调节性T细胞在实验性自身免疫性葡萄膜炎中的作用

目的探讨CD4^＋调节性T细胞（Treg）在实验性自身免疫性葡萄膜炎（EAU）病程进展中的作用。方法通过IRBP161-180和完全弗氏佐剂皮下免疫,在B10RⅢ小鼠诱导产生EAU,对眼内炎症行临床检查并对其严重程度予以评分。通过免疫磁珠分选EAU恢复期小鼠脾脏CD4^＋T细胞、CD8^＋T细胞、CD4^＋CD25^＋T细胞和CD4^＋CD25^-T细胞。通过流式细胞术分析脾脏CD4^＋CD25^＋Foxp3^＋T细胞的比例。通过体内过继转移评价Treg的抑制效应。结果小鼠诱导EAU后产生眼内炎症反应。将EAU恢复期小鼠脾细胞、脾CD4＋T细胞或脾CD4^＋CD25^＋T细胞过继转移入正常小鼠继而皮下免疫诱导EAU,眼底检查显示受者小鼠眼内炎症程度明显降低。相反,正常小鼠脾细胞、EAU恢复期脾CD8^＋T细胞或CD4^＋CD25^-T细胞过继转移无此效应。流式细胞分析显示随着EAU的发展,脾内CD4^＋CD25^＋T细胞比例和CD4^＋CD25^＋Foxp3^＋T细胞比例逐步升高,并在恢复期维持较高水平。结论 CD4^＋Treg随着EAU的病程进展数量增多,它们可能参与EAU的自发缓解。     

实验性自身免疫性葡萄膜炎T淋巴细胞亚群与Treg细胞的表达研究

目的　探讨实验性自身免疫性葡萄膜炎（ｅｘｐｅｒｉｍｅｎｔａｌａｕｔｏｉｍｍｕｎｅｕｖｅｉｔｉｓ,ＥＡＵ）新西兰兔模型外周血单个核细胞（ｐｅ-ｒｉｐｈｅｒａｌｂｌｏｏｄｍｏｎｏｎｕｃｌｅａｒｃｅｌｌｓ,ＰＢＭＣ）Ｔ淋巴细胞亚群的变化及其与疾病特征的相关性。方法　选取健康成年雄性新西兰大白兔４０只作为动物模型,用２０ｇ·Ｌ－１牛血清蛋白（ｂｏｖｉｎｅｓｅｒｕｍａｌｂｕｍｉｎ,ＢＳＡ）静脉注射和玻璃体内注射法建立ＥＡＵ模型,并观察模型眼炎症反应及ＰＢＭＣＴ淋巴细胞亚群特征。结果　ＣＤ４＋Ｔ细胞在ＥＡＵ兔的外周血中比例明显增加,且随时间的进展呈进行性增加,并与炎症反应呈正相关,相同的趋势可见于ＣＤ４＋／ＣＤ８＋Ｔ细胞比例。此外,Ｔｒｅｇ细胞趋势与ＣＤ４＋Ｔ细胞及ＣＤ４＋Ｔ／ＣＤ８＋Ｔ细胞比例相反,与炎症反应呈负相关。结论　ＣＤ４＋Ｔ细胞的优势性选择性克隆增殖及Ｔｒｅｇ细胞的降低有可能引发免疫功能紊乱并导致针对自身组织的免疫应答失去有效的负性调控,导致ＥＡＵ的发生及进行性加重。     

