Zhang Yingyu, Jiang Yuanyuan, Zhang Daimo, Hu Xinyue, Deng Shuanglinzi, Li Xiaozhao, Feng Juntao
Department of Respiratory Medicine, National Key Clinical Specialty, Branch of National Clinical Research Center for Respiratory Disease, Xiangya Hospital, Central South University, Changsha, Hunan 410008, China.
Department of Clinical Laboratory, Xiangya Hospital, Central South University, Changsha, Hunan 410008, China.
Chin Med J Pulm Crit Care Med. 2024 Dec 17;2(4):279-288. doi: 10.1016/j.pccm.2024.11.007. eCollection 2024 Dec.
Glucocorticoid-induced transcript 1 (GLCCI1) has been reported to be associated with the efficiency of inhaled glucocorticoids in patients with asthma. This study aimed to investigate the role of GLCCI1 in the regulation of nucleotide-binding oligomerization domain (NOD)-like receptor (NLR) family pyrin domain-containing 3 (NLRP3) by the phosphatidylinositol 3-kinase (PI3K) pathway in the pathogenesis of allergic asthma.
The expression levels of genes encoding GLCCI1, NLRP3 inflammasome components, and PI3K pathway-related indicators were detected in cells isolated from induced sputum from patients with asthma and healthy controls. Next, we induced asthma in wild-type C57BL/6 mice and knockout ( ) mice by injecting them with ovalbumin (OVA) and treated the asthmatic mice with a PI3K pathway inhibitor (LY294002) or left them untreated. We also performed adoptive transfer of macrophages into the mice and assessed lung inflammation, as well as GLCCI1, PI3K pathway component, and NLRP3 inflammasome component expression levels. Finally, primary bone marrow-derived macrophages (BMDMs) from wild-type and mice were treated with OVA, either in the presence or absence of LY294002 and the NLRP3 inhibitor (MCC950), to validate our findings.
The mRNA level of in induced sputum cells from asthmatic patients was lower compared to that of healthy controls. Additionally, mRNA expression correlated negatively with NLRP3 inflammasome indicators and the PI3K pathway components, as well as with IL-1β expression in induced sputum macrophages. asthmatic mice showed elevated levels of airway inflammation and NLRP3 inflammasome activation compared to wild-type asthmatic mice. Surprisingly, the efficacy of LY294002 in reducing lung tissue inflammation and NLRP3 inflammasome activity in wild-type asthmatic mice was attenuated by knockout. LY294002 enhanced GLCCI1 levels in macrophages within the lung tissue of wild-type asthmatic mice. Moreover, LY294002 did not inhibit lung inflammation in wild-type asthmatic mice depleted of macrophages that had received adoptive transfer of BMDMs. experiments further illustrated that LY294002 suppressed NLRP3 activation by upregulating GLCCI1 expression in BMDMs. The introduction of MCC950 led to a marked decrease in NLRP3 and apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) protein levels, but did not affect the expression levels of GLCCI1 or the phospho-protein kinase B (p-AKT)/AKT ratio.
GLCCI1 deficiency promotes asthma inflammation through PI3K-induced NLRP3 inflammasome activation.
糖皮质激素诱导转录物1（GLCCI1）已被报道与哮喘患者吸入糖皮质激素的疗效相关。本研究旨在探讨GLCCI1在过敏性哮喘发病机制中通过磷脂酰肌醇3激酶（PI3K）途径对核苷酸结合寡聚化结构域（NOD）样受体（NLR）家族含pyrin结构域3（NLRP3）的调节作用。
检测哮喘患者和健康对照者诱导痰中分离的细胞中编码GLCCI1、NLRP3炎性小体成分和PI3K途径相关指标的基因表达水平。接下来，我们通过给野生型C57BL/6小鼠和敲除（ ）小鼠注射卵清蛋白（OVA）诱导哮喘，并使用PI3K途径抑制剂（LY294002）治疗哮喘小鼠或不进行治疗。我们还将巨噬细胞过继转移到小鼠体内，并评估肺部炎症以及GLCCI1、PI3K途径成分和NLRP3炎性小体成分的表达水平。最后，用OVA处理野生型和 小鼠的原代骨髓来源巨噬细胞（BMDM），同时存在或不存在LY294002和NLRP3抑制剂（MCC950），以验证我们的发现。
与健康对照相比，哮喘患者诱导痰细胞中 的mRNA水平较低。此外， mRNA表达与NLRP3炎性小体指标、PI3K途径成分以及诱导痰巨噬细胞中的IL-1β表达呈负相关。 哮喘小鼠与野生型哮喘小鼠相比，气道炎症水平和NLRP3炎性小体激活水平升高。令人惊讶的是，敲除 会减弱LY294002在减轻野生型哮喘小鼠肺组织炎症和NLRP3炎性小体活性方面的疗效。LY294002可提高野生型哮喘小鼠肺组织巨噬细胞中的GLCCI1水平。此外，LY294002对接受过继转移的 BMDM的野生型哮喘小鼠的肺炎症没有抑制作用。 实验进一步表明，LY294002通过上调BMDM中的GLCCI1表达来抑制NLRP3激活。引入MCC950导致NLRP3和含半胱天冬酶募集结构域的凋亡相关斑点样蛋白（ASC）蛋白水平显著降低，但不影响GLCCI1的表达水平或磷酸化蛋白激酶B（p-AKT）/AKT比值。
GLCCI1缺乏通过PI3K诱导的NLRP3炎性小体激活促进哮喘炎症。
