内毒素诱导的葡萄膜炎大鼠眼内DR5的表达研究

目的 观察内毒素诱导的葡萄膜炎动物模型眼内DR5的表达,研究炎症细胞的凋亡与TRAIL/DR5表达的关系.方法 选取SD大鼠随机分为3组:空白对照组、生理盐水注射组、内毒素注射组.其中内毒素注射组是将内毒素注射到大鼠后足底制成葡萄膜炎动物模型.空白对照组无任何操作,生理盐水注射组以及内毒素注射组又根据注射后不同时间分为6h、12 h、24 h、48 h4个亚组.在注射后不同时间摘除眼球,通过透射电镜观察虹膜毛细血管炎症细胞和内皮细胞的超微结构变化;运用免疫组织化学法检测内毒素诱导后不同时间DR5蛋白在炎症细胞上的表达.结果 透射电镜显示:空白对照组以及生理盐水注射组虹膜组织中毛细血管内皮细胞可见微绒毛、较多的吞饮小泡,未见有明显的结构、形态异常.内毒素注射6h组的毛细血管内皮细胞的吞饮小泡数目开始减少,浸润的中性粒细胞和淋巴细胞出现了染色质边集的表现;24 h组吞饮小泡数目减少明显,大量的中性粒细胞和淋巴细胞出现了染色质浓缩、线粒体空泡样变的凋亡表现.免疫组织化学结果显示:DR5蛋白在空白对照组以及生理盐水注射组大鼠的虹膜色素上皮层呈阴性着色;内毒素注射组中,DR5蛋白在虹膜组织的炎症细胞上呈阳性着色,从6h组炎症细胞就出现,24 h组DR5在炎症细胞中的着色数量及着色强度达到最高,光密度值达到0.085 9±0.019 6,与6h组、12 h组、48 h组比较差异均有统计学意义(均为P＜0.05).结论 TRAIL及其受体DR5可能参与了内毒素诱导的葡萄膜炎中炎症细胞的凋亡.     

内毒素诱导的大鼠葡萄膜炎细胞凋亡的研究

目的探讨大鼠内毒素诱导的葡萄膜炎（EIU）中葡萄膜和视网膜组织浸润细胞的凋亡,及Fas-FasL的表达与细胞凋亡的相关性。方法注射霍乱弧菌内毒素诱发大鼠EIU模型,内毒素注射后6、12、18、24、48h进行临床观察评分并行组织病理学检查。应用TUNEL法检测葡萄膜和视网膜中的凋亡细胞,免疫组织化学法检测Fas和FasL在葡萄膜和视网膜中的表达和变化。结果实验组大鼠均发生了EIU;葡萄膜和视网膜组织浸润细胞中可见TUNEL染色阳性凋亡细胞,18h达高峰,以虹膜和睫状体最为明显,凋亡阳性细胞数与正常对照组相比,差异有统计学意义（P〈0.01）;实验各组中均可见Fas表达,实验各组中FasL表达显著增强,与正常对照组相比差异有统计学意义（P〈0.01）。Fas和FasL的表达与凋亡之间呈正相关（r=0.923,0.807,P〈0.01）。结论EIU中葡萄膜和视网膜的浸润细胞存在凋亡现象,凋亡参与了炎症的迅速消退,Fas和FasL的高表达可能与内毒素诱导的葡萄膜炎眼组织的细胞凋亡有关。     