Th1、Th17细胞相关因子在实验性自身免疫性葡萄膜炎大鼠中的表达及作用

目的　探讨辅助性Ｔ细胞（Ｔｈｅｌｐｅｒｃｅｌｌｓ,Ｔｈ）１、Ｔｈ１７细胞相关因子在实验性自身免疫性葡萄膜炎（ｅｘｐｅｒｉｍｅｎｔａｌａｕｔｏｉｍｍｕｎｅｕｖｅｉｔｉｓ,ＥＡＵ）中的表达及作用。方法　取清洁级纯系健康雌性Ｌｅｗｉｓ大鼠４０只随机分为ＥＡＵ组（３２只）和对照组（８只）,ＥＡＵ组用光感受器间维生素Ａ类结合蛋白诱导大鼠ＥＡＵ模型,进行临床症状评分,免疫组织化学方法检测造模后视网膜内干扰素（ｉｎｆｅｒｆｅｒｏｎ,ＩＦＮ）-γ、诱导型一氧化氮合酶（ｉｎｄｕｃｉｂｌｅｎｉｔｒｉｃｏｘｉｄｅｓｙｎｔｈａｓｅ,ｉＮＯＳ）、白细胞介素（ｉｎｔｅｒｌｅｕｋｉｎｅ,ＩＬ）-１７、ＩＬ-６的表达。ＥＬＩＳＡ法对比分析房水中各细胞因子的变化情况。结果　ＥＡＵ模型建立成功;造模后第１４天,视网膜损害以外层为主,视网膜内有大量炎细胞浸润,从而导致视网膜内结构紊乱,同时在视网膜的视锥、视杆细胞层和神经节细胞层ｉＮＯＳ、ＩＬ-１７、ＩＬ-６、ＩＦＮ-γ表达,细胞阳性率分别为２９％、４８％、５２％、７３％。在ＥＡＵ发病过程中,ＩＬ-６于造模后第７天迅速升高,第１０天达到高峰;ＩＬ-１７于造模后第１４天达到最高值,变化趋势与炎症进程一致;ＩＦＮ-γ在炎症后期仍有升高,于造模后第１６天达到最高值;ＩＬ-６、ＩＬ-１７、ＩＦＮ-γ与对照组相比,各时间点表达差异均有统计学意义（均为Ｐ＜０．０５）。ｉＮＯＳ在炎症进程中表达有所增加,但与对照组相比,各时间点的表达差异均无统计学意义（均为Ｐ＞０．０５）。结论　Ｔｈ１、Ｔｈ１７细胞相关因子调节网络共同参与实验性自身免疫性葡萄膜炎的发生发展。     

实验性自身免疫性葡萄膜炎大鼠的发病特点

背景 Lewis大鼠是建立实验性自身免疫性葡萄膜炎（EAU）动物模型的常用种系,对其发病特点,尤其是EAU大鼠眼部超微结构改变的研究尚未见报道. 目的 观察EAU大鼠的发病体征及其眼部超微结构的改变. 方法 选取SPF级6～8周龄雌性Lewis大鼠18只,采用随机数字表法随机分为对照组6只和模型组12只.模型组大鼠后肢足底、两侧腹壁和躯干上皮下各注射含有光感受器间维生素A类结合蛋白（IRBP,1177-1191）和结核菌素（TB）的完全弗氏佐剂（CFA）乳化液,对照组大鼠不作处理.注射后观察两组大鼠饮食、体温和活动情况,每天裂隙灯显微镜下观察大鼠眼部炎症表现,于免疫后12d获取大鼠眼组织标本,对虹膜、睫状体和视网膜进行常规组织病理学检查,并分别在扫描电子显微镜和透射电子显微镜下观察虹膜、睫状体和视网膜的超微结构改变.结果 模型组大鼠免疫后采食量为（190.00± 18.03）g,明显少于对照组的（285.33±28.02）g,差异有统计学意义（t=4.955,P=0.012）;模型组和对照组大鼠的饮水量分别为（241.67±18.56）ml和（289.67±18.18）ml,差异有统计学意义（t=3.201,P=0.033）;模型组大鼠体温升高,精神倦怠.裂隙灯显微镜下观察发现,模型组大鼠免疫后6d出现虹膜充血、前房积脓和瞳孔膜闭,免疫后12d眼部炎症最严重,炎症评分为（3.83±0.41）分,而对照组大鼠眼前节未见异常.组织病理学检查发现,模型组大鼠前房、虹膜、睫状体组织和玻璃体腔内均可见大量淋巴细胞、中性粒细胞和单核巨噬细胞浸润.扫描电子显微镜下可见模型组大鼠虹膜肌纤维粗细不均,睫状体上皮表面粗糙及RPE细胞绒毛疏松.透射电子显微镜下观察可见,模型组大鼠虹膜单核巨噬细胞浸润,睫状体上皮细胞膜褶皱蓬松及排列紊乱,视网膜Müller细胞中有髓样小体,RPE细胞中有线粒体空泡出现,而对照组大鼠虹膜、睫状体及     

