Cook E B, Stahl J L, Barney N P, Graziano F M
University of Wisconsin-Madison, School of Medicine, Department of Medicine, USA.
Ann Allergy Asthma Immunol. 2001 Nov;87(5):424-9. doi: 10.1016/S1081-1206(10)62926-2.
Olopatadine is a clinically effective dual-action (antihistamine/mast cell stabilizer) ophthalmic antiallergic agent. We have previously demonstrated that olopatadine inhibits tumor necrosis factor alpha (TNF-alpha) release from purified human conjunctival mast cells and that supernates from stimulated mast cells upregulate intercellular adhesion molecule 1 (ICAM-1) expression on epithelial cells via TNF-alpha.
To investigate the effect of olopatadine on the TNF-alpha-mediated mast cell upregulation of ICAM-1 expression on conjunctival epithelial cells.
Human conjunctival mast cells and epithelial cells were purified (>95%) from cadaveric tissue. Conjunctival mast cells were preincubated with three doses (30, 300, or 3,000 microM) of olopatadine or buffer alone for 30 minutes followed by 90-minute challenge with anti-immunoglobulin E (10 microg/mL). The resulting supernates were incubated with conjunctival epithelial cell monolayers for 24 hours along with the following treatments: rTNF-alpha, mast cell supernate + anti-TNF-alpha, recombinant (r)TNF-alpha + anti-TNF-alpha, the three doses of olopatadine, olopatadine supernates, olopatadine supernates + rTNF-alpha. ICAM-1 expression was measured using flow cytometry.
Anti-IgE-stimulated human conjunctival mast cell supernates upregulated human conjunctival epithelial cell ICAM-1 expression to the same extent as rTNF-alpha. ICAM-1 upregulation could be completely blocked with anti-TNF-alpha. Preincubation of conjunctival mast cells with olopatadine significantly blocked the ability of supernates to upregulate ICAM-1 on conjunctival epithelial cells. ICAM-1 expression could be restored by adding rTNF-alpha to the olopatadine-preincubated mast cell supernates.
Olopatadine is able to significantly decrease the anti-immunoglobulin E mast cell supernate-mediated upregulation of ICAM-1 on human conjunctival epithelial cells in vitro. This seems to be mediated through an effect on a TNF-alpha-specific mechanism.
奥洛他定是一种临床有效的双效（抗组胺药/肥大细胞稳定剂）眼科抗过敏药物。我们之前已经证明，奥洛他定可抑制纯化的人结膜肥大细胞释放肿瘤坏死因子α（TNF-α），并且受刺激的肥大细胞的上清液可通过TNF-α上调上皮细胞上细胞间黏附分子1（ICAM-1）的表达。
研究奥洛他定对TNF-α介导的结膜上皮细胞上ICAM-1表达的肥大细胞上调作用的影响。
从尸体组织中纯化人结膜肥大细胞和上皮细胞（>95%）。将结膜肥大细胞分别用三种剂量（30、300或3000 microM）的奥洛他定或单独的缓冲液预孵育30分钟，然后用抗免疫球蛋白E（10 microg/mL）刺激90分钟。将所得上清液与结膜上皮细胞单层一起孵育24小时，并进行以下处理：重组人TNF-α（rTNF-α）、肥大细胞上清液+抗TNF-α、rTNF-α+抗TNF-α、三种剂量的奥洛他定、奥洛他定上清液、奥洛他定上清液+rTNF-α。使用流式细胞术测量ICAM-1的表达。
抗IgE刺激的人结膜肥大细胞上清液上调人结膜上皮细胞ICAM-1表达的程度与rTNF-α相同。抗TNF-α可完全阻断ICAM-1的上调。用奥洛他定预孵育结膜肥大细胞可显著阻断上清液上调结膜上皮细胞上ICAM-1的能力。通过向用奥洛他定预孵育的肥大细胞上清液中添加rTNF-α可恢复ICAM-1的表达。
奥洛他定能够在体外显著降低抗免疫球蛋白E肥大细胞上清液介导的人结膜上皮细胞上ICAM-1的上调。这似乎是通过对TNF-α特异性机制的作用介导的。
