Production of 22-Hydroxy Metabolites of Vitamin D3 by Cytochrome P450scc (CYP11A1) and Analysis of Their Biological Activities on Skin Cells
Production of 22-Hydroxy Metabolites of Vitamin D3 by Cytochrome P450scc (CYP11A1) and Analysis of Their Biological Activities on Skin Cells
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DOI：
10.1124/dmd.111.040071
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发表时间：
2011-09-01
影响因子：
3.9
通讯作者：
Slominski, Andrzej
中科院分区：
文献类型：
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作者：
Tuckey, Robert C.;Li, Wei;Slominski, Andrzej
细胞色素P450SCC（CYP11A1）可将维生素D-3羟基化，产生20 S-羟基维生素D-3 [20（OH）D-3]和20 S，23-二羟基维生素D-3 [20，23（OH）（2）D-3]作为主要代谢产物。它们具有生物活性，可作为部分维生素D受体（VDR）激动剂。次要产品包括17-羟基维生素D-3、17，20-二羟基维生素D-3和17，20，23-三羟基维生素D-3。在本研究中，我们进一步分析了细胞色素P450SCC（P450SCC）作用于维生素D-3的反应产物，并鉴定了两种22-羟基衍生物作为产物，22-羟基维生素D-3 [22（OH）D-3]和20S，22-二羟基维生素D-3 [20，22（OH）（2）D-3]。这两种衍生物的结构均由NMR确定。P450SCC可将纯化的22（OH）D-3转化为20，22（OH）（2）D-3。20，22（OH）（2）D-3也可由20（OH）D3产生，并代谢为三羟基维生素D-3产物。我们比较了这些新衍生物与20（OH）D-3、20，23（OH）（2）D-3和1 α，25-二羟基维生素D-3 [1，25（OH）（2）D-3]的生物活性。1，25（OH）（2）D-3、20（OH）D-3、22（OH）D-3、20，23（OH）（2）D-3和20，22（OH）（2）D-3以剂量依赖性方式显著抑制角质形成细胞增殖。外皮蛋白表达（角质形成细胞分化的标志物）的最强诱导剂是20，23（OH）（2）D-3、20，22（OH）（2）D-3、20（OH）D-3和1，25（OH）（2）D-3，其中22（OH）D-3具有异质效应。除1，25（OH）（2）D-3外，所有检测的类似物均观察到对CYP 24 mRNA表达的刺激很小或没有刺激。所有化合物均能促进VDR从细胞质向细胞核的移位，其中22（OH）D-3和20，22（OH）（2）D-3的作用小于1，25（OH）（2）D-3和20（OH）D-3。因此，我们已经确定22（OH）D-3和20，22（OH）（2）D-3是CYP 11 A1对维生素D-3作用的产物，并表明与20（OH）D-3和20，23（OH）（2）D-3一样，它们通过VDR对角质形成细胞具有活性，然而，与其他P450 SCC衍生的维生素D-3羟基代谢物相比，表现出一定程度的表型异质性。
Cytochrome P450scc (CYP11A1) can hydroxylate vitamin D-3, producing 20S-hydroxyvitamin D-3 [20(OH)D-3] and 20S,23-dihydroxyvitamin D-3 [20,23(OH)(2)D-3] as the major metabolites. These are biologically active, acting as partial vitamin D receptor (VDR) agonists. Minor products include 17-hydroxyvitamin D-3, 17,20-dihydroxyvitamin D-3, and 17,20,23-trihydroxyvitamin D-3. In the current study, we have further analyzed the reaction products from cytochrome P450scc (P450scc) action on vitamin D-3 and have identified two 22-hydroxy derivatives as products, 22-hydroxyvitamin D-3 [22(OH)D-3] and 20S, 22-dihydroxyvitamin D-3 [20,22(OH)(2)D-3]. The structures of both of these derivatives were determined by NMR. P450scc could convert purified 22(OH)D-3 to 20,22(OH)(2)D-3. The 20,22(OH)(2)D-3 could also be produced from 20(OH) D3 and was metabolized to a trihydroxyvitamin D-3 product. We compared the biological activities of these new derivatives with those of 20(OH)D-3, 20,23(OH)(2)D-3, and 1 alpha,25-dihydroxyvitamin D-3 [1,25(OH)(2)D-3]. 1,25(OH)(2)D-3, 20(OH)D-3, 22(OH)D-3, 20,23(OH)(2)D-3, and 20,22(OH)(2)D-3 significantly inhibited keratinocyte proliferation in a dose-dependent manner. The strongest inducers of involucrin expression (a marker of keratinocyte differentiation) were 20,23(OH)(2)D-3, 20,22(OH)(2)D-3, 20(OH)D-3, and 1,25(OH)(2)D-3, with 22(OH)D-3 having a heterogeneous effect. Little or no stimulation of CYP24 mRNA expression was observed for all the analogs tested except for 1,25(OH)(2)D-3. All the compounds stimulated VDR translocation from the cytoplasm to the nucleus with 22(OH)D-3 and 20,22(OH)(2)D-3 having less effect than 1,25(OH)(2)D-3 and 20(OH)D-3. Thus, we have identified 22(OH)D-3 and 20,22(OH)(2)D-3 as products of CYP11A1 action on vitamin D-3 and shown that, like 20(OH)D-3 and 20,23(OH)(2)D-3, they are active on keratinocytes via the VDR, however, showing a degree of phenotypic heterogeneity in comparison with other P450scc-derived hydroxy metabolites of vitamin D-3.