β-arrestin1在实验性自身免疫性葡萄膜炎中的表达

目的:初步探讨β-arrestin1在实验性自身免疫性葡萄膜炎（experimental autoimmune uveitis,EAU）中的表达。 方法：成功建立EAU动物模型后,通过免疫组织化学和RT-PCR的方法检测β-arrestin1在EAU小鼠脾脏细胞中的表达。 结果：免疫组织化学和RT-PCR均提示β-arrestin1在EAU小鼠脾脏细胞中的表达较正常小鼠增高。 结论：β-arrestin1可能在EAU的发生发展中起着重要作用。     

间充质干细胞对大鼠实验性自身免疫性葡萄膜炎视网膜中白细胞介素17表达的影响

背景 研究显示Th17细胞是实验性自身免疫性葡萄膜炎(EAU)发病的重要致炎细胞群,而白细胞介素17( IL-17)作为Th17细胞的标志性因子参与了EAU的发生与发展.间充质干细胞(MSCs)作为一种免疫调节剂在多种自身免疫性疾病中对Th17具有抑制作用. 目的 研究MSCs对EAU的治疗作用以及对IL-17在大鼠视网膜表达的影响.方法 从10只SPF级Wistar大鼠股骨骨髓中分离、培养MSCs并进行传代,第3～5代细胞用于实验.54只SPF级6～8周龄Lewis大鼠按随机数字表法分为实验组48只和正常对照组6只.光感受器间维牛素A类结合蛋白(IRBP)与含有结核杆菌素HR37a的2.5 g/L弗氏完全佐剂(CFA)等体积混合后行实验组大鼠单后足垫皮下注射建立EAU动物模型,然后按照分层随机原则分为模型对照组和MSCs治疗组,每组24只大鼠.MSCs治疗组于造模的同时行1 ml MSCs尾静脉注射(5×106个／ml),连续注射3d,模型对照组以同法注射等体积PBS.注射后每日在裂隙灯显微镜下观察大鼠眼部炎症反应,并根据Caspi临床分级进行评分.分别于造模后9、12、15、20 d随机选取模型对照组和MSCs治疗组各6只大鼠制备视网膜切片,采用组织病理学方法观察大鼠视网膜形态学改变和炎症反应,参照Caspi组织学分级法进行评分.采用免疫组织化学染色法检测造模后9、12、15、20d大鼠视网膜中IL-17的表达.结果 培养的细胞呈梭形生长,漩涡状排列,经流式细胞术鉴定CD29、CD44表达阳性,CD45、CD34表达阴性.MSCs治疗组造模后9、12、15、20 d眼前节炎症反应评分均低于模型对照组(U=2.815,P=0.005;U=2.768,P=0.006;U=2.900,P=0.004;U=2.855,P=0.004),视网膜组织学检测炎症评分均低于模型对照组,差异均有统计学意义( U=2.345,P=0.019:U=2.559,P=0.011;U=2.166,P=0.030;U=2.373,P=0.018).免疫组织化学染色显示,MSCs治疗组造模后9、12、15、20d大鼠视网膜IL-17蛋白阳性表达的平均吸光度(A)值分别为26.47±5.68、77.78±9.65、47.02±6.68和26.59±5.94,明显低于模型对照组的45.34±4.63、105.95±10.74、64.11±9.76和37.02±6.51,差异均有统计学意义(t=6.305,P=0.000:t=4.799,P=0.001:t=3.540,P=0.005;t=2.900,P=0.016).结论 MSCs能减轻EAU的程度并抑制EAU的进展,降低了1L-17在眼组织中的表达.     

