Massachusetts Eye and Ear Infirmary, Harvard Medical School, Boston, MA.
Department of Biomedical Engineering, Boston University, Boston, MA.
Int Forum Allergy Rhinol. 2015 Jun;5(6):477-80. doi: 10.1002/alr.21454. Epub 2015 Apr 23.
Itraconazole and clarithromycin are clinically effective in the treatment of chronic rhinosinusitis (CRS) through incompletely understood anti-inflammatory properties. P-glycoprotein (P-gp) is overexpressed in CRS and inhibition results in decreased inflammatory cytokine secretion. Both itraconazole and clarithromycin have also been shown to have P-gp inhibitory properties in other tissues, suggesting a novel explanation for their immunomodulatory effects in CRS. The purpose of this study is to therefore confirm whether these drugs are capable of inhibiting P-gp specifically in sinonasal epithelial cells.
This was an institutional review board (IRB)-approved study in which primary sinonasal epithelial cells were cultured in 96-well plates. A Calcein AM assay was used to quantify P-gp inhibition as determined by an increase in intracellular fluorescence. A dose-response curve was generated for itraconazole and clarithromycin (maximal concentration 100 μM) and compared to that of Zosuquidar, a highly specific known P-gp inhibitor. Results were compared using a Student t test with a significance defined as p < 0.05.
Both itraconazole and clarithromycin demonstrated a dose-response curve for P-gp inhibition similar to that of Zosuquidar. The respective maximal inhibitory concentrations of Zosuquidar, itraconazole, and clarithromycin prior to induction of cytotoxicity were 0.31, 3.13, and 1.56 μM, respectively, as demonstrated by a statistically significant increase in total intracellular fluorescence (p < 0.05 in all groups).
Both itraconazole and clarithromycin are capable of inhibiting sinonasal epithelial cell associated P-gp. The anti-inflammatory effects of these agents in CRS may be attributable, in part, to their heretofore unrecognized P-gp modulatory properties.
伊曲康唑和克拉霉素通过不完全了解的抗炎特性在慢性鼻-鼻窦炎(CRS)的治疗中具有临床疗效。多药耐药蛋白(P-gp)在 CRS 中过度表达,抑制 P-gp 可导致炎症细胞因子分泌减少。伊曲康唑和克拉霉素在其他组织中也显示出 P-gp 抑制特性,这表明它们在 CRS 中的免疫调节作用有新的解释。因此,本研究的目的是确认这些药物是否能够特异性抑制鼻黏膜上皮细胞中的 P-gp。
这是一项机构审查委员会(IRB)批准的研究,其中原代鼻黏膜上皮细胞在 96 孔板中培养。使用 Calcein AM 测定法通过增加细胞内荧光来定量 P-gp 抑制。生成伊曲康唑和克拉霉素(最大浓度 100 μM)的剂量反应曲线,并与高度特异性已知 P-gp 抑制剂唑吡坦进行比较。使用学生 t 检验比较结果,以 p < 0.05 为显著性定义。
伊曲康唑和克拉霉素均显示出与唑吡坦相似的 P-gp 抑制剂量反应曲线。在诱导细胞毒性之前,唑吡坦、伊曲康唑和克拉霉素的最大抑制浓度分别为 0.31、3.13 和 1.56 μM,这导致总细胞内荧光显著增加(所有组均 p < 0.05)。
伊曲康唑和克拉霉素均能抑制鼻黏膜上皮细胞相关的 P-gp。这些药物在 CRS 中的抗炎作用可能部分归因于它们以前未被认识到的 P-gp 调节特性。