Regulation of TNF-α by 1α,25-dihydroxyvitamin D3 in human macrophages from CAPD patients
Regulation of TNF-α by 1α,25-dihydroxyvitamin D3 in human macrophages from CAPD patients
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DOI：
10.1046/j.1523-1755.2001.00467.x
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发表时间：
2001-01-01
影响因子：
19.6
通讯作者：
Shany, S
中科院分区：
文献类型：
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作者：
Cohen, ML;Douvdevani, A;Shany, S
背景我们以前曾报道过1 α，25-二羟维生素D-3 [1 α，25（OH）（2）D-3]在接受持续非卧床腹膜透析（CAPD）治疗的尿毒症患者的透析液中蓄积。据报道，该代谢物调节单核细胞/巨噬细胞产生细胞因子。由于肿瘤坏死因子-α（TNF-α）在腹膜炎期间启动炎症级联反应，本研究的目的是研究1 α，25（OH）（2）D-3对人腹膜巨噬细胞（HPMs）产生TNF-α的影响。从CAPD患者中获得HPM。将细胞与不同浓度的1 α，25（OH）（2）D-3、1 α，24（S）二羟基维生素D-2 [1 α，24（S）（OH）（2）D-2]或25-羟基维生素D-3（25-OH-D-3）孵育16小时。随后进行脂多糖（LPS; 1 μ g/mL）孵育2.5至6小时。通过酶联免疫吸附测定法测定TNF-α蛋白的产生。采用逆转录-聚合酶链反应方法检测肿瘤坏死因子-α mRNA的表达，并使用内部合成mRNA标准品进行定量。在LPS刺激前用1 α，25（OH）（2）D-3孵育HPMs可剂量依赖性地抑制TNF-α在mRNA和蛋白水平上的表达。用较少钙的维生素D-2类似物1 α，24（S）（OH）（2）D-2获得了类似的结果。HPM与25-OH-D-3的孵育也显示TNF-α的下调。表情由于这种下调作用被酮康唑阻断，因此这种作用可能是由HPMs将25-OH-D-3转化为1 α，25（OH）（2）D-3引起的。1 α，25（OH）（2）D-3对LPS激活的HPMs产生TNF-α具有强效抑制作用。我们假设1 α，25（OH）（2）D-3可能构成一种调节机制，通过控制腹膜炎症反应的强度，将在腹膜炎期间减轻组织损伤。
Background We have previously reported that 1 alpha ,25-dihydroxyvitamin D-3 [1 alpha ,25(OH)(2)D-3] accumulates in the dialysis fluid of uremic patients treated by continuous ambulatory peritoneal dialysis (CAPD). It has been reported that this metabolite regulates the production of cytokines by monocytes/macrophages. Since tumor necrosis factor-alpha (TNF-alpha) initiates an inflammatory cascade during peritonitis, the aim of the present study was to investigate the effect of 1 alpha ,25(OH)(2)D-3 on the production of TNF-alpha by human peritoneal macrophages (HPMs).Methods. HPMs were obtained from patients on CAPD. Cells were incubated with various concentrations of 1 alpha ,25(OH)(2)D-3, 1 alpha ,24(S) dihydroxyvitamin D-2 [1 alpha ,24(S)(OH)(2)D-2] Or 25-hydroxyvitamin D-3 (25-OH-D-3) for 16 hours. This was followed by lipopolysaccharide (LPS; 1 mug/mL) incubation for 2.5 to 6 hours. TNF-alpha protein production was determined by enzyme-linked immunosorbent assay. TNF-alpha mRNA was assayed by the reverse transcriptase-polymerase chain reaction procedure, using internal synthetic mRNA standards for quantitative results.Results. Incubation of HPMs with 1 alpha ,25(OH)(2)D-3 prior to stimulation with LPS dose dependently inhibited the expression of TNF-alpha on both mRNA and protein levels. Similar results were obtained with the less calcemic vitamin D-2 analogue 1 alpha ,24(S)(OH)(2)D-2. Incubation of HPMs with 25-OH-D-3 also revealed a down-regulation of TNF-alpha. expression. Since this down-regulatory effect was blocked by ketoconazole, it is likely that this effect was caused by the conversion of 25-OH-D-3 into 1 alpha ,25(OH)(2)D-3 by HPMs.Conclusions. 1 alpha ,25(OH)(2)D-3 has a potent inhibitory effect on the production of TNF-alpha by LPS-activated HPMs. We hypothesize that 1 alpha ,25(OH)(2)D-3 may constitute a regulatory mechanism that, by controlling the intensity of the inflammatory response of the peritoneum, will moderate tissue damage during peritonitis.