德尔塔阿片受体可减弱 TNF-α 诱导的人视盘神经节星形胶质细胞 MMP-2 的分泌。
Delta-opioid receptors attenuate TNF-α-induced MMP-2 secretion from human ONH astrocytes.
机构信息
Hewitt Laboratory of the Ola B. Williams Glaucoma Center, Department of Ophthalmology, Storm Eye Institute, Medical University of South Carolina, Charleston, South Carolina.
出版信息
Invest Ophthalmol Vis Sci. 2013 Oct 9;54(10):6605-11. doi: 10.1167/iovs.13-12196.
PURPOSE
We examined the signaling mechanisms involved in δ-opioid-receptor agonist, SNC-121-mediated attenuation of TNF-α-induced matrix metalloproteinase-2 (MMP-2) secretion from human optic nerve head (ONH) astrocytes.
METHODS
Human ONH astrocytes were treated with SNC-121 (1 μmol/L) for 15 minutes followed by TNF-α (25 ng/mL) treatment for 6 or 24 hours. Cells were pretreated with inhibitors of p38 mitogen-activated protein (MAP) kinase (SB-203580) or NF-κB (Helenalin) prior to TNF-α treatment. Changes in phosphorylation and expression of p38 MAP kinase, IκBα, NF-κB, and MMP-2 were measured by Western blotting. Translocation of NF-κB was determined by immunocytochemistry.
RESULTS
TNF-α treatment increased MMP-2 secretion from ONH astrocytes to 236% ± 17% and 142% ± 8% at 6 and 24 hours, respectively; while SNC-121 treatment reduced MMP-2 secretion to 149% ± 11% and 108% ± 7% at 6 and 24 hours, respectively. The SNC-121-mediated inhibitory response was blocked by the δ-opioid-receptor antagonist naltrindole. TNF-α treatment resulted in a sustained phosphorylation of p38 MAP kinase up to 24 hours (226% ± 15% over control levels), which was reduced to 150% ± 20% by SNC-121 treatment. TNF-α treatment increased the expression of NF-κB to 179% ± 21% and 139% ± 6% at 6 and 24 hours, respectively, which was significantly blocked by SNC-121 treatment. Furthermore, TNF-α-induced MMP-2 secretion was blocked by 100% and 78% in the presence of SB-203580 and Helenalin, respectively.
CONCLUSIONS
Evidence is provided that SNC-121 attenuated TNF-α-induced MMP-2 secretion from ONH astrocytes. Data also supported the idea that p38 MAP kinase and NF-κB played central roles in TNF-α-induced MMP-2 secretion, and both were negatively regulated by SNC-121.
目的
我们研究了 δ 阿片受体激动剂 SNC-121 抑制 TNF-α 诱导的人视神经头（ONH）星形胶质细胞基质金属蛋白酶-2（MMP-2）分泌所涉及的信号转导机制。
方法
用 SNC-121（1 μmol/L）处理人 ONH 星形胶质细胞 15 分钟，然后用 TNF-α（25ng/mL）处理 6 或 24 小时。用 p38 丝裂原活化蛋白（MAP）激酶（SB-203580）或 NF-κB（Helenalin）抑制剂预处理细胞，然后用 TNF-α 处理。通过 Western 印迹法测定 p38 MAP 激酶、IκBα、NF-κB 和 MMP-2 的磷酸化和表达变化。通过免疫细胞化学测定 NF-κB 的易位。
结果
TNF-α 处理使 ONH 星形胶质细胞的 MMP-2 分泌分别增加至 236%±17%和 142%±8%，分别在 6 和 24 小时；而 SNC-121 处理使 MMP-2 分泌分别减少至 149%±11%和 108%±7%，分别在 6 和 24 小时。δ 阿片受体拮抗剂纳曲吲哚阻断 SNC-121 介导的抑制反应。TNF-α 处理导致 p38 MAP 激酶的磷酸化持续至 24 小时（与对照水平相比增加 226%±15%），而 SNC-121 处理则减少至 150%±20%。TNF-α 处理使 NF-κB 的表达分别增加至 179%±21%和 139%±6%，分别在 6 和 24 小时，而 SNC-121 处理则显著阻断 NF-κB 的表达。此外，在存在 SB-203580 和 Helenalin 的情况下，TNF-α 诱导的 MMP-2 分泌分别被阻断 100%和 78%。
结论
证据表明，SNC-121 减轻了 TNF-α 诱导的 ONH 星形胶质细胞 MMP-2 分泌。数据还支持 p38 MAP 激酶和 NF-κB 在 TNF-α 诱导的 MMP-2 分泌中起核心作用的观点，并且两者均受 SNC-121 的负调控。
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