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中华临床医师杂志(电子版) ›› 2022, Vol. 16 ›› Issue (05) : 447 -451. doi: 10.3877/cma.j.issn.1674-0785.2022.05.014

基础研究

盐皮质受体对脂多糖诱导的巨噬细胞NOD样受体热蛋白结构域相关蛋白3炎症复合体激活的作用及其机制
苏程程1, 马永强2, 郎胜坤3, 刘斌1, 魏路清1,†(), 姬文婕2,†()   
  1. 1. 300162　天津，武警特色医学中心呼吸与危重症医学科
    2. 300162　天津，武警特色医学中心心内科
    3. 300162　天津，武警特色医学中心医学影像科
  • 收稿日期:2022-03-11 出版日期:2022-05-15
  • 通信作者: 魏路清, 姬文婕
  • 基金资助:
    天津市自然科学基金(18JCQNJC12000)

Role and mechanism of action of mineralocorticoid receptorin lipopolysaccharide induced NLRP3 inflammasomes activation

Chengcheng Su1, Yongqiang Ma2, Shengkun Lang3, Bin Liu1, Luqing Wei1,†(), Wenjie Ji2,†()   

  1. 1. Department of Respiratory and Critical Care Medicine, Characteristic Medical Center of Chinese People's Armed Police Force, Tianjin 300162, China
    2. Institutes of Cardiovascular Disease and Heart Center, Characteristic Medical Center of Chinese People's Armed Police Force, Tianjin 300162, China
    3. Department of Radiology, Characteristic Medical Center of Chinese People's Armed Police Force, Tianjin 300162, China
  • Received:2022-03-11 Published:2022-05-15
  • Corresponding author: Luqing Wei, Wenjie Ji
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引用本文：

苏程程, 马永强, 郎胜坤, 刘斌, 魏路清, 姬文婕. 盐皮质受体对脂多糖诱导的巨噬细胞NOD样受体热蛋白结构域相关蛋白3炎症复合体激活的作用及其机制[J]. 中华临床医师杂志(电子版), 2022, 16(05): 447-451.

Chengcheng Su, Yongqiang Ma, Shengkun Lang, Bin Liu, Luqing Wei, Wenjie Ji. Role and mechanism of action of mineralocorticoid receptorin lipopolysaccharide induced NLRP3 inflammasomes activation[J]. Chinese Journal of Clinicians(Electronic Edition), 2022, 16(05): 447-451.

目的

探讨盐皮质受体对脂多糖（LPS）诱导巨噬细胞炎症复合体激活的作用，并探讨其机制。

方法

对小鼠巨噬细胞系RAW264.7，用LPS、嘌呤受体激动剂腺苷5′-（3-硫代三磷酸盐）四锂盐（ATP-γ-s）、盐皮质激素醛固酮（Ald）干预建立NOD样受体热蛋白结构域相关蛋白3（NLRP3）炎症复合体模型；运用盐皮质受体阻滞剂螺内酯（SPI）、P2X7嘌呤受体拮抗剂（A438）对LPS诱导的炎症复合体模型进行干预，分为生理盐水（NS）组、LPS组、LPS+SPI组、LPS+A438组、LPS+SPI+A438组，实时定量聚合酶链反应（RT-PCR）法检测各组NLRP3炎症复合体的基因表达水平；使用三磷酸腺苷（ATP）检测试剂盒，检测LPS、Ald作用于RAW264.7细胞后培养上清ATP水平，并检测SPI对上述过程的干预作用。

结果

LPS、Ald、ATP-γ-s干预RAW264.7细胞，细胞NLRP3的基因表达水平分别较NS组上调（2.51±0.42）、（2.22±0.28）、（2.07±0.11）倍，差异均有统计学意义（均P＜0.05）；SPI、A438单独干预或SPI+A438同时干预，可以下调NLRP3的表达至NS组的（1.39±0.20）、（1.31±0.15）、（1.25±0.09）倍，均较LPS组显著下降（均P＜0.05）。LPS、Ald干预RAW264.7显著提高了细胞上清ATP水平，而SPI干预显著降低了LPS、Ald干预所致ATP释放，差异均有统计学意义（均P＜0.05）。

结论

SPI可能是通过阻断盐皮质受体以降低胞外ATP的释放，从而抑制LPS导致的NLRP3炎症复合体的激活。

关键词: 盐皮质激素, 脂多糖, 螺内酯, NOD样受体热蛋白结构域相关蛋白3, 三磷酸腺苷
Objective

To investigate the influence of spironolactone on lipopolysaccharide-induced activation of nucleotide-binding oligomerization domain NOD-like receptor 3 (NLRP3) in macrophages and explore the exact mechanism involved.

Methods

The mousemacrophage cellline RAW264.7 was treated with lipopolysaccharide (LPS), aldosterone (Ald), and adenosine 5'-O-3-thiotriphosphate (ATP-γ-s) to establish anNLRP3activation model. Then,themineralocorticoid receptor blocker spironolactone(SPI) and P2X7 receptor antagonist A438, alone or in combination, were used to intervene the NLRP3 activation model,and the NLRP3 expression level inRAW264.7 cells was detected using real-time polymerase chain reaction (RT-PCR). The supernatant triphosadenine(ATP) concentrationsin RAW264.7 cells challenged withLPS, Ald, LPS+SPI, and Ald+SPI were tested.

Results

LPS,Ald,and ATP-γ-s can activate NLRP3 inflammasomes, and the NLRP3 gene expression levels were significantly elevated by (2.51±0.42), (2.22±0.28), and (2.07±0.11) times, respectively, compared with that inthe NS group (P＜0.05). The NLRP3 activation level in the LPS groupwassignificantly down-regulate by SPI, A438, SPI+A438 by (1.39±0.20), (1.31±0.15), and (1.25±0.09) times, respectively, compared with that inthe NS group (P＜0.05). The supernatant ATP concentration inRAW264.7 cells challenged by LPS and Ald waselevated significantly when compared with that in theNS group (P＜0.05), while SPI treatment can reduce ATP concentration significantly in LPS and Ald-challenged cell supernatant (P＜0.05).

Conclusion

Blocking mineralocorticoid receptor using SPI can devitalize NLRP3 activation viaextracellular ATP down-regulation.

Key words: Mineralocorticoid, Lipopolysaccharide, Spironolactone, Nucleotide-binding oligomerization domain NOD-like receptor 3, Triphosadenine
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