Ghosh Abhijit, Pechota L V T Angela, Upchurch Gilbert R, Eliason Jonathan L
Section of Vascular Surgery, Department of Surgery, Jobst Vascular Research Laboratories, University of Michigan Medical School, Ann Arbor, MI, USA.
Division of Vascular and Endovascular Surgery, University of Virginia, PO Box 800679, Charlottesville, VA, USA.
Mol Cell Biochem. 2015 Dec;410(1-2):75-84. doi: 10.1007/s11010-015-2539-3. Epub 2015 Aug 29.
We hypothesized that matrix metalloproteinase secretion in response to cigarette smoke is modulated by cross-talk between resident cells within the aorta, namely, aortic smooth muscles, endothelial cells, and infiltrating macrophages, and this may be crucial for in vivo formation/progression of abdominal aortic aneurysm (AAA). Cigarette smoke extract (CSE) was applied to rat aortic smooth muscle (RASMC), endothelial (RAEC) or RAW cells, and conditioned media (CSE-CM) collected. Fresh cells were treated with CSE-CM for 24 h and then maintained in serum-free medium (SFM) for 72 h to analyze MMP2 and MMP9 in media by zymography and the ratio (pS/pJ) of phospho-Stat3 (pStat3) and phospho-Jak2 (pJak2) inside the cells by Western blot. We observed that CSE-CM from RAW and RAEC increased MMP9 by 200 and 17 %, respectively, in RASMC and also increased pS/pJ ratio (305 and 228 %, respectively) in RASMC. RAW cell-derived CSE-CM induced RAEC to produce moderate amounts of MMP2 (17 %), MMP9 (30 %), and a 137 % increase in pS/pJ. RAW cells receiving unstimulated CM from RASMC and RAEC produced significant amounts of MMP9 (128 and 155 %, respectively) and increased pS/pJ (45 and 1283 %, respectively). CSE-CM from RASMC and RAEC induced significant production of MMP9 from RAW cells (237 and 162 %, respectively) and increase in pS/pJ ratios (1348 and 1494 %, respectively). This is the first in vitro study demonstrating cigarette smoke extract-mediated differential interactions between resident cells in the aorta leads to altered modulation of signaling molecules that may be vital for AAA formation under in vivo conditions.
我们推测，主动脉内的驻留细胞（即主动脉平滑肌细胞、内皮细胞和浸润的巨噬细胞）之间的相互作用调节了对香烟烟雾产生反应时基质金属蛋白酶的分泌，这可能对腹主动脉瘤（AAA）的体内形成/进展至关重要。将香烟烟雾提取物（CSE）应用于大鼠主动脉平滑肌（RASMC）、内皮（RAEC）或RAW细胞，并收集条件培养基（CSE-CM）。将新鲜细胞用CSE-CM处理24小时，然后在无血清培养基（SFM）中维持72小时，通过酶谱分析培养基中的MMP2和MMP9，并通过蛋白质印迹法分析细胞内磷酸化Stat3（pStat3）和磷酸化Jak2（pJak2）的比率（pS/pJ）。我们观察到，RAW和RAEC的CSE-CM分别使RASMC中的MMP9增加了200%和17%，并且还使RASMC中的pS/pJ比率分别增加了305%和228%。RAW细胞来源的CSE-CM诱导RAEC产生适量的MMP2（17%）、MMP9（30%），并且pS/pJ增加了137%。接受来自RASMC和RAEC的未刺激CM的RAW细胞产生了大量的MMP9（分别为128%和155%），并且pS/pJ增加（分别为45%和1283%）。RASMC和RAEC的CSE-CM诱导RAW细胞产生大量的MMP9（分别为237%和162%），并且pS/pJ比率增加（分别为1348%和1494%）。这是第一项体外研究，表明香烟烟雾提取物介导的主动脉驻留细胞之间的差异相互作用导致信号分子的调节改变，这可能对体内条件下AAA的形成至关重要。
