Rostkowska-Nadolska Beata, Fraczek Marcin, Gawron Wojciech, Latocha Małgorzata
Department of Otolaryngology Head & Neck Surgery, Wroclaw Medical University, Wrocław, Poland.
Acta Biochim Pol. 2009;56(2):235-42. Epub 2009 Jun 6.
Vitamin D (VD) and its different analogues, besides their classic role as regulators of calcium and phosphor homeostasis, have emerged as a large family of antiproliferative agents. Such properties suggested VD potential as a therapy for chronic inflammatory diseases, including nasal polyposis (NP). NP growth involves both an inflammatory process and the proliferation of fibroblast as an important factor inducing aberrations in the phenotype of the epithelium. The aim of this study was to investigate the possible influence of 1alpha,25-dihydroxyvitamin D(3) (calcitriol) and 1alpha,24(R)-dihydroxyvitamin D(3) (tacalcitol) in monotherapy and in combination with budesonid R (BR) on NP fibroblast proliferation.
The study involved 26 samples of NP. NP cells were cultured on 96-well plates beginning with a concentration of 5 x 10(3) cells per well with RPMI 1640 medium supplemented with antibiotics and 10% foetal bovine serum. After the fourth to sixth passage the medium was replaced with a nutrient medium with calcitriol or tacalcitol in a defined concentration (from 10(-9) M to 10(-3) M) alone or in combination with BR in 1:1, 1:3 or 3:1 ratios, each at concentrations from 10(-5) M to 10(-3) M.
Growth inhibition of nasal fibroblasts exposed to calcitriol or tacalcitol was noted. Significant antiproliferating activity was observed at calcitriol concentrations of 10(-4) M and 10(-3) M after 48 h, and at a concentration of 10(-3) M after 72 h with the percentage of proliferating cells reduced to 30% compared to the control samples (P < 0.05). In cells treated with tacalcitol the maximal effect was seen at 10(-4) M after 48 h and at 10(-3)M after 72 h with a 60% inhibition with respect to the control (P < 0.05). The inhibition of fibroblast proliferation reached the maximal level when they were exposed to calcitriol: BR (1 : 1) or tacalcitol: BR (1 : 1), each at a concentration of 10(-4) M, after 72 h (82% and 69%, respectively).
The antiproliferative activity of calcitriol and tacalcitol in NP cultures was confirmed. Because of its lower toxicity and higher activity tacalcitol seems to be the more promising agent in NP therapy, both as a single medication and in treatment protocols with BR.
维生素D（VD）及其不同类似物，除了作为钙和磷稳态调节剂的经典作用外，已成为一大类抗增殖剂。这些特性表明VD有可能用于治疗慢性炎症性疾病，包括鼻息肉（NP）。NP的生长涉及炎症过程和成纤维细胞的增殖，而成纤维细胞增殖是诱导上皮细胞表型异常的重要因素。本研究的目的是探讨1α,25 - 二羟基维生素D3（骨化三醇）和1α,24（R） - 二羟基维生素D3（他卡西醇）单药治疗以及与布地奈德R（BR）联合使用对NP成纤维细胞增殖的可能影响。
本研究涉及26份NP样本。NP细胞接种于96孔板，起始浓度为每孔5×10³个细胞，使用补充有抗生素和10%胎牛血清的RPMI 1640培养基。在传代4至6次后，将培养基替换为含有特定浓度（从10⁻⁹M至10⁻³M）的骨化三醇或他卡西醇的营养培养基，单独使用或与BR按1:1、1:3或3:1的比例联合使用，每种组合的浓度均为10⁻⁵M至10⁻³M。
观察到暴露于骨化三醇或他卡西醇的鼻成纤维细胞生长受到抑制。48小时后，骨化三醇浓度为10⁻⁴M和10⁻³M时观察到显著的抗增殖活性，72小时后浓度为10⁻³M时，增殖细胞百分比与对照样本相比降至30%（P < 0.05）。在用他卡西醇处理的细胞中，48小时后在10⁻⁴M时观察到最大效应，72小时后在10⁻³M时观察到最大效应，相对于对照有60%的抑制率（P < 0.05）。当成纤维细胞暴露于骨化三醇：BR（1:1）或他卡西醇：BR（1:1），每种浓度为10⁻⁴M，72小时后，成纤维细胞增殖的抑制达到最大水平（分别为82%和69%）。
证实了骨化三醇和他卡西醇在NP培养物中的抗增殖活性。由于其较低的毒性和较高的活性，他卡西醇似乎在NP治疗中更有前景，无论是作为单一药物还是与BR联合使用的治疗方案中。
