PDCD4沉默通过调控NLRP3/NLRP6炎症小体的平衡来减轻视网膜缺血再灌注损伤后的神经炎症反应  

PDCD4 deficiency reduces neuroinflammatory response after retinal ischemia-reperfusion injury by regulating the balance of NLRP3/NLRP6 inflammasome

作　　者：魏丽 刘晓环 王连婷 朱慧颖 王晓蓉 WEI Li;LIU Xiaohuan;WANG Lianting;ZHU Huiying;WANG Xiaorong(Department of Ophthalmology,Mudanjiang Hospital of Traditional Chinese Medicine,Mudanjiang,Heilongjiang 157000,China;Heilongjiang Eye Disease Prevention and Treatment Institute,Harbin,Heilongjiang 150001,China)

机构地区：[1]黑龙江省牡丹江市中医医院眼科,黑龙江牡丹江157000 [2]黑龙江省眼病防治所,黑龙江哈尔滨150001

出　　处：《检验医学与临床》2024年第10期1461-1467,共7页Laboratory Medicine and Clinic

摘　　要：目的探究程序性细胞死亡因子4(PDCD4)通过含NOD样受体家族Pyrin域蛋白(NLRP)3/NLRP6信号通路活性对视网膜缺血再灌注(RIR)损伤后神经炎症反应的抑制作用及机制。方法选取50只SD大鼠作为研究对象,根据不同处理方式,分为Sham组(空白对照组)、RIR模型组、MCC950组(NLRP3/NLRP6抑制组)、si-PDCD4组(PDCD4沉默组)、si-PDCD4+MCC950组(PDCD4沉默+NLRP3/NLRP6抑制组),每组10只大鼠。除去空白对照组大鼠外,剩余大鼠均构建RIR损伤模型,并进行相应处理;Western-blot检测视网膜组织内PDCD4、NLRP3、NLRP6、天冬氨酸蛋白水解酶-1(Caspase-1)、抗坏血酸过氧化物酶(ASC)表达水平;苏木精-伊红(HE)染色分析视网膜组织病理变化;终端尿苷酸核苷酸末端标记法检测视网膜组织细胞凋亡能力;酶联免疫吸附试验检测大鼠眼球血、视网膜组织内炎症因子白细胞介素(IL)-6、IL-18、IL-1β、肿瘤坏死因子(TNF)-α水平;Western-blot检测视网膜组织内细胞凋亡相关蛋白Bax、Bcl-2、Cleaved-caspase-3表达水平。结果与Sham组比较,RIR模型组PDCD4、NLRP3、NLRP6、Caspase-1、ASC、IL-6、IL-8、IL-1β、TNF-α水平及细胞凋亡指数(AI)均升高,Bcl-2表达水平降低,差异有统计学意义(P<0.05);与RIR模型组比较,MCC950组、si-PDCD4组大鼠NLRP3、NLRP6、Caspase-1、ASC、IL-6、IL-8、IL-1β、TNF-α水平及AI均降低,Bcl-2表达水平升高,差异有统计学意义(P<0.05);与si-PDCD4组及MCC950组比较,si-PDCD4+MCC950组PDCD4、NLRP3、NLRP6、Caspase-1、ASC、IL-6、IL-8、IL-1β、TNF-α水平及AI进一步降低,Bcl-2表达水平进一步升高,差异有统计学意义(P<0.05)。结论PDCD4沉默具有减轻RIR病理损伤,抑制视网膜细胞凋亡的作用,其分子机制可能与抑制NLRP3/NLRP6炎症小体诱导神经炎症反应有关。Objective Exploring the role of programmed cell death factor 4(PDCD4)in regulating NOD like receptor thermal protein domain associated protein(NLRP)3/NLRP6 for the inhibitory effect and mechanism of signal pathway activity on neuroinflammatory response after retinal ischemia-reperfusion(RIR)injury.Methods Fifty SD rats were divided into Sham group(blank control group),RIR model group,MCC950 group(NLRP3/NLRP6 inhibition group),si-PDCD4 group(PDCD4 silencing group),si-PDCD4+MCC950 group(PDCD4 silencing+NLRP3/NLRP6 inhibition group),with 10 rats in each group.Except for rats in the blank control group,the remaining rats were established retinal ischemia-reperfusion(RIR)injury model and treated accordingly.The protein expressions of PDCD4,NLRP3,NLRP6,Caspase-1 and ascorbate peroxidase(ASC)in retinal tissue were detected by Western-blot.Hematoxylin-eosin(HE)staining was used to analyze the pathological changes of retinal tissue.Terminal transferase uridyl nick end labeling(TUNEL)method was used to detect the apoptosis of retinal cells.Enzyme-linked imunosorbent assay was used to detect the levels of inflammatory factors interleukin(IL)-6,IL-18,Il-1βand tumor necrosis factor(TNF)-αin eyeball blood and retinal tissue.The expressions of apoptosis-related proteins Bax,Bcl-2 and Cleaved-caspase-3 in retinal tissue were detected by Western-blot.Results Compared with the Sham group,the levels of PDCD4,NLRP3,NLRP6,Caspase-1,ASC,IL-6,IL-8,IL-1βand TNF-αand the index of apoptosis(AI)increased,while the expression of Bcl-2 decreased in the RIR model group(P<0.05).Compared with those in the RIR model group,the levels of NLRP3,NLRP6,Caspase-1,ASC,IL-6,IL-8,IL-1β,TNF-αand AI decreased,while the expression of Bcl-2 increased in MCC950 group and si-PDCD4 group,the differences were statistically significant(P<0.05).Compared those in the MCC950 group and si-PDCD4 group,the levels of PDCD4,NLRP3,NLRP6,Caspase-1,ASC,IL-6,IL-8,IL-1βand TNF-αand the AI were further down-regulated,and the expression of Bcl-2 was further up-regulated

关 键 词：程序性细胞死亡因子4 NLRP3/NLRP6炎症小体 视网膜缺血再灌注损伤 神经炎症 白细胞介素 肿瘤坏死因子-α 

分 类 号：R774[医药卫生—眼科]