内毒素耐受对内毒素诱导的葡萄膜炎炎症程度及虹膜睫状体中PI3K/AKT 信号通路的影响

目的探讨低剂量脂多糖(lipopolysaccharide,LPS)预处理对内毒素诱导葡萄膜炎(endotoxin induced uveitis,EIU)动物模型炎症程度的影响,并分析虹膜睫状体组织中PI3K/AKT表达水平的改变。设计实验研究。研究对象8-10周龄健康雄性Wistar大鼠90只。方法随机分为正常对照(normal control,NC)组、内毒素耐受(endotoxin tolerance,ET)组和内毒素诱导的葡萄膜炎(EIU)组,每组30只。通过皮下注射1 mg/kg LPS建立内毒素诱导的葡萄膜炎动物模型。ET组:EIU模型诱导之前连续5日腹腔注射0.1 mg/kg LPS诱导内毒素耐受。EIU组及NC组连续5日注射等体积生理盐水。第6天相同时间ET组及EIU组接受200μg LPS皮下注射,NC组接受等体积生理盐水注射。注射后24小时,在裂隙灯显微镜下进行临床评分。眼球摘除行HE染色组织病理学观察。ELISA测量房水TNF-α浓度,蛋白印迹法检测虹膜睫状体PI3K和AKT蛋白的表达。主要指标眼前节炎症反应评分,虹膜切片HE染色观察,房水TNF-α浓度及虹膜睫状体PI3K/AKT蛋白表达。结果临床评分ET组、EIU组、NC组分别为(1.65±0.49)、(6.65±0.59)、(0.20±0.41)分(χ~2=53.261,P=0.001);组织病理评分ET组、EIU组、NC组分别为(0.6±0.5)、(3.8±0.4)、(0.2±0.4)分(χ~2=46.137,P=0.001)。房水TNF-α浓度ET组、EIU组、NC组分别为(10.58±0.67)、(30.96±1.55)、(10.32±0.61)pg/ml(F=260.08,P=0.03)。蛋白印迹法显示虹膜睫状体组织中PI3K和AKT的蛋白水平ET组、EIU组、NC组分别为(0.197±0.019)、(0.539±0.017)、(0.453±0.014)(F=210.66,P=0.002)和(0.234±0.019)、(0.553±0.013)、(0.428±0.002)(F=275.35,P=0.001)。结论内毒素耐受可以降低大鼠内毒素诱导的葡萄膜炎的炎症反应程度,下调虹膜睫状体组织中PI3K和AKT的表达,揭示内毒素耐受对EIU的保护机制可能与PI3K/AKT信号通路有关。     

红芪多糖干预内毒素诱导的大鼠葡萄膜炎的机制

目的　探讨红芪多糖（hedysarum polybotrys saccharide,HPS）在内毒素诱导的大鼠葡萄膜炎发病中的作用及对Toll样受体4（Toll-like receptor 4,TLR-4）信号转导通路中关键分子肿瘤坏死因子受体相关因子6（tumor necrosis factor receptor-associated factor 6，TRAF6）及TIR功能区接头蛋白诱导的干扰素β（TIR-domain-containing adapter-inducing interferon-β，TRIF)表达的影响。方法　选用SPF级健康成年Wistar大鼠40只，随机分为内毒素诱导的葡萄膜炎（endotoxin induced uveitis,EIU）模型组、HPS组、HPS+内毒素组、正常对照组（normal control，NC）组，每组10只。EIU模型组大鼠采用皮下注射1 mg·kg^-1内毒素的方法建立大鼠急性前葡萄膜炎动物模型。HPS+内毒素组在内毒素注射前1 h给予腹腔注射HPS 400 mg·kg^-1。HPS组大鼠只注射HPS 400 mg·kg^-1。每2 h用裂隙灯观察大鼠眼前节炎症反应，注射后的0 h、6 h、12 h、18 h、24 h对各组眼部临床炎症反应进行评分。注射后24 h组织病理学检查炎症反应水平。RT-PCR检测核因子-κB、TRAF6及TRIF mRNA的表达。结果　NC组大鼠24 h内眼前节未见炎症反应。注射后24 h,EIU模型组可见虹膜血管扩张明显，瞳孔区大量白色渗出,HPS组仅可见虹膜血管扩张，未见其他眼部炎症反应,HPS+内毒素组大鼠可见虹膜血管扩张和瞳孔缘少量渗出。比较不同组间的大鼠眼部临床炎症反应评分可见，NC组与EIU模型组、HPS+内毒素组、HPS组比较，差异均有统计学意义(均为P＜0.001）；EIU模型组与HPS+内毒素组、HPS组比较，差异均有统计学意义（均为P<0.05）;HPS+内毒素组与HPS组比较，差异有统计学意义（P＜0.001）。RT-PCR结果显示，HPS可降低核因子-κB mRNA和TRAF6 mRNA的表达（P＜0.001），HPS对TRIF的表达无明显影响（P=0.236）。结论　HPS可能通过抑制TRAF6核酸的表达缓解EIU的炎症反应,HPS对TRIF的表达无明显影响。     

