NLRP3 Deletion Inhibits the Non-alcoholic Steatohepatitis Development and Inflammation in Kupffer Cells Induced by Palmitic Acid
NLRP3 Deletion Inhibits the Non-alcoholic Steatohepatitis Development and Inflammation in Kupffer Cells Induced by Palmitic Acid
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NLRP3 缺失抑制棕榈酸诱导的非酒精性脂肪性肝炎的发展和库普弗细胞炎症
DOI：
10.1007/s10753-017-0628-z
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发表时间：
2017-12-01
期刊：
影响因子：
5.1
通讯作者：
He, Kun
中科院分区：
文献类型：
--
作者：
Cai, Can;Zhu, Xiwen;He, Kun
促炎细胞因子IL-1 β和IL-18的裂解和分泌受NLRP 3（含NACHT、LRR和PYD结构域的蛋白3）炎性体活化的调节。枯否细胞（KCs）参与各种肝脏疾病的发病机制，例如非酒精性脂肪性肝病（NAFLD）、酒精性肝病和肝纤维化。然而，NLRP 3在非酒精性脂肪性肝炎（NASH）中的作用尚未得到评估。本研究采用蛋氨酸胆碱缺乏（MCD）饲料建立小鼠NASH模型。检测肝组织中F4/80和NLRP 3的表达水平，并检测血清中IL-1 β和IL-18的水平。从野生型（WT）小鼠和NLRP 3敲除（NLRP 3（-/-））小鼠中分离KC，然后随机分为两组：对照组和棕榈酸（PA）组。RT-PCR、免疫印迹和免疫荧光法检测KC中NLRP 3、ASC和caspase-1的表达水平。采用酶联免疫吸附试验（ELISA）检测KCs培养上清中IL-1 β和IL-18的水平。我们发现KCs和NLRP 3在NASH的进展中发挥促炎作用，可能是通过PA诱导KCs分泌IL-1 β和IL-18。PA可作为一种损伤相关的分子模式，上调WT小鼠KC中NLRP 3、ASC和caspase-1的mRNA和蛋白表达水平。相反，NLRP 3缺失可抑制PA诱导的KC中NLRP 3炎性体的上调和活化。此外，在PA刺激下，WT小鼠KC SN中促炎细胞因子IL-1 β和IL-18的水平均升高，并且这些细胞因子在SN中的增加被NLRP 3缺失阻断。总之，我们的新发现表明NLRP 3在PA刺激诱导的NASH发展和促炎细胞因子IL-1 β和IL-18分泌中起关键作用，NLRP 3可能是治疗与NLRP 3炎性体激活相关的肝脏炎性疾病的有效潜在靶点。
The cleavage and secretion of pro-inflammatory cytokines IL-1 beta and IL-18 is regulated by NLRP3 (NACHT, LRR, and PYD domain-containing protein 3) inflammasome activation. Kupffer cells (KCs) are implicated in the pathogenesis of various liver diseases, such as non-alcoholic fatty liver disease (NAFLD), alcoholic liver disease, and liver fibrosis. However, the role of NLRP3 played in the non-alcoholic steatohepatitis (NASH) has yet to be evaluated. In the present study, methionine-choline-deficient (MCD) diet was used to establish the mice NASH model. The expression levels of F4/80 and NLRP3 in liver tissues were evaluated, and the IL-1 beta and IL-18 in serum were also evaluated. KCs were isolated from wild-type (WT) mice and NLRP3 knockout (NLRP3(-/-)) mice and then randomly divided into two groups: the control and palmitic acid (PA) groups. The expression levels of NLRP3, ASC, and caspase-1 in KCs were determined by RT-PCR, western blotting, and immunofluorescence. The levels of IL-1 beta and IL-18 in the supernatant (SN) of KCs were evaluated by enzyme-linked immunosorbent assay (ELISA). We found that KCs and NLRP3 play pro-inflammatory roles in the progression of NASH, probably through secretions of IL-1 beta and IL-18 by KCs induced by PA. PA could act as a kind of damage-associated molecular patterns to elevate the messenger RNA and protein expression levels of NLRP3, ASC, and caspase-1 in KCs from WT mice. In the contrast, NLRP3 deletion could inhibit the NLRP3 inflammasome upregulation and activation in KCs induced by PA. Furthermore, the levels of pro-inflammatory cytokines IL-1 beta and IL-18 in the SN of KCs from WT mice were all elevated with the stimulation of PA, and the increase of these cytokines in the SN was blocked by NLRP3 deletion. In conclusion, our novel findings demonstrate that NLRP3 plays a pivotal role in NASH development and pro-inflammatory cytokines IL-1 beta and IL-18 secretion induced by PA stimulation, and NLRP3 might be an effective potential target for the treatment of liver inflammatory diseases associated with NLRP3 inflammasome activation.