木犀草素对视网膜缺血-再灌注损伤(RIRI)大鼠TLR4/Syk/NF-κB信号通路及视网膜的影响  

Effects of luteolin on TLR4/Syk/NF-κB signaling pathway and retina in rats with retinal ischemia-reperfusion injury

作　　者：李茜[1] 王红星[1] 傅强 任泽钦[2] LI Qian;WANG Hongxing;FU Qiang;REN Zeqin(Department of Ophthalmology,Chuiyangliu Hospital Affiliated to Tsinghua University,Beijing 100022,China;Department of Ophthalmology,People’s Hospital of Peking University,Beijing 100000,China)

机构地区：[1]清华大学附属垂杨柳医院眼科,北京市100022 [2]北京大学人民医院眼科,北京市100000

出　　处：《眼科新进展》2021年第4期311-316,共6页Recent Advances in Ophthalmology

基　　金：北京市科学技术委员会科研计划项目(编号D181100000517009)。

摘　　要：目的探讨木犀草素对视网膜缺血-再灌注损伤(RIRI)大鼠Toll样受体4(TLR4)/脾脏酪氨酸激酶(Syk)/核因子-κB(NF-κB)信号通路及视网膜的影响。方法40只SD大鼠随机分为对照组、模型组、木犀草素低剂量组(5 mg·kg^(-1))、木犀草素高剂量组(10 mg·kg^(-1)),每组10只,除对照组外,其余各组大鼠制备RIRI模型,造模成功后分别进行腹腔注射不同试剂处理,每天1次,持续7 d。各组大鼠给药结束后24 h行光学相干断层扫描(OCT)检查,测量视盘周围视网膜厚度;取各组大鼠视网膜行HE染色观察视网膜组织病理形态、行TUNEL染色观察视网膜组织细胞凋亡情况;行酶联免疫吸附试验(ELISA)测定各组大鼠视网膜中超氧化物歧化酶(SOD)活性及丙二醛(MDA)水平,并检测大鼠视网膜中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平;利用Western blot检测各组大鼠视网膜TLR4/Syk/NF-κB信号通路相关蛋白表达水平。结果与对照组相比,模型组大鼠视网膜出现水肿、空泡变性及凋亡等损伤,视网膜厚度(156.31±18.76)μm、视网膜神经节细胞数(66.25±15.18)个·mm-1、SOD活性(50.33±1.98)U·mg^(-1)均显著降低(均为P<0.05),视网膜细胞凋亡率(21.36±2.04)%、视网膜中MDA(1.82±0.14)μmol·g^(-1)、TNF-α(98.42±11.25)g·L^(-1)、IL-6(87.34±7.62)g·L^(-1)含量及TLR4、p-Syk/Syk、p-NF-κB/NF-κB蛋白相对表达水平均显著升高(均为P<0.05)。与模型组相比,木犀草素低剂量组、木犀草素高剂量组大鼠视网膜水肿等损伤程度逐渐减轻,视网膜厚度(170.62±12.46)μm、(183.54±13.75)μm、视网膜神经节细胞数(86.74±16.21)个·mm-1、(105.44±14.23)个·mm-1、SOD活性(65.26±2.64)U·mg^(-1)、(73.48±3.13)U·mg^(-1)均依次增加(均为P<0.05),视网膜细胞凋亡率(16.75±1.55)%、(9.65±1.23)%、视网膜中MDA(1.23±0.12)μmol·g^(-1)、(0.86±0.09)μmol·g^(-1)、TNF-α(61.48±8.37)g·L^(-1)、(36.83±8.49)g·L^(-1)、IL-6(55.47±6.12)g·L^(-1)、(38.71±3.Objective To investigate the effects of luteolin on Toll-like receptor 4(TLR4)/spleen tyrosine kinase(Syk)/nuclear factor-κB(NF-κB)signaling pathway and retinal injury in rats with retinal ischemia-reperfusion injury.Methods SD rats(40)were randomly divided into control group,model group,luteolin low dose(5 mg·kg^(-1))group and luteolin high dose(10 mg·kg^(-1))group,with 10 rats in each group.Retinal ischemia-reperfusion rat models were established in model group,luteolin low dose group and luteolin high dose group.After successful modeling,the injection of different agents was carried out once a day for 7 days.Twenty-four hours after administration,optical coherence tomography(OCT)was used to measure the retinal thickness around the optic disc in rats;hematoxylin eosin staining(HE)was used to detect the pathological morphology of rat retina;apoptosis of retinal tissue cells was detected by TUNEL staining;the activity of superoxide dismutase(SOD)and the level of malondialdehyde(MDA)in retina were measured by enzyme-linked immunosorbent assay(ELISA);the levels of inflammatory factors tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were detected by ELISA;Western blot was used to detect the expression levels of TLR4/Syk/NF-κB signaling pathway related proteins in rat retina of groups.Results Compared with the control group,the retinal edema,vacuolar degeneration and apoptosis appeared in the model group,and the retinal thickness(156.31±18.76)μm,the number of retinal ganglion cells(66.25±15.18)cells·mm-1 and SOD activity(50.33±1.98)U·mg^(-1) were significantly decreased(all P<0.05),while the apoptosis index of retinal cells(21.36±2.04)%,MDA(1.82±0.14)μmol·g^(-1),TNF-α(98.42±11.25)g·L^(-1) and IL-6 levels(87.34±7.62)g·L^(-1),protein expression levels of TLR4,p-Syk/Syk,p-NF-κB/NF-κB were significantly increased(all P<0.05).Compared with the model group,the degree of retinal edema in luteolin low dose group and luteolin high dose group was gradually reduced,and the retinal thickness(170.62±

关 键 词：木犀草素 视网膜缺血-再灌注损伤 Toll样受体4/脾脏酪氨酸激酶/核因子-κB信号通路 视网膜损伤 氧化应激 

分 类 号：R774[医药卫生—眼科]