实验性自身免疫性视网膜葡萄膜炎的研究

实验性自身免疫性视网膜葡萄膜炎动物模型的建立，对于开展葡萄膜炎的基础及临床研究有重要意义，它是第一个用于眼内炎症机制基础与临床研究可靠的动物模型，对于ＥＡＵ的研究涉及到免疫学。     

c-Myc抑制剂10058-F4对实验性自身免疫性葡萄膜炎小鼠模型Th17细胞比例及细胞因子表达的影响

目的　探讨c-Myc抑制剂10058-F4对实验性自身免疫性葡萄膜炎（EAU）小鼠模型光感受器间维生素A类结合蛋白1-20（IRBP_1-20）特异性辅助性T细胞17（Th17）细胞比例及细胞因子表达的影响。方法　取健康C57BL/6J小鼠（8~10周龄）6只,采用随机数字表法将小鼠分为正常对照组、EAU组,每组3只。EAU组小鼠腹腔内注射350 ng百日咳毒素,单后足和脊柱两侧皮下注射含150 μg IRBP_1-20、0.8 mg结核分枝杆菌H37RA和弗氏不完全佐剂的乳化剂以诱导EAU模型;正常对照组小鼠不给予特殊干预。免疫后13 d,行间接检眼镜检查并通过小动物视网膜成像系统可视化检测小鼠眼底炎性病灶,HE染色观察视网膜组织病理学改变,实时荧光定量PCR（qRT-PCR）检测T细胞和眼球组织中c-Myc mRNA的表达。将EAU模型小鼠T细胞分为对照组和10058-F4组,10058-F4组T细胞加入50 μmol·L^-1 10058-F4,对照组T细胞不给予药物干预。培养6 h后,洗去药物,将两组T细胞与IRBP_1-20（10 mg·L^-1）及骨髓来源树突细胞共培养,同时加入20 μg·L^-1 白细胞介素（IL）-23（Th17细胞诱导条件）。采用qRT-PCR检测c-Myc、维甲酸相关核孤儿受体γt（RORγt）、IL-17A、IL-17F和粒细胞-巨噬细胞集落刺激因子（GM-CSF） mRNA相对表达量,ELISA检测共培养细胞上清液中IL-17含量,流式细胞仪检测共培养细胞中Th17细胞比例。结果　本研究EAU组小鼠模型建立成功。qRT-PCR检测结果显示,EAU组小鼠T细胞、眼球组织中c-Myc mRNA相对表达量分别为1.85±0.37、2.31±0.47,较正常对照组（1.01±0.02、0.99±0.05）均明显升高,差异均有统计学意义（P=0.017、0.008）;10058-F4组T细胞c-Myc mRNA相对表达量为0.57±0.03,明显低于对照组（1.04±0.02）,差异有统计学意义（P<0.001）。ELISA检测结果显示,10058-F4组共培养细胞上清液中IL-17含量明显低于对照组,差异有统计学意义（P=0.025）。流式细胞仪检测结果显示,10058-F4组共培养细胞中Th17细胞比例为（17.97±0.91）%,明显低于对照组［（22.50±0.90）%］,差异有统计学意义（P=0.004）。qRT-PCR检测结果显示,10058-F4组T细胞RORγt、IL-17A、IL-17F、GM-CSF mRNA相对表达量分别为0.52±0.11、0.51±0.06、0.58±0.15、0.75±0.03,明显低于对照组（1.01±0.01、1.00±0.01、1.02±0.02、1.01±0.02）,差异均有统计学意义（均为P<0.05）。结论　c-Myc抑制剂10058-F4可显著降低T细胞中c-Myc表达水平,抑制Th17细胞特异性转录因子RORγt表达及效应细胞因子IL-17A、IL-17F、GM-CSF的产生,负向调控IRBP_1-20特异性Th17细胞免疫反应。     

