Calcium sensing receptor activation in THP-1 macrophages triggers NLRP3 inflammasome and human preadipose cell inflammation
Calcium sensing receptor activation in THP-1 macrophages triggers NLRP3 inflammasome and human preadipose cell inflammation
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DOI：
10.1016/j.mce.2019.110654
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发表时间：
2020-02-05
影响因子：
4.1
通讯作者：
Cifuentes, Mariana
中科院分区：
文献类型：
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作者：
D'Espessailles, Amanda;Santillana, Natalia;Cifuentes, Mariana
过量的脂肪组织（AT）与炎症和肥胖相关疾病有关。我们研究了THP-1巨噬细胞中钙敏感受体（CaSR）介导的NLRP 3炎性小体激活是否会升高LS 14前脂肪细胞中的炎症，模拟有害的AT细胞串扰。暴露于西那卡塞（CaSR激活剂，2 μ M，4小时）的THP-1巨噬细胞显示促炎标志物和NLRP 3炎性体mRNA、pro-IL-1 β蛋白和caspase-1活性升高，而与CaSR负调节剂预孵育可防止这些作用。在THP-1细胞中沉默关键的NLRP 3炎性体组分ASC（siRNA），并评价炎性体活化（qPCR、蛋白质印迹、半胱天冬酶-1活性）或进一步培养它们以获得条件培养基（CoM）。将LS 14前脂肪细胞暴露于来自西那卡塞处理的THP-1的CoM升高了LS 14促炎细胞因子的表达，其被THP-1炎性体沉默废除。因此，CaSR活化通过巨噬细胞NLRP 3炎性体活化提高了LS 14前脂肪细胞中THP-1诱导的炎症。调节CaSR活化可以防止AT中有害的促炎细胞串扰，这是肥胖相关代谢紊乱的一种有前途的方法。
Excess adipose tissue (AT) associates with inflammation and obesity-related diseases. We studied whether calcium-sensing receptor (CaSR)-mediated NLRP3 inflammasome activation in THP-1 macrophages elevates inflammation in LS14 preadipocytes, modeling deleterious AT cell crosstalk. THP-1 macrophages exposed to cinacalcet (CaSR activator, 2 mu M, 4 h) showed elevated proinflammatory marker and NLRP3 inflammasome mRNA, pro-IL-1 beta protein and caspase-1 activity, whereas preincubation with CaSR negative modulators prevented these effects. The key NLRP3 inflammasome component ASC was silenced (siRNA) in THP-1 cells, and inflammasome activation was evaluated (qPCR, Western blot, caspase-1 activity) or they were further cultured to obtain conditioned medium (CoM). Exposure of LS14 preadipocytes to CoM from cinacalcet-treated THP-1 elevated LS14 proinflammatory cytokine expression, which was abrogated by THP-1 inflammasome silencing. Thus, CaSR activation elevates THP-1-induced inflammation in LS14 preadipocytes, via macrophage NLRP3 inflammasome activation. Modulating CaSR activation may prevent deleterious proinflammatory cell crosstalk in AT, a promising approach in obesity-related metabolic disorders.