[1]王俞方,肖启国,王智,等.ROS信号通路调控NLRP3炎症小体在自身免疫性葡萄膜炎发生中的作用[J].眼科新进展,2020,40(6):527-532.[doi:10.13389/j.cnki.rao.2020.0121]
　WANG Yufang,XIAO Qiguo,WANG Zhi,et al.Mechanism of ROS signaling pathway regulating NLRP3 inflammatory body in autoimmune uveitis[J].Recent Advances in Ophthalmology,2020,40(6):527-532.[doi:10.13389/j.cnki.rao.2020.0121]
点击复制

ROS信号通路调控NLRP3炎症小体在自身免疫性葡萄膜炎发生中的作用/HTML
分享到：

《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
40卷
期数:
2020年6期
页码:
527-532
栏目:
实验研究
出版日期:
2020-06-05

文章信息/Info

Title:
Mechanism of ROS signaling pathway regulating NLRP3 inflammatory body in autoimmune uveitis
作者:
王俞方; 肖启国; 王智; 周俊; 郑芳
421001　湖南省衡阳市，南华大学附属第二医院眼科
Author(s):
WANG Yufang; XIAO Qiguo; WANG Zhi; ZHOU Jun; ZHENG Fang
Department of Ophthalmology,the Second Hospital of University of South China,Hengyang 421001，Hunan Province，China
关键词:
自身免疫性葡萄膜炎; 活性氧簇; 信号通路; 核苷酸结合寡聚化结构域样受体
Keywords:
autoimmune uveitis;  reactive oxygen species;  signaling pathway;  nucleotide-binding and oligomerization domain-like receptor
分类号:
R773
DOI:
10.13389/j.cnki.rao.2020.0121
文献标志码:
A
摘要:
目的　探讨活性氧簇（ROS）信号通路调控核苷酸结合寡聚化结构域样受体（nucleotide binding and oligomerization domain-like receptor，NLRP3）炎症小体的表达在自身免疫性葡萄膜炎（autoimmune uveitis ，AU）发生中的作用。方法　健康大鼠共40只，随机分为空白对照组、模型组、空载体转染组（空载体组）和NLRP3-shRNA沉默组（沉默组）各10只；沉默组构建慢病毒载体pLenti6.3-EmGFP-NLRP3-shRNA，空载体转染组注射等量空载体，感染时间72 h。结果　ELISA结果发现，模型组和空载体组造模成功后10 d、13 d、16 d、19 d的NLRP3、IL-1β和IL-18水平显著高于沉默组，沉默组高于空白对照组（P<0.05）。免疫组织化学结果发现，模型组和空载体组12 d和20 d的NLRP3、凋亡相关斑点样蛋白（ASC）、pro-Caspase-1、IL-1β和IL-18阳性表达率显著高于沉默组，沉默组高于空白对照组（P<0.05）。PCR结果发现，模型组和空载体组造模成功后12 d和20 d的NLRP3 mRNA水平显著高于沉默组，沉默组高于空白对照组（P<0.05）。Western blot结果发现，模型组和空载体组造模成功后12 d和20 d的NLRP3、ASC、pro-Caspase-1、IL-1β和IL-18，硫氧还蛋白和硫氧还蛋白互作蛋白相对表达水平显著高于沉默组，沉默组高于空白对照组（P<0.05）。结论　NLRP3炎症小体及下游炎症因子表达上调可能参与了AU的发生，可能介导ROS信号通路发挥作用。
Abstract:
Objective　 To explore the effect of nucleotide binding oligomerization domain-like receptor (NLRP3) inflammatory body regulated by reactive oxygen species (ROS) signaling pathway in pathogenesis of autoimmune uveitis (AU).Methods　A total of 40 healthy rats were divided randomly into blank control group, model group, blank vector transfected group (blank vector group) and NLRP3-shRNA silencing group (silencing group), 10 rats in each group. The silencing group constructed lentiviral vector pLenti6.3-EmGFP-NLRP3-shRNA, the blank vector group injected the same amount of blank vector for 72 h. Results　 ELISA result showed that levels of NLRP3, IL-1β and IL-18 in model group and blank vector group were significantly higher than those in silencing group on 10th, 13th, 16th and 19th days, and were the lowest in blank control group (P<0.05). Immunohistochemical results showed that positive expression rates of NLRP3, apoptosis-related spot-like protein (ASC), pro-Caspase-1, IL-1β and IL-18 in model group and blank vector group on 12th and 20th days were significantly higher than those in silencing group, and were the lowest in blank control group (all P<0.05). PCR results showed that levels of NLRP3 mRNA in model group and blank vector group on 12th and 20th days were significantly higher than those in silencing group, and the lowest in blank control group (all P<0.05). Western blot results showed that relative expression levels of NLRP3, ASC, pro-Caspase-1, IL-1β and IL-18, thioredoxin and thioredoxin interacting protein in model group and blank vector group were significantly higher than those in silencing group, and were the lowest in blank control group on the 12th and 20th days (all P<0.05). Conclusion　 Up-regulation of NLRP3 inflammatory body and downstream inflammatory factors may be involved in occurrence of AU and may mediate ROS signaling pathway.

