Wang Ling-Feng, Tai Chih-Feng, Chien Chen-Yu, Chiang Feng-Yu, Chen Jeff Yi-Fu
Department of Otolaryngology, Kaohsiung Municipal Ta-Tung Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan; Department of Otolaryngology, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
Department of Otolaryngology, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan; Department of Otolaryngology, Kaohsiung Municipal Hsiao-Kang Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan.
Kaohsiung J Med Sci. 2015 May;31(5):235-40. doi: 10.1016/j.kjms.2015.02.001. Epub 2015 Mar 6.
Vitamin D and its derivatives have modulatory effects in immunological and inflammatory responses. Such properties suggest that they might have an impact on chronic inflammatory airway diseases, including nasal polyposis. The aim of this study was to understand the role of vitamin D in chronic rhinosinusitis with nasal polyps (CRSwNP) by investigating its effect on the secretion of matrix metalloproteinase-2 (MMP-2) and MMP-9 in nasal polyp-derived fibroblasts. Two primary fibroblast cultures were established from nasal polyp tissues obtained during surgery. The nasal polyp-derived fibroblasts were stimulated with tumor necrosis factor-α (TNF-α; 10 ng/mL) for 24 hours, followed by replacement with media alone or with vitamin D derivatives (calcitriol or tacalcitol; 10μM) and incubated for another 24 hours. After the treatments, the levels of MMP-2 and MMP-9 secreted were evaluated by both enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. ELISA results revealed that TNF-α could substantially stimulate the secretion of MMP-2 (p < 0.01) and MMP-9 (p < 0.001) in nasal polyp-derived fibroblasts. More importantly, such stimulatory effect was significantly suppressed by adding calcitriol (p ≤ 0.01 for MMP-2 and p < 0.001 for MMP-9) or tacalcitol (p < 0.005 for both MMP-2 and MMP-9). The ELISA results were also confirmed by Western blot analysis. The inhibitory effect of vitamin D derivatives on MMP-2 and MMP-9 secretion could potentiate their application in pharmacotherapy of Taiwanese CRSwNP patients.
维生素D及其衍生物在免疫和炎症反应中具有调节作用。这些特性表明它们可能对慢性炎症性气道疾病,包括鼻息肉,产生影响。本研究的目的是通过研究维生素D对鼻息肉来源的成纤维细胞中基质金属蛋白酶-2(MMP-2)和MMP-9分泌的影响,来了解维生素D在伴有鼻息肉的慢性鼻窦炎(CRSwNP)中的作用。从手术中获得的鼻息肉组织建立了两种原代成纤维细胞培养物。用肿瘤坏死因子-α(TNF-α;10 ng/mL)刺激鼻息肉来源的成纤维细胞24小时,然后换成单独的培养基或添加维生素D衍生物(骨化三醇或他卡西醇;10μM),再孵育24小时。处理后,通过酶联免疫吸附测定(ELISA)和蛋白质印迹分析评估分泌的MMP-2和MMP-9水平。ELISA结果显示,TNF-α可显著刺激鼻息肉来源的成纤维细胞中MMP-2(p < 0.01)和MMP-9(p < 0.001)的分泌。更重要的是,添加骨化三醇(MMP-2为p ≤ 0.01,MMP-9为p < 0.001)或他卡西醇(MMP-2和MMP-9均为p < 0.005)可显著抑制这种刺激作用。蛋白质印迹分析也证实了ELISA结果。维生素D衍生物对MMP-2和MMP-9分泌的抑制作用可能会增强它们在台湾CRSwNP患者药物治疗中的应用。