雷帕霉素对大鼠实验性自身免疫性葡萄膜炎的治疗作用

背景雷帕霉素不仅具有抗菌作用,而且是一种良好的免疫抑制剂,可用于多种自身免疫性疾病的治疗．对实验性自身免疫性葡萄膜炎（EAU）的治疗作用是目前研究的热点之一。目的研究雷帕霉素对EAU的治疗作用,并观察雷帕霉素对EAU各免疫细胞群炎性因子表达的影响。方法25只Lewis大鼠采用随机数字表法分为EAU组（20只）和正常对照组（5只）。光感受器间维生素A类结合蛋白（IRBP）R16肽段与完全氟氏佐剂充分乳化后于Lewis大鼠后足皮下注射以建立EAU模型,EAU模型鼠再按分层随机的原则分为模型对照组和雷帕霉素组,每组10只大鼠。雷帕霉素组造模后即应用O．2mg／（kg·d）雷帕霉素（0．4m1）腹腔内连续注射7d,模型对照组及正常对照组大鼠采用等体积的生理盐水进行腹腔内注射。造模后第4天开始每日裂隙灯下观察大鼠EAU的症状,造模后第14天制备大鼠视网膜切片,以苏木精-伊红染色法进行组织病理学观察,参照Caspi的标准对EAU症状及组织病理学分级进行评分。应用免疫组织化学染色法检测各组大鼠视网膜中炎性因子干扰素-γ（IFN-γ）、白细胞介素-17（IL-17）的表达情况。结果造模后6d模型对照组大鼠EAU炎症评分逐渐升高,12d达到高峰,然后逐渐下降。雷帕霉素组大鼠EAU炎症评分变化呈现相同的趋势,但各时间点EAU炎症评分均明显低于模型对照组,差异均有统计学意义（P〈0．01）。视网膜组织病理学研究表明,模型对照组大鼠视网膜结构紊乱,大量炎性细胞浸润,组织病理学评分为3．30±0．48,而雷帕霉素组视网膜结构接近正常,组织学评分为0．90±0．45,差异有统计学意义（t=16．541,P〈0．01）。雷帕霉素组IFN-γ、IL-17在大鼠视网膜中的表达量（A值）分别为21．16±4．23和49．86±6．59,明显低于模型对照组的62．14±7．32和124．85±6．33,差异均有统计学意义（q=33．334、q=56．923,P〈0．01）。结论雷帕霉素通过抑制EAU视网膜中IFN-γ、IL-17等炎性因子的表达而对EAU发挥治疗作用,其机制可能是通过抑制Th1、Th17细胞群来实现的。     