人红细胞保护蛋白提纯及其对葡萄膜炎的作用

目的 探讨人红细胞保护蛋白(PRP)对内毒素诱导葡萄膜炎(EIU)的影响。方法 用等量饱和硫酸铵溶液从人新鲜红细胞中盐析出基质可溶性蛋白,进行CM-Sephadex C50阳离子交换和DEAE Sephacel阴离子交换层析,以提纯PRP。6只PRP处理SD大鼠和4只未处理SD大鼠均于足底部注射200μg鼠伤寒杆菌内毒素(LPS),以诱导EIU,每只PRP处理鼠在LPS注射同时及注射前2h腹腔注射0.25mg还原型PRP,于LPS注射后24h评价两组大鼠临床表现、房水蛋白浓度、房水细胞数及虹膜睫状体平片上ED1~+细胞数等。结果 从30ml人新鲜红细胞中可提纯15.3mg的PRP,其纯度为99％;PRP处理组大鼠在临床表现积分、房水蛋白质量浓度、房水细胞数和虹膜睫状体平片上ED1~+细胞数等方面都显著低于未处理组(P<0.05)。结论 用CM-Sephadex C50阳离子交换和DEAE Sephacel阴离子交换层析的方法能从红细胞中提取高产量、高纯度的PRP;PRP对EIU有预防和治疗作用。     

内毒素诱导的SD大鼠葡萄膜炎外周血淋巴细胞凋亡的研究

目的探讨外周血淋巴细胞凋亡在内毒素诱导的SD大鼠葡萄膜炎模型炎症过程中的作用.方法将内毒素注射于SD大鼠的足底部,于注射后不同时间点抽取实验组鼠的外周血,分离淋巴细胞并制备涂片,分别用流式细胞仪和TUNEL法检测凋亡淋巴细胞的阳性百分率.结果临床观察和组织学检查结果均证实50只SD大鼠全部发生了葡萄膜炎,以前葡萄膜炎为主,炎症平均得分为4.589级±1.095级;检测结果显示在炎症高峰期(24～36h)实验组鼠外周血淋巴细胞发生大量凋亡.结论内毒素诱导外周血淋巴细胞的凋亡可能参与了炎症的迅速消退.     

TLR4-MyD88在大鼠急性前葡萄膜炎虹膜中的表达

目的观察内毒素诱导的大鼠急性前葡萄膜炎（EIU）虹膜组织内Toll样受体4（TLR4）、髓样分化因子88（MyD88）及核因子-κBp65（NF-κB p65）的表达。方法Wistar大鼠50只,随机分为5组,0、12、24、48、72h,每组10只。0h组为正常对照组,其余4组均足垫部注射霍乱弧菌内毒素脂多糖（LPS）200μg,建立EIU动物模型,每隔2h用裂隙灯观察大鼠眼前节炎症反应。通过铺片免疫组织化学染色,检测虹膜睫状体组织内TLR4、MyD88和NF—κB p65的表达,并对虹膜内TLR4^＋和MyD88^＋及NF—κB p65^＋细胞进行计数。结果注射后24～48h大鼠眼前段的炎症反应达到高峰,72h炎症反应逐渐缓解。组织病理学检查表明,虹膜睫状体组织的炎性细胞浸润在24～48h达到高峰,与临床反应结果相符。TLR4在模型鼠虹膜睫状体炎复合体中表达的免疫组织化学检测结果表明,0h组虹膜铺片内无阳性细胞,12h后可见细胞形态大多为类圆形的阳性细胞,48h达高峰,72h阳性细胞数开始减少,各组阳性细胞数总体差异有统计学意义（F=46．79,P〈0．05）。MyD88和NF—κB p65的表达与TLR4的改变趋势相一致（F=54．37,P〈0．05;F=85．32,P〈0．05）。结论内毒素诱导的EIU虹膜内,TLR4及其下游信号传导分子的表达量发生改变,提示TLR4-MyD88依赖传导途径可能参与了EIU的发病.     

