Calcitriol blunts pro-atherosclerotic parameters through NFκB and p38 in vitro
Calcitriol blunts pro-atherosclerotic parameters through NFκB and p38 in vitro
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DOI：
10.1111/j.1365-2362.2008.01977.x
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发表时间：
2008-08-01
影响因子：
5.5
通讯作者：
Rashid, G.
中科院分区：
文献类型：
--
作者：
Talmor, Y.;Bernheim, J.;Rashid, G.
背景：维生素D-3代谢紊乱与心血管发病率和死亡率增加有关。本研究的目的是评估骨化三醇（维生素D-3的活性代谢物）对人脐静脉内皮细胞（HUVEC）促动脉粥样硬化参数的影响。材料与方法将10（-10）和/或10(-9)mol L-1的骨化三醇分别给予未刺激或脂多糖（LPS）刺激的培养HUVEC。流式细胞术检测细胞间黏附分子-1和血小板内皮细胞黏附分子-1。RT-PCR检测晚期糖基化终产物受体（RAGE）和白细胞介素-6 (IL-6) mRNA的表达，酶联免疫吸附试验（ELISA）检测IL-6的分泌。转录因子检测试剂盒检测核p65 dna结合活性，Western blot检测抑制因子κ B α (I κ B α)、磷酸化因子κ B α (P-I κ B α)和磷酸化因子p38有丝分裂原活化蛋白激酶（MAPK）蛋白水平。结果骨化三醇降低了粘附分子的表达，降低了脂多糖诱导的RAGE和IL-6 mRNA的表达，诱导了IL-6的分泌。此外，骨化三醇还降低了脂多糖诱导的核因子κ B (NF κ B)-p65 dna结合活性。骨化三醇处理后I κ B α水平升高，p-I κ B α水平降低。LPS处理后激活的p38 MAPK水平的增加也由于骨化三醇的预孵育而降低。结论骨化三醇治疗后NF κ B和p38活性的降低可能解释了骨化三醇的抗炎/动脉粥样硬化特性，证明了骨化三醇对促炎参数：粘附分子、RAGE和IL-6的抑制作用。
Background Disturbances in vitamin D-3 metabolism are associated with an increased cardiovascular morbidity and mortality. The aim of this study was to assess the effects of calcitriol, the active metabolite of vitamin D-3, on pro-atherosclerotic parameters in human umbilical vein cord endothelial cells (HUVEC).Materials and methods Calcitriol at 10(-10) and/or 10(-9) mol L-1 was given to cultured HUVEC which were either non-stimulated or lipopolysaccharide (LPS) stimulated. Inter cellular adhesion molecule-1 and platelet-endothelial cell adhesion molecule-1, were determined by flow cytometry analysis. The receptor of advanced glycation end product (RAGE) and interleukin-6 (IL-6) mRNA expressions by RT-PCR and IL-6 secretion by enzyme-linked immunosorbent assay (ELISA). Nuclear p65 DNA-binding activity was measured by transcription factor assay kit and the inhibitor-kappa B alpha (I kappa B alpha), phosphorylated-I kappa B alpha (P-I kappa B alpha) and phosphorylated-p38 mitogen-activated protein kinase (MAPK) protein levels were determined by Western blot.Results Calcitriol decreased the adhesion molecules expression, as well as the LPS-induced mRNA expressions of RAGE and IL-6 and LPS induced IL-6 secretion. Furthermore, the LPS induced nuclear factor kappa B (NF kappa B)-p65 DNA-binding activity was also decreased by calcitriol. I kappa B alpha levels were increased and p-I kappa B alpha levels decreased after calcitriol treatment. The increased levels of activated p38 MAPK after LPS treatment were also decreased due to pre-incubation with calcitriol.Conclusions The decreased NF kappa B and p38 activities followed by calcitriol treatment may explain the anti-inflammatory/atherosclerotic properties of calcitriol that were observed previously and were emphasized in this study, demonstrating the inhibitory effect of calcitriol on the pro-inflammatory parameters: adhesion molecules, RAGE and IL-6.