo, Yankun Shi, Jinshan Ye
Department of Postgraduate, Third Military Medical University (Army Medical University).
Department of Cardiology, Kunming General Hospital of Chengdu Military Command.
Int Heart J. 2018 Mar 30;59(2):394-398. doi: 10.1536/ihj.17-221. Epub 2018 Mar 20.
To investigate whether focal adhesion kinase (FAK) can participate in the secretion of matrix metalloproteinase 9 (MMP9) after CD147 stimulation in THP-1 induced macrophages; thus, to explore the potential treatment perspectives for acute coronary syndrome (ACS).Phorbol-12-myristate-13-acetate (PMA) was used to induce THP-1 cells to differentiate into macrophages. To confirm the peak mRNA and protein expression of FAK and MMP9 after the stimulation of CD147, the macrophages were divided into 5 groups (0, 3, 6, 9, and 12 hours), with 0 hours group as control group. To investigate the role of FAK in the secretion of MMP9, with stimulation of CD147 for 9 hours, FAK inhibitor 14 was used to inhibit FAK Y397 phosphorylation. The mRNA and protein expressions were quantified by qRT-PCR and western blotting, respectively. (1) Relative mRNA expression of FAK and MMP9 were both significantly up-regulated (all P < 0.05) after stimulation of CD147, FAK peaked at 9 hours (3.908 ± 0.106 versus 1, P < 0.05), whereas MMP9 peaked at 6 hours (2.522 ± 0.062 versus 1, P < 0.05). (2) Relative protein expression of FAK, pFAK, and MMP9 were all significantly increased after CD147 stimulation (all P < 0.05), FAK (1.930 ± 0.024 versus 1, P < 0.05) and pFAK (1.737 ± 0.021 versus 1, P < 0.05) peaked at 9 hours, whereas MMP9 peaked at 6 hours (1.527 ± 0.033 versus 1, P < 0.05). (3) CD147 up-regulates FAK, pFAK, and MMP9 mRNA and protein expressions in a dose-dependent manner. (4) FAK inhibitor 14 significantly reduced the relative protein expression level of pFAK (0.077 ± 0.012 versus 1, P < 0.05) and MMP9 (0.133 ± 0.012) at 9 hours after CD147 stimulation.The results demonstrated that FAK Y397 phosphorylation was involved in the secretion of MMP9 after CD147 stimulation in macrophages and may play a role in the regulation of ACS.
探讨在CD147刺激THP-1诱导的巨噬细胞后，粘着斑激酶（FAK）是否能参与基质金属蛋白酶9（MMP9）的分泌；从而探索急性冠状动脉综合征（ACS）的潜在治疗前景。用佛波酯（PMA）诱导THP-1细胞分化为巨噬细胞。为确定CD147刺激后FAK和MMP9的mRNA和蛋白表达峰值，将巨噬细胞分为5组（0、3、6、9和12小时），以0小时组作为对照组。为研究FAK在MMP9分泌中的作用，在CD147刺激9小时后，用FAK抑制剂14抑制FAK Y397磷酸化。分别通过qRT-PCR和蛋白质免疫印迹法对mRNA和蛋白表达进行定量分析。（1）CD147刺激后FAK和MMP9的相对mRNA表达均显著上调（均P<0.05），FAK在9小时达到峰值（3.908±0.106比1，P<0.05），而MMP9在6小时达到峰值（2.522±0.062比1，P<0.05）。（2）CD147刺激后FAK、pFAK和MMP9的相对蛋白表达均显著增加（均P<0.05），FAK（1.930±0.024比1，P<0.05）和pFAK（1.737±0.021比1，P<0.05）在9小时达到峰值，而MMP9在6小时达到峰值（1.527±0.033比1，P<0.05）。（3）CD147以剂量依赖性方式上调FAK、pFAK和MMP9的mRNA和蛋白表达。（4）在CD147刺激9小时后，FAK抑制剂14显著降低了pFAK（0.077±0.012比1，P<0.05）和MMP9（0.133±0.012）的相对蛋白表达水平。结果表明，FAK Y397磷酸化参与了巨噬细胞在CD147刺激后MMP9的分泌，可能在ACS的调控中发挥作用。