霍乱弧菌内毒素诱导大鼠葡萄膜炎的组织切片研究

目的：本研究用霍乱弧菌内毒素诱导大鼠葡萄膜炎，建立全葡萄膜炎的动物模型，以期研究该病的发病机制和有效的治疗方案，方法：应用纯化后霍乱弧菌内毒素注射于Wistar大鼠，分别于注射后4，8，12，16，24h裂隙灯显微镜观察以以眼前节变化，及病理组织学观察，结果；注射内毒素后4h出现炎症，12h反应达到最重、表现为瞳孔缩小，虹膜水肿，血管充血扩张，严重者前房渗出，积血，组织学观察可见前房睫状体，虹膜水肿血管扩张充血，脉络膜水肿加剧，血细胞弥漫性渗出，视网膜有脱离，节细胞数量，视网膜厚度，节细胞大小，节细胞层血管数量及大小，内核外网层血管数量及大小都有明显的变化，结论：认为该方法用于诱导葡萄膜炎动物模型是成功的，同时还比较了内毒素和S抗原在诱导葡萄膜炎中的关系。     

MMP-9及TIMP-1在内毒素诱导性葡萄膜炎大鼠中的表达

目的 探讨基质金属蛋白酶-9(MMP-9)及其组织型抑制剂(tissue inhitor of metallopoteinase-1，TIMP—1)在内毒素诱导性葡萄膜炎(endotoxin induced uveitis,EIU)模型中的表达及意义。方法 200μg伤寒杆菌内毒素注射于SD大鼠双后足垫。建立EIU模型。在内毒素注射后的不同时间点处死大鼠。采用免疫组织化学方法和计算机图像分析系统检测MMP-9和TIMP-1在不同时间点的表达及其积分吸光度(A)值。结果内毒素注射6h开始出现炎症，以后炎症加重。于24h达到炎症高峰，以后炎症减轻。7d时基本消退。虹膜上皮细胞，睫状体上皮细胞和渗出的炎症细胞表达MMP-9和TIMP-1。MMP-9在6h出现表达，18～24h达高峰，以后逐渐下降，7d消失。TIMP—1于24h出现表达，72h达高峰，以后逐渐下降。结论 大鼠EIU模型炎症早期MMP-9活性增高，继而TIMP-1分泌增多，MMP-9活性受抑，炎症减轻。提示MMP-9可能是参与EIU的重要调控因子，而TIMP-1则在炎症的抑制过程中发挥了重要作用。     

Toll样受体-4在内毒素诱导的葡萄膜炎中的表达及意义

目的 探讨Toll样受体-4(TLR4)在内毒素诱导的葡萄膜炎中的表达和意义.设计 实验性研究.研究对象 Wistar大鼠12只,随机分为模型组(n=6)和对照组(n=6).方法 模型组通过足底及腹腔注射霍乱弧菌内毒素诱导出葡萄膜炎,对照组注射磷酸盐缓冲液.注射后4h、10h、16h、24h裂隙灯观察眼前节反应.注射后24h通过免疫组化方法检测眼球冰冻切片及葡萄膜铺片中TLR4的表达.TLR4阳性判定标准:呈棕黄色颗粒定位于胞膜、胞浆.主要指标 眼前节炎症反应程度、葡萄膜铺片中TLR4阳性表达的细胞数量.结果 内毒素注射后4h虹膜血管扩张充血,16h前房可见纤维素渗出.模型组虹膜铺片内可见大量TLR4阳性表达细胞,位于虹膜基质层,对照组虹膜铺片内只见少量TLR4阳性表达细胞,模型组虹膜中阳性细胞数量高于对照组(P＜0.05).冰冻切片中虹膜内偶见少量TLR4表达.脉络膜和视网膜冰冻切片及铺片中均未见阳性细胞.结论 内毒素诱导的葡萄膜炎中TLR4表达增高,提示TLR4可能在急性前葡萄膜炎的发生发展中具有一定作用.(眼科,2008,17:48-51)     