γδ T细胞在实验性自身免疫性葡萄膜炎小鼠脾脏中的动态表达及作用机制

背景 以往研究证明葡萄膜炎的发病机制与γδ T细胞相关,γδ T细胞在实验性自身免疫性葡萄膜炎(EAU)中的作用尚不完全清楚. 目的 研究γδ T细胞在EAU小鼠脾脏中的动态变化,探讨γδ T细胞在EAU病程进展中的作用机制.方法 应用随机数字表法将45只C57 BL/6(B6)小鼠随机分为正常对照组6只和模型组39只.用抗原肽光感受器间维生素A类结合蛋白1-20(IRBP1-20)及完全弗氏佐剂(CFA)的乳化液在B6小鼠的足垫、尾根部及躯干部均匀注射6个点,用Genesis-D动物眼部照相机观察并记录正常小鼠及免疫后不同时间点(4、8、12、16、20、24、28、32和36 d)EAU小鼠的发病情况,参照Thurau的评分标准进行炎症评分.颈椎脱臼法处死小鼠后摘取右侧眼球,切片后行组织病理学评分.同时在相同时间点分离模型小鼠脾脏淋巴细胞,流式细胞仪检测γδ T细胞数量和激活状态.免疫磁珠分选γδ T细胞,并用流式细胞仪检测其细胞内表达白细胞介素-17A(IL-17A)的变化.向EAU小鼠回输激活的γδ T细胞,观察炎症变化. 结果 经IRBP1-20及CFA乳化液免疫后小鼠于第12天可见轻度葡萄膜炎症,炎症反应在免疫后第16 ～20 d达峰,至第28天炎症明显减轻.组织病理学观察发现,免疫小鼠视网膜外核层皱褶,玻璃体、视网膜全层炎性细胞浸润及内界膜层增厚.造模后不同时间点的炎症症状评分和病理学炎症评分的总体比较,差异均有统计学意义(F=51.399,P=0.000;F=47.342,P=0.000).流式细胞仪检测发现,EAU发病高峰期小鼠的脾脏中γδ T细胞数量增加,造模后16 d和20 d分别为(5.67±0.49)％和(5.78±0.55)％,与造模前的(1.53±0.14)％比较,差异均有统计学意义(均P＜0.05),且呈活化状态.EAU发病高峰期小鼠的脾脏中γδ T细胞分泌的IL-17A明显增多,造模后16d和20 d分别为(13.40±0.50)％和(17.80±2.37)％,与正常对照小鼠的(1.53±0.19)％比较,差异均有统计学意义(P=0.000、0.001).体内回输激活的γδ T细胞后EAU炎症加重,炎症症状评分为1.00(1.00,2.00),明显高于未回输激活的γδ T细胞的0.75(0.05,1.00),二者比较差异有统计学意义(Z=27.00,P=0.03).结论 EAU中γδ T细胞比例在炎症的高峰期明显升高,且呈激活状态;活化性Yδ T细胞在EAU模型中发挥的免疫调节作用可能是通过分泌IL-17A进行的.     

Effect of Gallium Nitrate on Experimental Autoimmune Uveitis

Gallium nitrate (GN) has been shown to inhibit T cell-mediated inflammatory disease. The purpose of our study was to test the effect of gallium nitrate (GN) on experimental autoimmune uveitis (EAU). Experimental autoimmune uveitis was induced in male Lewis rats immunized with retinal S-antigen. Rats received subcutaneous injections of GN or saline one day prior to immunization and 1, 4, 7, 10, 13, 16, and 19 days after immunization. Ocular inflammation was graded clinically and histologically by masked observers, and in vitro assays of cell-mediated and humoral immunity were performed. GN significantly inhibited the development of EAU graded clinically (P=0.001) and histologically (P=0.002). Treatment with GN also resulted in a small (30–41%) decrease in the lymphocyte responses to retinal S-Antigen and a small (12–37%) reduction in antibody production to S-antigen. These data show that GN suppresses the development of EAU, and inhibits both lymphocyte proliferative responses to antigen and antibody production.     

Antimetabolite Drugs Exhibit Distinctive Immunomodulatory Mechanisms and Effects on the Intestinal Microbiota in Experimental Autoimmune Uveitis

PurposeThe purpose of this study was to investigate the effect of antimetabolite drugs on T-cell responses and intestinal microbial composition in autoimmune uveitis.MethodsExperimental autoimmune uveitis (EAU) was induced in C57BL/6J mice treated with 0.00625 mg/mL methotrexate (MTX) or 0.625 mg/mL mycophenolate mofetil (MMF) in drinking water for 4 weeks prior to immunization and 2 weeks thereafter. The effector T cell (Teff) and regulatory T cell (Treg) populations were identified using flow cytometry. The 16S rRNA gene sequencing was applied for gut microbiome characterization. DESeq2 analysis was used to discriminate relative abundances of taxa and PLS-DA to integrate cytometric and microbiome data between groups.ResultsBoth MTX and MMF abrogated uveitis in EAU without clinical signs of toxicity as compared to water-fed controls. MTX reduced Teff and Treg expansion in peripheral tissues and eyes. MTX decreased alpha diversity, increased Akkermansia, and reduced Lachnoclostridium abundances. Conversely, MMF enhanced Tregs in the mesenteric lymph node and the eyes. In parallel, MMF increased the gut alpha diversity, including an increased abundance of Lachnospiraceae NK4A136 group and a decreased abundance of Lachnospiraceae UCG-001. A significant congruent correlation among intestinal microbial changes, T-cell responses, and clinical scores was observed for both antimetabolites.ConclusionsAlthough MTX and MMF both abrogated uveitis in EAU, they showed different effects on T-cell subsets and the intestinal bacterial composition. This work indicates unique immunomodulation by each drug and is the first to demonstrate potential microbiota-related mechanisms.     