参考文献/References:

［1］　刘雪霞.葡萄膜炎的病因及治疗进展［J］.中华眼科医学杂志，2014，4(2):39-42.
LIU X X.Etiology and treatment progress of uveitis ［J］.Chin Med J Ophthalmol,2014,4 (2):39-42.
［2］　O’ROURKE M，FEARON U，SWEENEY C M，BASDEO S A，FLETCHER J M，MURPHY C C，et al.The pathogenic role of dendritic cells in non-infectious anterior uveitis［J］.Exp Eye Res，2018，173(8):121-128.
［3］　TEN BERGE J C，SCHREURS M W.Autoantibody profiling in intraocular fluid of patients with uveitis［J］.Exp Eye Res，2018，176(10):141-146.
［4］　HE Y，HARA H，NUNEZ G.Mechanism and regulation of NLRP3 inflammasome activation［J］.Trends Biochem Sci，2016，41(12):1012-1021.
［5］　POHLMANN D，SCHLICKEISER S，METZNER S，LENGLINGER M，WINTERHALTER S，PLEYER U.Different composition of intraocular immune mediators in Posner-Schlossman-Syndrome and Fuchs’Uveitis［J］.PLoS ONE，2018，13(6):e0199301.
［6］　JO E K，KIM J K,SHIN D M，SASAKAWA C.Molecular mechanisms regulating NLRP3 inflammasome activation［J］.Cell Mol Immunol，2016，13(2):148-159.
［7］　CHRISTIANNA C，GARYFALLIA P，ARGYRO R，KAMPOURAKI E，KAMBAS K，RITIS K，et al.Enhanced activity of NLRP3 inflammasome in peripheral blood cells of patients with active rheumatoid arthritis［J］.Arthrit Res Ther，2015，17(1):257.
［8］　JIJUN Z.Lupus nephritis:glycogen synthase kinase 3β promotion of renal damage through activation of the NLRP3 inflammasome in lupus-prone mice［J］.Arthrit Rheumatol，2015，67(4):1036-1044.
［9］　JIELI D，WANLIN L，TIANBO J，ZHAO Z，BAI R，XUE H，et al.A single-nucleotide polymorphism in MMP9 is associated with decreased risk of steroid-induced osteonecrosis of the femoral head［J］.Oncotarget，2016，7(42):68434-68441.
［10］　JIANG Y N，WANG H，YU H S，LI L，XU D，HOU S，et al.Two genetic variations in the IRF8 region are associated with Behcet’s disease in Han Chinese［J］.Sci Rep，2016，6(3):19651.
［11］　MINUTOLI L，PUZZOLO D，RINALDI M，IRRERA N，MARINI H，ARCORACI V，et al.ROS-mediated NLRP3 inflammasome activation in brain,heart,kidney,and testis ischemia/reperfusion injury［J］.Oxid Med Cell Longev，2016，6(4):1254-1255.
［12］　CHEN W，ZHAO M，ZHAO S，LU Q，NI L，ZOU C，et al.Activation of the TXNIP/NLRP3inflammasome pathway contributes to inflammation in diabetic retinopathy:a novel inhibitory effect of minocycline［J］.Inflamm Res，2017，66(2):157-166.
［13］　WANG Y F，PARPURA V.Astroglial modulation of hydromineral balance and cerebral edema［J］.Mol Neurosci，2018，204(6):2050-2062.
［14］　HAMANO M，HARAGUCHI Y，SAYANO T，ZYAO C，ARIMOTO Y，KAWANO Y，et al.Enhanced vulnerability to oxidative stress and induction of inflammatory gene expression in 3-phosphoglycerate dehydrogenase-deficient fibroblasts［J］.FEBS Open Bio，2018，8(5):914-922.
［15］　KLAUDIA K，GUNILLA T，WESTERMAR K，RENSTROM E，BLOMA M，KING B C.The human serum protein C4b-binding protein inhibits pancreatic IAPP-induced inflammasome activation［J］.Diabetologia，2017，60(8):1522-1533.
［16］　BURILLO-SANZ S，MONTES-CANO M A，GARCIA-LOZANO J R，ORTIZ-FERNANDEZ L，ORTEGO-CENTENO N，GARCIA-HERNANDEZ F J，et al.Mutational profile of rare variants in inflammasome-related genes in Behcet disease:A next generation sequencing approach［J］.Sci Rep，2017，7(1):8453.
［17］　MAJOR T J，DALBETH N，STAHL E A，MERRIMAN T R.An update on the genetics of hyperuricaemia and gout［J］.Nat Rev Rheumatol，2018，14(6):341-353.
［18］　KUMAR B，CASHMAN S M，KUMAR-SINGH R.Complement-mediated activation of the NLRP3 inflammasome and its inhibition by AAV-mediated delivery of CD59 in a model of uveitis［J］.Mol Ther，2018，26(6):1568-1580.
［19］　KURYLTSIV N B，HALEI K M.The role of interleukins and their inhibitors in the development of autoimmune uveitis［J］.Wiad Lek，2019，72(4):716-722.
［20］　CHEN J，CASPI R R.Clinical and functional evaluation of ocular inflammatory disease using the model of experimental autoimmune uveitis［J］.Methods Mol Biol，2019，1899(12):211-227.

相似文献/References:

[1]秦力维 高原 郭建巍.间充质干细胞治疗眼免疫性疾病的新进展[J].眼科新进展,2013,33(9):000.
[2]屈如意,周梦贤,毕宏生,等.巨噬细胞极化在眼科疾病发生发展过程中的作用机制研究进展[J].眼科新进展,2022,42(3):239.[doi:10.13389/j.cnki.rao.2022.0049]
　QU Ruyi,ZHOU Mengxian,BI Hongsheng,et al.Research advances in the role of macrophage polarization in the occurrence and development of ophthalmic diseases[J].Recent Advances in Ophthalmology,2022,42(6):239.[doi:10.13389/j.cnki.rao.2022.0049]

备注/Memo

备注/Memo:
湖南省衡阳市科技局资助项目（编号：S2018F9031022227）
更新日期/Last Update: 2020-06-05