内毒素诱导性葡萄膜炎眼中NO含量和基质金属蛋白酶-9及其组织型金属蛋白酶抑制剂-1与诱导型NO合酶的表达

目的 观察基质金属蛋白酶（MMP）-9及其组织型金属蛋白酶抑制剂（TIMP）-1、一氧化氮（NO）和诱导型一氧化氮合酶（iNOS）在内毒素诱导性葡萄膜炎（EIU）眼组织中的变化。 方法 90只Sprague-Dawley(SD)大鼠随机分为实验组(81只)和对照组(9只)。实验组大鼠双后足垫注射伤寒杆菌内毒素(LPS)200 μl建立EIU模型；对照组大鼠不予注射。在LPS注射后0、6、12、18、24、48、72、96h和7d分别处死9只实验组大鼠，观察眼部改变并进行组织病理学检查。检测血浆、房水和葡萄膜组织的NO含量和房水蛋白浓度。免疫组织化学方法和计算机图像分析系统检测MMP-9、TIMP-1和iNOS的表达及其平均吸光度［A, 旧称光密度（OD）］值。 结果 虹膜、睫状体的上皮细胞和渗出的炎性细胞表达iNOS、MMP-9和TIMP-1。房水蛋白浓度，血浆、房水和葡萄膜组织中NO含量，以及MMP-9的A值与炎症程度呈正相关，iNOS和TIMP-1的A值与炎症程度无明显相关性。iNOS在LPS注射后6h可见表达，12h达高峰，以后逐渐下降。TIMP-1表达出现于24h，72h时达高峰。 结论 在EIU发生、发展过程中, MMP-9、TIMP-1、NO、iNOS的含量或表达发生变化，提示它们参与EIU的病理过程。 (中华眼底病杂志, 2005, 21: 371-374)     

Visualization of cell death in vivo during murine endotoxin-induced uveitis

PURPOSE: To develop a technology to image cell death in the eye of a live mouse and to apply that technology to characterize the role of apoptosis and necrosis in the evolution of endotoxin-induced uveitis (EIU), a standard model of intraocular inflammation. METHODS: To induce EIU, 250 ng Escherichia coli 055:B5 lipopolysaccharide was injected into the vitreous body of BALB/c mice. At 0, 6, 10, 16, 20, 24, 48, 72, and 96 hours and on day 7 after endotoxin injection, annexin V and propidium iodide were injected into the anterior chamber of these mice, and labeled cells were observed by using intravital epifluorescence video microscopy. Iris and corneal wholemounts isolated from the mice were also evaluated with standard and confocal fluorescence microscopy. TUNEL staining was performed on iris wholemounts taken from additional mice similarly injected with endotoxin, to confirm the in vivo results. RESULTS: Uveitis was induced in all the mice that received an endotoxin injection. The percentages of annexin V(+) propidium(-) cells, annexin V(+) propidium(+) cells, and annexin V(-) propidium(+) cells in the iris tissues visible by intravital microscopy were comparable to those observed by TUNEL staining in vitro. In addition, intravital microscopy allowed observation of labeled cells in the aqueous humor and on the surface of the lens. Both the number and the pattern of labeled cells changed dramatically over time. The cells stained with annexin V had a variety of morphologies, including small and round, round with a lobulated or a kidney-shaped nucleus, dendriform, and irregular. CONCLUSIONS: A technique was developed to image cell death in the anterior segment of the eye in vivo and used to demonstrate that the number and proportion of early apoptotic (annexin V(+) propidium(-)) and late apoptotic or necrotic (annexin V(+) propidium(+) and annexin V(-) propidium(+)) cells change over the course of EIU. A variety of inflammatory cells and resident cells undergo apoptosis, or possibly necrosis, which may contribute to the rapid resolution of EIU. This in vivo technique will be a valuable tool for future studies on the resolution of ocular inflammation.     

Endotoxin-induced uveitis in mice. 1. Induction of uveitis and role of T lymphocytes.

