Salmeterol Xinafoate (GR 33343X xinafoate) is a long-acting β2-adrenergic receptor agonist with anti-inflammatory effects, used in the treatment of asthma symptoms and chronic obstructive pulmonary disease (COPD) symptoms.

β2-adrenoceptor (WT):Ki:1.5 nM

Salmeterol 显著抑制RAW264.7和THP-1细胞产生的促炎介质。Salmeterol 降低PgLPS介导的ERK1/2和JNK的磷酸化，但不影响p38 MAP激酶（MAP-K）。Salmeterol 通过抑制p65-NFκB的核内转移、NF-κB的转录活性及IκBα的磷酸化，减弱NF-κB的激活。Salmeterol 对WT β2AR（β1 Ki /β2 Ki比率约为1500）具有很高的选择性，Ki值为1.5±0.4 nM。Salmeterol 阻止由肿瘤坏死因子-α引起的IRS-1Ser307的磷酸化水平。Salmeterol 单独使用可以防止视网膜Müller细胞死亡（与25 mM葡萄糖相比，p<0.05）。与IRS-1 shRNA联合使用时，相比于仅使用Salmeterol ，细胞死亡率显著增加。此外，仅Salmeterol 处理显著减少细胞色素C水平，当与IRS-1 shRNA联合使用时，此效果减弱。Salmeterol （100 μM）导致树突细胞（DCs）凋亡，但在10 μM浓度下不影响DCs的分化和成熟。Salmeterol （10 μM）降低LPS激活的DCs中促炎细胞因子的mRNA和蛋白水平，抑制MAPK和NF-κB激活。

OVA/LPS组合与salmeterol联用可显著且剂量依赖性地减少过敏小鼠增强的AHR。通过调节小鼠气道炎症，salmeterol有效对抗哮喘。

The cells are rinsed twice with ice-cold phosphate-buffered saline and mechanically detached in ice-cold buffer containing 10 mM Tris·HCl, pH 7.4, 5 mM EDTA, 10 μg/mL benzamidine, 10 μg/mL soybean trypsin inhibitor (type II-S), and 5 μg/mL leupeptin (lysis buffer). The lysate is centrifuged at 45,000 ×g for 10 min at 4°C. The pellet is rehomogenized in lysis buffer, with a Potter-type homogenizer, and stored at ?80°C until use. The competition binding assays are performed in buffer containing 75 mM Tris·HCl, pH 7.4, 12.5 mM MgCl2, and 2 mM EDTA, using 1-5 μg of membrane protein, 50 pM 125I-CYP, and 0-100 μM unlabeled ligand in the presence of 100 μM GTP, for 60 min at 37°C. The binding reaction is terminated by dilution and rapid filtration through Whatman GF/C filters; the filters are washed three times with solution containing 25 mM Tris·HCl, pH 7.4, and 1 mM MgCl2. Nonspecific binding is determined in the presence of 5 μM (±)-propranolol. The radioactivity on the filters is counted with a γ-counte

Salmeterol is formulated in PBS.All mice are sensitized on days 0 and 14 by intraperitoneal injection of either PBS or 0.08 mg OVA and 0.1 mL aluminum hydroxide in 0.1 mL of PBS (pH 7.4). After sensitization, animals are exposed to aerosolized PBS-only (negative control), 1% OVS/PBS (acute exposure), 1% OVA/0.01% LPS/PBS (extra LPS exposure) or 1% OVA/0.01% LPS/salmeterol/PBS (sal treatment) for 40 min, once per day for 3 consecutive days (days 24-26). On day 27, the mice are killed and lungs are divided into two groups for analysis: the left lung lobes are lavaged three times with 1 mL of PBS with 1% fetal calf serum and 5 U/mL heparin, and the right halves are fixed by 4% paraformaldehyde for histological analysis.