NLRP3 inflammasome activation results in hepatocyte pyroptosis, liver inflammation, and fibrosis in mice.
NLRP3 inflammasome activation results in hepatocyte pyroptosis, liver inflammation, and fibrosis in mice.
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DOI：
10.1002/hep.26592
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发表时间：
2014-03
期刊：
影响因子：
13.5
通讯作者：
Feldstein, Ariel E.
中科院分区：
文献类型：
--
作者：
Wree, Alexander;Eguchi, Akiko;McGeough, Matthew D.;Pena, Carla A.;Johnson, Casey D.;Canbay, Ali;Hoffman, Hal M.;Feldstein, Ariel E.
最近已经认识到炎症体激活在药物诱导的和肥胖相关的肝病的发展中起核心作用。然而，炎性小体介导的肝损伤的来源和机制仍然知之甚少。我们的目的是使用新的小鼠模型研究NLRP 3炎性小体激活对肝脏的影响。我们产生了表达D301 N Nlrp 3突变（人NLRP 3中D303 N的直系同源物）的全局和骨髓细胞特异性条件突变Nlrp 3敲入小鼠，导致组成性激活的NLRP 3。为了研究NLRP 3启动的细胞死亡的存在和意义，我们从非实质细胞中分离肝细胞，并开发了一种新的基于流式细胞术（FACS）的策略，以检测和定量体内细胞死亡，该策略基于检测活性半胱天冬酶1和碘化丙啶（PI）阳性细胞。通过流式细胞术和基因表达分析对肝脏炎症进行组织学定量。通过肝星状细胞（HSC）活化标志物的红染色和qPCR评估肝纤维化。NLRP 3激活导致生存期缩短、生长不良和严重肝脏炎症;其特征在于肝脏中的嗜中性粒细胞浸润和HSC激活以及胶原沉积。阿那白滞素（一种白细胞介素-1受体拮抗剂）治疗可部分减弱这些变化。值得注意的是，来自全局Nlrp 3突变小鼠的肝细胞显示出显著的肝细胞热变性细胞死亡，活性半胱天冬酶1-PI双阳性细胞增加超过5倍。髓样细胞限制性突变NLRP 3激活导致在不存在可检测到的肝细胞死亡的情况下不太严重的肝脏表型。我们的数据表明，全球和在较小程度上骨髓特异性NLRP 3炎性体激活导致严重的肝脏炎症和纤维化，同时确定肝细胞热变性细胞死亡作为NLRP 3介导的肝损伤的新机制。
Inflammasome activation has been recently recognized to play a central role in the development of drug-induced and obesity-associated liver disease. However, the sources and mechanisms of inflammasome mediated liver damage remain poorly understood. Our aim was to investigate the effect of NLRP3 inflammasome activation on the liver using novel mouse models. We generated global and myeloid cell specific conditional mutant Nlrp3 knock-in mice expressing the D301N Nlrp3 mutation (ortholog of D303N in human NLRP3) resulting in a constitutively activated NLRP3. To study the presence and significance of NLRP3 initiated pyroptotic cell death, we separated hepatocytes from non-parenchymal cells and developed a novel flow cytometry-based (FACS) strategy to detect and quantify pyroptosis in vivo based on detection of active caspase1 and propidium iodide (PI) positive cells. Liver inflammation was quantified histologically, by FACS and via gene expression analysis. Liver fibrosis was assessed by Sirius-Red-staining and qPCR for markers of hepatic stellate cell-(HSC)-activation. NLRP3 activation resulted in shortened survival, poor growth, and severe liver inflammation; characterized by neutrophilic infiltration and HSC-activation with collagen deposition in the liver. These changes were partially attenuated by treatment with anakinra, an interleukin-1 receptor antagonist. Notably, hepatocytes from global Nlrp3 mutant mice showed marked hepatocyte pyroptotic cell death with more than a fivefold increase in active caspase1-PI double positive cells. Myeloid cell restricted mutant NLRP3 activation resulted in a less severe liver phenotype in the absence of detectable pyroptotic hepatocyte cell death. Our data demonstrates that global and to a lesser extent myeloid-specific NLRP3 inflammasome activation results in severe liver inflammation and fibrosis, while identifying hepatocyte pyroptotic cell death as a novel mechanism of NLRP3 mediated liver damage.
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影响因子：
16
作者：
Martinon, F;Burns, K;Tschopp, J
通讯作者：
Tschopp, J
影响因子：
13.5
作者：
Csak, Timea;Ganz, Michal;Pespisa, Justin;Kodys, Karen;Dolganiuc, Angela;Szabo, Gyongyi
通讯作者：
Szabo, Gyongyi
影响因子：
25.7
作者：
Feldstein, AE;Canbay, A;Gores, GJ
通讯作者：
Gores, GJ
影响因子：
3.6
作者：
Brennan, MA;Cookson, BT
通讯作者：
Cookson, BT
影响因子：
3
作者：
Clausen, BE;Burkhardt, C;Förster, I
通讯作者：
Förster, I