Trybus E, Krol G, Obarzanowski T, Trybus W, Kopacz-Bednarska A, Obarzanowski M, Krol T
Department of Cell Biology and Electron Microscopy, Jan Kochanowski University, Kielce, Poland.
Faculty of Management, University of Warsaw, Warsaw, Poland.
J Physiol Pharmacol. 2017 Dec;68(6):907-919.
Most studies on the effects of glucocorticosteroid therapy in rhinitis relate to their inhibitory effect on activation and the number of inflowing cells that are involved in the development and maintenance of inflammation. It is also very important to determine the range of effect of budesonide on residing cells (epithelial cells). The purpose of this study was to evaluate the effect of local budesonide therapy on the cytological image of the nasal mucosa, with attention paid to columnar cells in patients with rhinitis. The in vivo results obtained were analyzed in correlation with changes in normal CHO-K1 cells exposed to budesonide at concentrations falling within the pharmacological dose range. Fifty patients diagnosed with rhinitis with suspected allergic background without nasal polyps were included in clinical trials. The control group were 10 healthy people without clinical signs of rhinitis. Only in patients with homogeneous cytological picture, exfoliative cytology was performed before treatment and after 4 weeks of therapy with budesonide used in aerosol form. Papanicolaou and Pappenheim - stained smears were evaluated qualitatively and quantitatively for changes in nasal mucosal cells. The nasal mucosal image of the patients before treatment clearly indicated the pathological state confirmed by the presence of numerous neutrophils, eosinophils, abundant bacterial flora and goblet or epithelial cells prevalence. In contrast, in smears of patients post-treatment budesonide observed a clear improvement in their nasal mucosa by reducing inflammation. There was a significant increase in the number of columnar cells and the appearance of very numerous epithelial cells with increased cytoplasmic vacuolization and visible leucophagocytosis. In vitro studies were performed on normal CHO-K1 cells that were treated with budesonide at concentrations of 0.5 μM - 45 μM. After 48 hours of incubation with the test agent, the samples were prepared for optical microscopy using the H&E method and transmission electron microscopy. Comparison of cells exposed to budesonide with control cells (without addition of test agent) revealed vacuolization changes with autophagy. Apoptotic changes have also been demonstrated, which occured to a lesser extent than vacuolization. The changes observed after budesonide treatment in the cytological picture of patients with allergic rhinitis indicate the therapeutic effect of this drug. On the other hand, the changes observed in the cytoplasm of epithelial cells, such as autophagy (clearly promoted in CHO-K1 cells) and leucophagocytosis, may indicate an additional mechanism of action for budesonide.
大多数关于糖皮质激素疗法对鼻炎影响的研究都涉及其对参与炎症发生和维持的细胞活化及流入数量的抑制作用。确定布地奈德对驻留细胞（上皮细胞）的作用范围也非常重要。本研究的目的是评估局部布地奈德疗法对鼻黏膜细胞学图像的影响，重点关注鼻炎患者的柱状细胞。将获得的体内结果与在药理剂量范围内暴露于布地奈德的正常CHO-K1细胞的变化进行相关性分析。五十名被诊断为疑似过敏性背景且无鼻息肉的鼻炎患者被纳入临床试验。对照组为10名无鼻炎临床症状的健康人。仅对细胞学图像均匀的患者，在治疗前以及使用气雾剂形式的布地奈德治疗4周后进行脱落细胞学检查。对巴氏染色和帕彭海姆染色的涂片进行定性和定量评估，以观察鼻黏膜细胞的变化。治疗前患者的鼻黏膜图像清楚地显示出病理状态，表现为存在大量中性粒细胞、嗜酸性粒细胞、丰富的细菌菌群以及杯状或上皮细胞占优势。相比之下，在接受布地奈德治疗后的患者涂片中，观察到鼻黏膜炎症减轻有明显改善。柱状细胞数量显著增加，出现大量上皮细胞，细胞质空泡化增加且可见白细胞吞噬现象。对正常CHO-K1细胞进行体外研究，用浓度为0.5μM - 45μM的布地奈德处理。与测试剂孵育48小时后，使用苏木精和伊红（H&E）方法以及透射电子显微镜对样品进行光学显微镜检查准备。将暴露于布地奈德的细胞与对照细胞（未添加测试剂）进行比较，发现有自噬引起的空泡化变化。还证实了凋亡变化，但其发生程度小于空泡化。布地奈德治疗后在过敏性鼻炎患者细胞学图像中观察到的变化表明了该药物的治疗效果。另一方面，在上皮细胞质中观察到的变化，如自噬（在CHO-K1细胞中明显促进）和白细胞吞噬现象可能表明布地奈德有额外的作用机制。
