Department of Biochemistry and Molecular Biology, University of Bari, Bari, Italy.
PLoS One. 2011;6(12):e28499. doi: 10.1371/journal.pone.0028499. Epub 2011 Dec 8.
Matrix metalloproteinases (MMPs) released by glial cells are important mediators of neuroinflammation and neurologic damage in HIV infection. The use of antiretroviral drugs able to combat the detrimental effect of chronic inflammation and target the exaggerated MMP activity might represent an attractive therapeutic challenge. Recent studies suggest that CCR5 antagonist maraviroc (MVC) exerts immunomodulant and anti-inflammatory activity beyond its anti-HIV properties. We investigated the in vitro effect of MVC on the activity of MMPs in astrocyte and microglia cultures.
METHODOLOGY/PRINCIPAL FINDINGS: Primary cultures of rat astrocytes and microglia were activated by exposure to phorbol myristate acetate (PMA) or lypopolysaccharide (LPS) and treated in vitro with MVC. Culture supernatants were subjected to gelatin zymography and quantitative determination of MMP-9 and MMP-2 was done by computerized scanning densitometry. MMP-9 levels were significantly elevated in culture supernatants from both LPS- and PMA-activated astrocytes and microglia in comparison to controls. The treatment with MVC significantly inhibited in a dose-dependent manner the levels and expression of MMP-9 in PMA-activated astrocytes (p<0,05) and, to a lesser extent, in PMA-activated microglia. By contrast, levels of MMP-2 did not significantly change, although a tendency to decrease was seen in PMA-activated astrocytes after treatment with MVC. The inhibition of levels and expression of MMP-9 in PMA-activated glial cells did not depend on cytotoxic effects of MVC. No inhibition of MMP-9 and MMP-2 were found in both LPS-activated astrocytes and microglia.
The present in vitro study suggests that CCR5 antagonist compounds, through their ability to inhibit MMP-9 expression and levels, might have a great potential for the treatment of HIV-associated neurologic damage.
胶质细胞释放的基质金属蛋白酶（MMPs）是 HIV 感染中神经炎症和神经损伤的重要介质。使用能够对抗慢性炎症的有害影响并针对 MMP 活性过度的抗逆转录病毒药物可能代表一种有吸引力的治疗挑战。最近的研究表明，CCR5 拮抗剂马拉维若（MVC）除了具有抗 HIV 特性外，还具有免疫调节和抗炎作用。我们研究了 MVC 对星形胶质细胞和小胶质细胞培养物中 MMP 活性的体外影响。
方法/主要发现：用佛波醇肉豆蔻酸酯（PMA）或脂多糖（LPS）激活大鼠星形胶质细胞和小胶质细胞的原代培养物，并在体外用 MVC 处理。培养上清液进行明胶酶谱分析，并通过计算机扫描密度法定量测定 MMP-9 和 MMP-2。与对照组相比，LPS 和 PMA 激活的星形胶质细胞和小胶质细胞培养上清液中 MMP-9 水平明显升高。MVC 以剂量依赖性方式显著抑制 PMA 激活的星形胶质细胞（p<0.05）和 PMA 激活的小胶质细胞中 MMP-9 的水平和表达，抑制作用较弱。相比之下，MMP-2 的水平没有明显变化，尽管在用 MVC 处理后 PMA 激活的星形胶质细胞中观察到下降的趋势。PMA 激活的神经胶质细胞中 MMP-9 水平和表达的抑制与 MVC 的细胞毒性作用无关。在 LPS 激活的星形胶质细胞和小胶质细胞中均未发现 MMP-9 和 MMP-2 的抑制。
本体外研究表明，CCR5 拮抗剂化合物通过抑制 MMP-9 的表达和水平，可能具有治疗 HIV 相关神经损伤的巨大潜力。
