Upregulation of NLRP3 Inflammasome in the Tears and Ocular Surface of Dry Eye Patients.
Upregulation of NLRP3 Inflammasome in the Tears and Ocular Surface of Dry Eye Patients.
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干眼症患者泪液和眼表中 NLRP3 炎症小体的上调。
DOI：
10.1371/journal.pone.0126277
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Wang Y
中科院分区：
文献类型：
--
作者：
Niu L;Zhang S;Wu J;Chen L;Wang Y
目的探讨NLRP 3炎性小体及其下游炎症因子在干眼症眼组织中的mRNA和蛋白表达。我们招募了54例干燥综合征干眼（SSDE）患者，50例非干燥综合征干眼（NSSDE）患者和46例健康对照。对所有受试者进行泪膜破裂时间（TBUT）、Schirmer I试验和荧光素染色（FL）。通过酶联免疫吸附法（ELISA）分析泪液中IL-1β和IL-18的炎性细胞因子水平。收集结膜印迹细胞学（CIC）标本，采用定量RT-PCR检测NLRP 3、caspase-1、IL-1β、IL-18 mRNA表达，Western blotting检测NLRP 3、caspase-1蛋白表达。与对照组相比，两个干眼症组中NLRP 3 mRNA表达水平均较高，其中SSDE组显著升高（相对上调2.47倍，p<0.05）。与对照组相比，SSDE组NLRP 3蛋白表达也增加（相对上调1.94倍）。半胱天冬酶-1的mRNA表达在SSDE（相对1.44倍上调，p<0.05）和NSSDE（相对1.32倍上调，p<0.05）中均显著上调。与对照组相比，SSDE（相对1.84倍上调）和NSSDE（相对1.12倍上调）中的Procaspase-1蛋白水平增加;与对照组相比，SSDE（相对1.49倍上调）和NSSDE（相对1.17倍上调）中的caspase-1蛋白表达也增加。与对照组相比，SSDE和NSSDE患者IL-1β和IL-18 mRNA和蛋白表达水平均升高。与对照组相比，SSDE组和NSSDE组IL-1β mRNA相对表达量分别上调3.59倍（p<0.001）和2.13倍（p<0.01）。IL-1β蛋白水平在SSDE中也显示出显著上调（p=0.01;与对照组相比）。与对照组相比，SSDE（相对2.97倍上调，p=0.001）和NSSDE（相对2.05倍上调，p=0.001）组中IL-18 mRNA表达水平显著上调; SSDE（p<0.001）和NSSDE（p<0.05）组中泪液IL-18浓度也显著增加。在本研究中，我们发现NLRP 3炎性体的mRNA和蛋白表达在人类干眼中上调，尤其是在SSDE中;下游炎性因子caspase-1、IL-1β和IL-18在干眼患者中也升高。这些观察结果表明，NLRP 3炎性体参与干眼症炎症的发生和发展。
To evaluate the mRNA and protein expressions of NLRP3 inflammasome and its downstream inflammatory factors in human dry eye. We recruited 54 patients with Sjögren’s syndrome dry eye (SSDE), 50 patients with non-Sjögren’s syndrome dry eye (NSSDE), and 46 healthy controls. Tear film breakup time (TBUT), Schirmer I test, and fluorescein staining (FL) were performed on all subjects. Tear samples were obtained to analyze the inflammatory cytokine levels of IL-1β and IL-18 via enzyme-linked immunosorbent (ELISA). Conjunctival impression cytology (CIC) specimens were collected to detect the mRNA expression of NLRP3, caspase-1, IL-1β, and IL-18 using quantitative RT-PCR, and the protein expression of NLRP3 and caspase-1 by Western blotting. NLRP3 mRNA expression showed higher levels in both dry eye groups compared with controls, with a comparably significant elevation in the SSDE group (relative 2.47-fold upregulation, p<0.05). NLRP3 protein expression was also increased in SSDE group (relative1.94-fold upregulation) compared with the controls. mRNA expression of caspase-1 was significantly upregulated in both SSDE (relative 1.44-fold upregulation, p<0.05) and NSSDE (relative 1.32-fold upregulation, p<0.05). Procaspase-1 protein level was increased in SSDE (relative 1.84-fold upregulation) and NSSDE (relative 1.12-fold upregulation) versus controls; and caspase-1 protein expression was also increased in SSDE (relative 1.49-fold upregulation) and NSSDE (relative 1.17-fold upregulation) compared with the controls. The patients with SSDE and NSSDE had higher IL-1β and IL-18 mRNA values and protein expressions than the controls did. The relative mRNA expression of IL-1β upregulated 3.59-fold (p<0.001) in SSDE and 2.13-fold (p<0.01) in NSSDE compared with the controls. IL-1β protein level also showed significant upregulation in SSDE (p=0.01; vs. controls groups). IL-18 mRNA expression levels were significantly upregulated in the SSDE (relative 2.97-fold upregulation, p=0.001) and NSSDE (relative 2.05-fold upregulation, p=0.001) groups compared with the controls; tear IL-18 concentrations were also significantly increased in the SSDE (p<0.001) and NSSDE (p<0.05) groups. In the current study, we found that mRNA and protein expressions of NLRP3 inflammasome were upregulated in human dry eyes, especially in SSDE; the downstream inflammatory factors caspase-1, IL-1β, and IL-18 were also elevated in dry eye patients. These observations suggest the involvement of NLRP3 inflammasome in the onset and development of the inflammation in dry eye.
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影响因子：
3.4
作者：
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通讯作者：
Asbell, Penny A.
影响因子：
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Kaplan MJ
影响因子：
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作者：
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