白藜芦醇对内毒素诱导的大鼠葡萄膜炎的治疗作用

目的：探讨眼局部使用白藜芦醇（Res）对内毒素诱导的大鼠葡萄膜炎的治疗效果及相关机制。方法：实验研究。将36 只雄性Wistar大鼠按随机数字表法分为空白组、模型组、0.25% Res治疗组、0.5%Res治疗组、1% Res治疗组和0.1%地塞米松（Dex）治疗组,每组6只。将大肠杆菌细胞壁脂多糖（LPS）溶于无菌的0.9%氯化钠溶液配成1 mg/ml的LPS溶液,并注射入模型组和治疗组大鼠双后足垫（每只注射200 μg）,空白组注射等量0.9%氯化钠溶液。各治疗组于LPS诱导前后分别给予相应浓度的 Res和Dex 10 μl点双眼,每2 小时1 次,注射LPS前后各6 次。空白组和模型组给予等量的0.9%氯化钠溶液点眼。观察大鼠眼部炎症反应、临床表现评分,注射LPS 24 h后测房水肿瘤坏死因子-α （TNF-α）和白细胞介素-6（IL-6）浓度、眼球病理切片HE染色以及免疫组织化学检测虹膜睫状体p38丝裂原活化蛋白激酶（MAPK）和核转录因子-κB（NF-κB）p65 等,探讨与分析Res的治疗效果。采用单因素方差分析、Mann-Whitney U检验进行数据处理。结果：模型组于LPS注射后4 h可见虹膜血管扩张充血,之后炎症反应逐渐加重,24 h时可见前房大量点状渗出、瞳孔区纤维渗出及晶状体前囊膜渗出物沉着,模型组24 h临床评分为4.3 ± 1.2,1% Res组为2.0 ± 0.6,1% Res组葡萄膜炎反应明显轻于模型组（P < 0.05）;1% Res组房水中TNF-α和IL-6的浓度与模型组相比均明显降低（P < 0.05）;1% Res组眼部组织病理学改变较模型组轻（P < 0.05）;1% Res组虹膜睫状体p38 MAPK和NF-κB p65 的核转位阳性细胞率较模型组均明显降低（P < 0.05）。结论：1% Res局部点眼能有效减轻内毒素诱导的大鼠自身免疫性葡萄膜炎的临床症状,其机制可能与抑制MAPK和NF-κB信号通路有关。     

Human Embryonic Stem Cell-Derived Mesenchymal Stromal Cells Decrease the Development of Severe Experimental Autoimmune Uveitis in B10.RIII Mice

Purpose: We investigated the effect of exogenously administered human embryonic stem cell-derived mesenchymal stromal cells (hESC-MSCs) in experimental autoimmune uveitis (EAU) in B10.RIII mice, a murine model of severe uveitis.

Methods: B10.RIII mice were immunized with an uveitogenic peptide, and intraperitoneal injections of 5 million hESC-MSCs per animal were given on the same day. Behavioral light sensitivity assays, histological evaluation, cytokine production, and regulatory T cells were analyzed at the peak of the disease.

Results: Histological and behavioral evidence demonstrated that early systemic treatment with hESC-MSCs decreases the development of severe EAU in B10.RIII mice. hESC-MSCs suppress Th17 and upregulate Th1 and Th2 responses as well as IL-2 and GM-CSF in splenocytes from hESC-MSC-treated mice.

