ADAM10 modulates calcitriol-regulated RAGE in cardiomyocytes
ADAM10 modulates calcitriol-regulated RAGE in cardiomyocytes
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DOI：
10.1111/eci.12789
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发表时间：
2017-09-01
影响因子：
5.5
通讯作者：
Chen, Yi-Jen
中科院分区：
文献类型：
--
作者：
Lee, Ting-Wei;Kao, Yu-Hsun;Chen, Yi-Jen
晚期糖基化终产物受体（RAGE）信号传导在心血管疾病的发病机制中起着关键作用。骨化三醇调节心脏RAGE表达。本研究探讨了骨化三醇对心肌细胞RAGE和可溶性RAGE （sRAGE）表达影响的机制。材料和方法采用western blot、ELISA、荧光定量分析和PCR分析，分别检测在不含骨化三醇（10和100 nM）、核因子- b （NF-B）抑制剂（50 g/mL）和不含骨化三醇的HL-1心房肌细胞中RAGE、sRAGE、内源性分泌RAGE （esRAGE）、Jun n -末端激酶（JNK）、崩解素和金属蛋白酶10 （ADAM10）的表达和酶活性。结果与对照细胞相比，scalcitriol （10 nM）显著降低了HL-1心肌细胞中RAGE蛋白的表达，增加了sRAGE浓度。这些变化与ADAM10蛋白表达和酶活性升高以及esRAGE mRNA表达升高有关。然而，在ADAM10抑制剂存在时，骨化三醇对RAGE的抑制作用减弱。甲基乙二醛（500 M, 10 min）介导的JNK磷酸化在骨化三醇（10 nM）存在下减弱。此外，对照组和NF-B抑制剂处理的HL-1细胞具有相似的RAGE和sRAGE表达，这表明骨化三醇介导的RAGE调节不依赖于NF-B信号传导。结论骨化三醇通过激活心肌细胞ADAM10介导RAGE下调和sRAGE生成增加。结果提示骨化三醇在治疗rage介导的心血管并发症方面具有治疗潜力。
BackgroundReceptor for advanced glycation end products (RAGE) signalling plays a critical role in the pathogenesis of cardiovascular disease. Calcitriol modulates cardiac RAGE expression. This study explored the mechanisms underlying the effect of calcitriol on RAGE and soluble RAGE (sRAGE) expression in cardiomyocytes.Materials and methodsWestern blot, ELISA, fluorometric assay and PCR analyses were used to evaluate the RAGE, sRAGE, endogenous secretory RAGE (esRAGE), Jun N-terminal kinase (JNK), and a disintegrin and metalloprotease 10 (ADAM10) expression and enzyme activity in HL-1 atrial myocytes without and with calcitriol (10 and 100 nM), nuclear factor-B (NF-B) inhibitor (50 g/mL), or ADAM10 inhibitor (5 M) incubation for 48 h.ResultsCalcitriol (10 nM) significantly reduced RAGE protein expression and increased sRAGE concentrations in HL-1 cardiomyocytes compared with control cells. These changes were associated with increased protein expression and enzyme activity of ADAM10 and higher mRNA expression of esRAGE. In the presence of ADAM10 inhibitor, however, the suppressive effect of calcitriol on RAGE was diminished. Methylglyoxal (500 M for 10 min)-mediated JNK phosphorylation was attenuated in the presence of calcitriol (10 nM). Moreover, control and NF-B inhibitor-treated HL-1 cells had similar RAGE and sRAGE expression, suggesting that calcitriol-mediated RAGE modulation was independent of NF-B signalling.ConclusionsWe showed that RAGE downregulation and increased sRAGE production by calcitriol were mediated through ADAM10 activation in cardiomyocytes. The results suggest that calcitriol has therapeutic potential in treating RAGE-mediated cardiovascular complications.