섬유주세포에서 항혈관내피성장인자가 기질금속단백분해효소의 활성에 미치는 영향

Effect of Anti-vascular Endothelial Growth Factor on the Matrix Metalloproteinase Activity in Trabecular Meshwork Cells

목적: 섬유주세포에서 항혈관내피성장인자(anti-vascular endothelial growth factor, anti-VEGF)가 기질금속단백분해효소(matrix metalloproteinase, MMP)의 활성에 미치는 영향을 알아보고자 하였다. 대상과 방법: 인체의 섬유주세포를 배양한 다음 0.25, 0.50 mg/mL anti-VEGF bevacizumab (BV)을 24시간 노출시킨 다음, carboxyfluorescein을 이용하여 섬유주단층세포층의 투과도를 측정하였고 trans-epithelial endothelial electrical resistance (TEER)를 측정하였다. Western blot으로 MMP-1/-2 단백의 활성을 측정하였으며 Griess assay를 이용하여 일산화질소(nitric oxide, NO)의 생성 정도를 측정하였다. 결과: 0.50 mg/mL BV은 carboxyfluorescein의 섬유주단층세포층의 투과도를 유의하게 감소시켰으며(p=0.017) TEER에는 유의한 영향을 미치지 않았다(p=0.308). 0.50 mg/mL BV은 MMP-1/-2 단백의 활성을 각각 유의하게 감소시켰으며(p=0.014, p=0.016), 0.50 mg/mL BV은 NO의 생성을 유의하게 감소시켰다(p=0.023). 결론: Anti-VEGF는 섬유주단층세포층의 투과도를 감소시키며, 이와 동반하여 MMP-1/-2 단백의 활성과 NO의 생성을 감소시키는 작용을 나타낸다.

Purpose: To investigate the effects of anti-vascular endothelial growth factor (anti-VEGF) on the activity of matrix metalloproteinase (MMP) in human trabecular meshwork cells (HTMC). Methods: Primary HTMC cultures were exposed to 0, 0.25, and 0.50 mg/mL anti-VEGF bevacizumab (BV) for 24 hours. The permeability through the trabecular meshwork cell monolayer was assessed using carboxyfluorescein and trans-epithelial endothelial electrical resistance (TEER). The levels of MMP-1/-2 were measured by Western blotting and the production of nitric oxide (NO) was assessed with the Griess assay. Results: Bevacizumab at 0.50 mg/mL decreased the permeability of carboxyfluorescein significantly (p = 0.017) and did not affect TEER (p = 0.308). Administration of 0.50 mg/mL BV decreased MMP-1 and MMP-2 activities (p = 0.014, p = 0.016, respectively) and inhibited NO production significantly (p = 0.023). Conclusions: Anti-VEGF decreased the permeability through the HTMC monolayer, which was accompanied by decreased MMP-1/-2 activity and limited NO production.