Nuclear Receptor Subfamily 1 Group D Member 1 Regulates Circadian Activity of NLRP3 Inflammasome to Reduce the Severity of Fulminant Hepatitis in Mice.
Nuclear Receptor Subfamily 1 Group D Member 1 Regulates Circadian Activity of NLRP3 Inflammasome to Reduce the Severity of Fulminant Hepatitis in Mice.
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DOI：
10.1053/j.gastro.2017.12.019
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发表时间：
2018-04
期刊：
影响因子：
29.4
通讯作者：
Duez HM
中科院分区：
文献类型：
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作者：
Pourcet B;Zecchin M;Ferri L;Beauchamp J;Sitaula S;Billon C;Delhaye S;Vanhoutte J;Mayeuf-Louchart A;Thorel Q;Haas JT;Eeckhoute J;Dombrowicz D;Duhem C;Boulinguiez A;Lancel S;Sebti Y;Burris TP;Staels B;Duez HM
先天免疫系统不仅对细菌信号作出反应，还对组织损伤后激活NLRP3炎性体复合物的非感染性危险相关分子模式作出反应。免疫功能在一天中会发生变化，但尚不清楚这些变化是否会影响NLRP3炎性体的活性。我们研究了核心时钟组件核受体亚家族1组D成员1 （NR1D1，也称为Rev-erbα）是否调节NLRP3炎症小体的表达、活性及其信号通路。我们收集了Nr1d1−/−小鼠及其Nr1d1+/+幼崽（对照）的naïve腹腔巨噬细胞和血浆，分析了NLR家族pyrin结构域3 （NLRP3）、白细胞介素1 β （IL1B，血浆）和IL18（血浆）的表达。我们还从这些小鼠身上收集了骨髓来源的原代巨噬细胞。在人原代巨噬细胞中，用小发夹rna敲低NR1D1水平。用脂多糖（LPS）孵育小鼠骨髓源性原代巨噬细胞和人原代巨噬细胞，诱导NLRP3、IL1B和IL18的表达；将细胞与LPS和ATP孵育以激活NLRP3复合物。我们分析了caspase 1的活性和细胞因子的分泌。NR1D1在原代小鼠和人巨噬细胞中被SR9009激活；部分细胞也用NLRP3抑制剂或caspase 1抑制剂孵育。Nr1d1−/−小鼠和对照小鼠腹腔注射LPS诱导腹膜炎症；分离血浆样本，测定细胞因子水平。注射SR9009的Nr1d1−/−小鼠、对照小鼠和对照小鼠分别给予LPS和半乳糖胺诱导暴发性肝炎和MCC950特异性抑制NLRP3；收集血浆检测细胞因子和肝衰竭标志物（ALAT）；收集肝组织，采用定量PCR、免疫组织化学和流式细胞术进行分析。在腹膜巨噬细胞中，NLRP3的表达及其复合物的激活随着一天中的时间（昼夜节律）而变化-这种调节需要NR1D1。与表达Nr1d1的巨噬细胞相比，Nr1d1 - / -小鼠和人巨噬细胞的原代巨噬细胞NLRP3的表达模式发生了改变，IL1B和1L18的产生模式也发生了改变。Nr1d1被破坏的小鼠比对照组小鼠发生更严重的急性腹膜炎症和暴发性肝炎。用NR1D1激活剂SR9009孵育巨噬细胞可降低NLRP3的表达和细胞因子的分泌。给予SR9009的小鼠比给予生理盐水（对照组）的小鼠肝功能衰竭程度较轻，存活时间更长。在Nr1d1 - / -小鼠和人巨噬细胞的药理激活研究中，我们发现Nr1d1可以调节巨噬细胞NLRP3的表达时间和炎症细胞因子的产生。激活NR1D1可减轻小鼠腹膜炎症和暴发性肝炎的严重程度。
The innate immune system responds not only to bacterial signals, but also to non-infectious danger-associated molecular patterns that activate the NLRP3 inflammasome complex after tissue injury. Immune functions vary over the course of the day, but it is not clear whether these changes affect the activity of the NLRP3 inflammasome. We investigated whether the core clock component nuclear receptor subfamily 1 group D member 1 (NR1D1, also called Rev-erbα) regulates expression, activity of the NLRP3 inflammasome, and its signaling pathway. We collected naïve peritoneal macrophages and plasma, at multiple times of day, from Nr1d1−/− mice and their Nr1d1+/+ littermates (controls) and analyzed expression of NLR family pyrin domain containing 3 (NLRP3), interleukin 1 beta (IL1B, in plasma), and IL18 (in plasma). We also collected bone marrow-derived primary macrophages from these mice. Levels of NR1D1 were knocked down with small hairpin RNAs in human primary macrophages. Bone marrow-derived primary macrophages from mice and human primary macrophages were incubated with lipopolysaccharide (LPS) to induce expression of NLRP3, IL1B and IL18; cells were incubated with LPS and ATP to activate the NLRP3 complex. We analyzed caspase 1 activity and cytokine secretion. NR1D1 was activated in primary mouse and human macrophages by incubation with SR9009; some of the cells were also incubated with an NLRP3 inhibitor or inhibitors of caspase 1. Nr1d1−/− mice and control mice were given intraperitoneal injections of LPS to induce peritoneal inflammation; plasma samples were isolated and levels of cytokines were measured. Nr1d1−/− mice, control mice, and control mice given injections of SR9009 were given LPS and galactosamine to induce fulminant hepatitis and MCC950 to specifically inhibit NLRP3; plasma was collected to measure cytokines and a marker of liver failure (ALAT); liver tissues were collected and analyzed by quantitative PCR, immunohistochemistry, and flow cytometry. In peritoneal macrophages, expression of NLRP3 and activation of its complex varied with time of day (circadian rhythm)—this regulation required NR1D1. Primary macrophages from Nr1d1−/− mice and human macrophages with knockdown of NR1D1 had altered expression patterns of NLRP3, compared to macrophages that expressed NR1D1, and altered patterns of IL1B and 1L18 production. Mice with disruption of Nr1d1 developed more-severe acute peritoneal inflammation and fulminant hepatitis than control mice. Incubation of macrophage with the NR1D1 activator SR9009 reduced expression of NLRP3 and secretion of cytokines. Mice given SR9009 developed less-severe liver failure and had longer survival times than mice given saline (control). In studies of Nr1d1−/− mice and human macrophages with pharmacologic activation of NR1D1, we found NR1D1 to regulate the timing of NLRP3 expression and production of inflammatory cytokines by macrophages. Activation of NR1D1 reduced the severity of peritoneal inflammation and fulminant hepatitis in mice.
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影响因子：
82.9
作者：
通讯作者：
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影响因子：
64.8
作者：
Gerhart-Hines Z;Feng D;Emmett MJ;Everett LJ;Loro E;Briggs ER;Bugge A;Hou C;Ferrara C;Seale P;Pryma DA;Khurana TS;Lazar MA
通讯作者：
Lazar MA
影响因子：
16.6
作者：
Banerjee, Subhashis;Wang, Yongjun;Burris, Thomas P.
通讯作者：
Burris, Thomas P.
DOI：
10.1073/pnas.1106750109
发表时间：
2012-01-10
影响因子：
11.1
作者：
Gibbs, Julie E.;Blaikley, John;Loudon, Andrew S. I.
通讯作者：
Loudon, Andrew S. I.
影响因子：
64.8
作者：
通讯作者：
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