Endotoxin-induced uveitis (EIU) with a high frequency of posterior iris synechiae was induced by the systemic injection of 200 micrograms of endotoxin into C3H/HeN mice, an endotoxin-responsive strain. The cell number and the protein concentration within the aqueous humor began to increase 6 hours after the injection, achieving a peak at 24 hours, and decreased gradually thereafter. Inflammatory cells were observed in the anterior chamber, the vitreous body and near the iris-ciliary body histologically. Most of the inflammatory cells were polymorphonuclear cells. On the other hand, C3H/HeJ mice, an endotoxin-unresponsive strain, showed no increase in either cell number or protein concentration in the aqueous humor after endotoxin administration. Pretreatment of C3H/HeN mice with anti-Thy-1.2 antibody significantly decreased both the cell number and the protein concentration in the aqueous humor and the incidence of the posterior synechiae, as compared with the control group. Anti-CD4 antibody also significantly reduced the severity of EIU, while anti-CD8 antibody had no influence on the disease. Anti-IFN-gamma antibody increased the cell number in the aqueous humor. These observations indicate that T lymphocytes, especially CD4+ T lymphocytes, have an extremely important role in the development of EIU in mice.     

Recurrent intraocular inflammation in endotoxin-induced uveitis

PURPOSE: Endotoxin-induced uveitis (EIU) in rats and mice peaks 24 hours after endotoxin injection and is commonly assumed to be a monophasic disease. This study examined intraocular inflammation at later time points to determine whether endotoxin injection can induce recurrent intraocular inflammation in strains of mice with high or moderate levels of susceptibility to EIU. METHODS: EIU was elicited in two mouse strains with high (C3H/HeN) and moderate (FVB/N) susceptibility, by means of intraperitoneal injections of Salmonella typhimurium endotoxin. Inflammatory cells in the anterior and posterior segments of the eye were counted by a masked observer on histologic sections of eyes from 1 to 17 days after endotoxin injection. RESULTS: A bimodal distribution of inflammatory cell infiltration was noted in eyes from C3H/HeN mice. As previously reported, inflammation peaked at 24 hours after endotoxin injection. However, a second, more pronounced peak of intraocular inflammation occurred approximately 5 days after endotoxin injection. FVB/N mice had a single peak of intraocular inflammation 4 days after injection. CONCLUSIONS: Endotoxin injection in C3H/HeN elicits recurrent intraocular inflammation. The previously unrecognized second peak of inflammation is more severe than the initial inflammatory disease. Studies on this second inflammatory peak may be useful in determining the pathogenesis of recurrent uveitis in humans.     

Endotoxin-induced uveitis is partially inhibited by anti-IL-8 antibody treatment.

PURPOSE: To examine the potential therapeutic effect of a neutralizing anti-IL-8 monoclonal antibody in endotoxin-induced uveitis (EIU) in the rabbit. METHODS: An anti-IL-8 antibody (WS-4) was injected intravitreal 2 hours before, simultaneously with, or 6 hours after endotoxin challenge in rabbits. Eyes were examined for clinical signs of inflammation, and aqueous humor (AH) was sampled to study cellular infiltration and protein content. Leukocyte subset analysis was performed on Giemsa-stained AH cytospins. Histologic grading of inflammation was performed on hematoxylin-eosin-stained sagittal sections of enucleated eyes. In separate experiments, animals received the anti-IL-8 antibody simultaneously with the endotoxin challenge, before repeated anterior chamber paracentesis was performed (at 6, 12, 24, 48, and 72 hours after injection) to estimate the kinetics and durability of changes in total cell count and protein concentration in AH. RESULTS: Anti-IL-8 therapy caused a decrease in the clinical and histologic grade of inflammation in EIU. The mean cell count in the AH at the peak of inflammation (24 hours) in eyes receiving endotoxin only was 6419+/-1165/microl (mean +/- SE) compared to 2546+/-573/microl in rabbits treated simultaneously with 250 microg of anti-IL-8 antibody (P < 0.05). The protein concentration in the AH was not significantly altered by anti-IL-8 treatment. Kinetic analysis of the leukocyte count in the AH demonstrated persistent inhibition of leukocyte accumulation (range, 60%-91% compared to control eyes) by the anti-IL-8 antibody administered simultaneously with endotoxin. This inhibition was sustained for up to 72 hours after injection. CONCLUSIONS: Anti-IL-8 antibody treatment partially blocks EIU in rabbits. A consistent decrease in the recruitment of polymorphonuclear leukocytes into the anterior chamber was obtained when neutralizing antibody was injected simultaneously with endotoxin. These findings suggest that IL-8 contributes to the chemotactic signal for the recruitment of leukocytes in EIU.     