Conclusions: MSCs that originate from hESC decrease the development of severe EAU in B10.RIII mice, likely through systemic immune modulation. Further investigation is needed to determine any potential effect on active EAU.     

Differential Effect of Macrophage Depletion on Two Forms of Experimental Uveitis Evoked by Pigment Epithelial Membrane Protein (EAPU), and by Melanin-Protein (EMIU)

The purpose of the present study was to clinically and histologically investigate the influence of macrophage depletion on the development of experimental autoimmune pigment epithelial membrane protein-induced uveitis (EAPU), and experimental melanin-protein induced uveitis (EMIU) in the Lewis rat. EAPU is mainly characterized by pigment epitheliitis. Posterior mononuclear cell accumulations enclose and destroy the retinal pigment epithelium (RPE). In EMIU the inflammation is specifically localized in the uvea.EAPU was induced by immunization with RPE membrane protein, and EMIU was evoked by immunization with purified choroidal melanin. Systemic treatment with dichloromethylene diphosphonate (Cl₂MDP)-containing liposomes just before the expected beginning of the clinical signs of EAPU (at day 7 and 9 after immunization) resulted in a considerable delay of the uveitis process. In the treated animals the typical plaque shaped cell accumulations (containing many macrophages) along the RPE were lacking. Two weeks after the treatment, severe rebound EAPU developed. Local treatment by subconjunctival liposome injections did not exert any effect on EAPU. In EMIU, macrophage depletion by systemic treatment did not noticeably influence the clinical and histological development of the inflammation.Systemic treatment at the peak stage of EAPU (at day 12 and 14 after immunization) resulted in the rapid disappearance of the clinical signs of uveitis. Vitreous and anterior chamber cells were virtually absent two days later. This situation remained unchanged until the experiment was terminated two weeks later. Already deposited cell accumulations along the RPE did not regress but stopped their progression.Hematogenous macrophages thus appear to play a crucial role in the development of EAPU but the effect of early macrophage depletion on EAPU appeared to be temporary due to blood repopulation. A possible explanation for the differential influence of macrophage depletion on EAPU and EMIU is discussed, and is based on differences in immunopathogenesis.     

短期高浓度葡萄糖摄入对小鼠实验性自身免疫性葡萄膜炎发展的作用

目的 探索短期高浓度葡萄糖摄入对小鼠实验性自身免疫性葡萄膜炎(EAU)发展的促进作用.方法 选取健康SPF级6～8周龄小鼠38只,随机分为模型对照组(EAU组)和高糖模型组(GLU组),每组19只小鼠.GLU组小鼠从造模前14 d开始给予200 g·L-1葡萄糖溶液饮水直至处死,EAU组小鼠给予正常饮水,同时保证EAU...     

Effect of Liposome—Encapsulated Total Alkaloid of Harmaline on Rabbit Lens Epithelial Cells：Experimental Study on the Prevention of Posterior Capsule Opacification

Purpose: To investigate whether liposome encapsulated total alkaloid of Harmaline (TAH) as a therapeutic agent is beneficial to prevention of posterior capsular opacifi-cation (PCO).Methods: Liposome-encapsulated TAH was prepared by modified freeze-thawing method. 0. 1ml of liposome-encapsulated TAH (0. 2mg/ml) was injected into the capsular bag during extracapsular lens extraction (ECLE) of each eye in total 10 rabbit eyes. Blank liposome or balance salt solution (BSS) was used as control. Slit-lamp examination and histopathological examination was used to evaluated capsule opacifica-tion. Intraocular pressure (IOP) , density and morphology of corneal endothelia cells, the amplitude and latency of b wave of ERG were measured.Results: The inflammatory response was mild both in TAH treated and the control group. PCO formation occurred in the control group 2 weeks postoperatively, but the posterior capsule was clear in TAH treated eyes. 4 weeks and 8 weeks after operation, PCO occurred both in TAH treated