1α,25-dihydroxy-vitamin D3 stimulation of bronchial smooth muscle cells induces autocrine, contractility, and remodeling processes
1α,25-dihydroxy-vitamin D3 stimulation of bronchial smooth muscle cells induces autocrine, contractility, and remodeling processes
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DOI：
10.1152/physiolgenomics.00134.2006
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发表时间：
2007-04-24
影响因子：
4.6
通讯作者：
Hudson, Thomas J.
中科院分区：
文献类型：
--
作者：
Bosse, Yohan;Maghni, Karim;Hudson, Thomas J.
维生素D受体（VDR）基因的遗传变异最近与哮喘有关。解释这种关联的生物学机制尚不清楚，但考虑到其配体1 α，25-二羟基维生素D3 [1 α，25（OH）（2）D-3]的多效性效应，可能涉及许多细胞类型。鉴于支气管平滑肌细胞（BSMCs）在哮喘发病中的重要作用，本实验旨在探讨1 α，25（OH）（2）D-3对BSMCs基因表达的调控作用。RT-PCR和Western blot结果显示VDR在人BSMCs中表达于mRNA水平和蛋白水平。然后通过显示1 α，25（OH）（2）D-3刺激后24-羟化酶（CYP 24 A1）基因表达的> 200倍变化来证明受体的功能性。然后进行微阵列实验，以鉴定用1 α，25（OH）（2）D-3处理或未处理的BMSC中差异调节的基因和途径。使用稳健多芯片平均强度，U133 plus 2.0 Affytechnic GeneChip上总共729个探针组显示倍数变化差异高于1.5阈值。这对应于在1 α，25（OH）（2）D-3刺激后上调的231个独特基因和下调的215个独特基因。13个随机基因的芯片和实时荧光PCR结果之间具有较高的相似性，一致性相关系数为0.91。实时荧光定量PCR检测哮喘候选基因的表达。为了确定该调节的生物学相关性，进行了生物学途径分析。最重要的上调基因网络包括参与形态发生、细胞生长和存活的基因以及编码结构蛋白的基因。
Genetic variants in the vitamin D receptor (VDR) gene were recently associated with asthma. The biological mechanisms explaining this association are unknown but are likely to involve many cell types given the pleiotropic effect of its ligand, 1 alpha,25-dihydroxy-vitamin D3 [1 alpha,25(OH)(2)D-3]. Considering the prominent role of bronchial smooth muscle cells (BSMCs) in the pathogenesis of asthma, experiments were conducted to explore the gene regulatory effects of 1 alpha,25(OH)(2)D-3 in these cells. Using RT-PCR and Western blot, we showed that VDR is present both at the mRNA transcript and protein levels in human BSMCs. The functionality of the receptor was then demonstrated by showing a > 200-fold change in the expression of the 24-hydroxylase (CYP24A1) gene following 1 alpha,25(OH)(2)D-3 stimulation. Microarray experiments were then performed to identify differentially regulated genes and pathways in BMSCs treated or not with 1 alpha,25(OH)(2)D-3. A total of 729 probe sets on the U133 plus 2.0 Affymetrix GeneChip showed fold-change differences above the 1.5 threshold using the Robust Multichip Average intensities. This corresponds to 231 unique genes that were upregulated and 215 unique genes that were down-regulated following 1 alpha,25(OH)(2)D-3 stimulation. A high similarity between microarray and real-time PCR results was observed for 13 random genes, with a concordance correlation coefficient of 0.91. Real-time PCR was also performed to confirm the regulation of asthma candidate genes. To identify the biological relevance of this regulation, biological pathways analyses were performed. The most significant network of upregulated genes included genes involved in morphogenesis, cell growth, and survival as well as genes encoding structural