High-dose infliximab prophylaxis in endotoxin-induced uveitis.

PURPOSE: The aim of this study to analyze the preventive effect of high-dose infliximab in endotoxin-induced uveitis (EIU) in rabbits. METHODS: An experimental study was conducted on 64 rabbits. Salmonella typhimurium lipopolysaccharide endotoxin was intravitreally injected. Infliximab was intravenously (i.v.) injected 24 h before the intravitreal injection (20 mg/kg). The animals were randomly assigned to five groups: group A, saline intravitreal injection; group B, Infliximab i.v. group C, infliximab + saline; group D, intravitreal endotoxin and group E, infliximab i.v. + intravitreal endotoxin. With two masked observers, a microscopic examination of aqueous humor (cells, tumor necrosis factor [TNF] alpha) and aqueous protein level were performed 24 h after an endotoxin injection and 48 h after an infliximab infusion. RESULTS: Infliximab treatment, at a dose of 20 mg/kg, significantly improved all the parameters. Inflammatory cell infiltration was significantly reduced in the iris, ciliary body, and anterior chamber (U Mann-Whitney test, P = 0.01). Associated with a lower level of TNF-alpha and protein exudate in aqueous humor (U Mann-Whitney test, P = 0.01). CONCLUSIONS: Infliximab, at a dose of 20 mg/kg, is effective in the prophylaxis of the EIU.     

Systemic interferon-gamma suppresses the development of endotoxin-induced uveitis in mice

PURPOSE: It has been shown that interferon (IFN)-gamma is involved in the development of endotoxin-induced uveitis (EIU), but its exact role is unclear. We aimed to elucidate the role that endogenous systemic IFN-gamma plays in EIU pathogenesis. METHODS: EIU was induced in wild-type (WT) or IFN-gamma knockout (GKO) mice on the C57BL/6 background by injecting Salmonella typhimurium endotoxin into a hind footpad. Twenty-four hours later, the eyes were harvested for histological analysis, and the serum was collected for cytokine ELISAs. WT and GKO mice were also intraperitoneally injected with 1 microg of recombinant murine IFN-gamma (rmIFN-gamma) just after and 6 h after EIU induction, and their eyes and sera were evaluated 24 h after EIU induction, as above. RESULTS: The GKO mice had significantly more severe EIU as determined by the number of ocular infiltrating cells and lower serum IL-6 levels after EIU induction compared to WT mice. The injection of rmIFN-gamma suppressed the severity of EIU and increased the serum IL-6 levels in both the WT and GKO mice. CONCLUSIONS: Endogenous IFN-gamma suppresses EIU pathogenesis. In addition, the systemic administration of IFN-gamma suppresses EIU. The suppressive mechanism involved is unclear but may relate to the production of IL-6.     

Anti-Inflammatory Effect of Dehydroxymethylepoxyquinomicin,a Nuclear factor–κB Inhibitor,on Endotoxin-Induced Uveitis in Rats In vivo and In vitro

ABSTRACT

Purpose: To determine the anti-inflammatory effects of dehydroxymethylepoxyquinomicin (DHMEQ), a nuclear factor-κB (NF-κB) inhibitor, on endotoxin-induced uveitis (EIU) in rats.

Methods: EIU was induced by a subcutaneous injection of lipopolysaccharide (LPS) in Lewis rats. DHMEQ was injected intraperitoneally concurrently with the LPS. Aqueous humor was collected 24 h after the LPS injection. Isolated peritoneal exudate cells (PECs) were exposed to LPS with or without DHMEQ to determine the production of TNF-α, IL-6, and MCP-1.

Results: DHMEQ significantly reduced the number of infiltrating cells, and the concentrations of proteins, TNF-α, and IL-6 in the aqueous humor. DHMEQ suppressed the production of TNF-α, IL-6, and MCP-1 from PECs. Immunochemistry revealed a reduction in the translocation of the NF-κB p65 into the nuclei in DHMEQ-exposed PECs.

Conclusions: The results indicate that DHMEQ has anti-inflammatory effects on EIU and may be a promising agent to treat intraocular inflammation.