Neurturin-deficient mice develop dry eye and keratoconjunctivitis sicca
Neurturin-deficient mice develop dry eye and keratoconjunctivitis sicca
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DOI：
10.1167/iovs.02-1319
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发表时间：
2003-10-01
影响因子：
4.4
通讯作者：
Pflugfelder, SC
中科院分区：
文献类型：
--
作者：
Song, XJ;Li, DQ;Pflugfelder, SC
目的。Neurturin已被确定为副交感神经元的神经营养因子。神经蛋白缺陷（NRTN-/-）小鼠泪腺副交感神经支配有缺陷。本研究对NRTN-/-小鼠的泪液功能和眼表表型进行了评价。以6周龄~ 4月龄NRTN-/-小鼠25只，NRTN+/+小鼠17只，采用尾基因组DNA PCR检测。连续测定泪液生成、泪液荧光素清除率和角膜感觉。角膜对AlexaFluor右旋糖酐（AFD; Molecular Probes, Eugene， OR）的渗透性通过荧光测定法在485 nm激发和530 mn发射下测量。在pas染色切片上评估组织学。免疫荧光染色法检测黏液蛋白和HLAⅱ（IA）抗原。ELISA法检测泪液il -1 β，酶谱法检测泪液基质金属蛋白酶(MMP)-9。采用半定量rt - pcr方法分析角膜上皮组织中基因的表达。与年龄匹配的NRTN+/+小鼠相比，NRTN+/+小鼠的泪液生成、泪液荧光素清除率和角膜感觉显著降低，而角膜对AFD的通透性显著增加。NRTN-/-小鼠结膜内muc4和- 5ac粘蛋白活性和杯状细胞密度均低于NRTN+/+小鼠（P < 0.001）。NRTN-/-小鼠角膜上皮muc1和- 4mrna的表达减少。NRTN-/-小鼠结膜上皮细胞IA抗原阳性的数量明显多于NRTN+/+小鼠。与NRTN+/+小鼠相比，NRTN-/-小鼠泪液中IL-1 β和MMP-9浓度以及角膜上皮中IL-1 β、tnf - α、巨噬细胞炎症蛋白(MIP)-2、细胞因子诱导的中性粒细胞趋化剂（KC）和MMP-9 mRNA的表达均显著升高。神经蛋白缺陷小鼠表现出表型改变和眼表炎症，类似于人类角膜结膜炎。该模型支持功能性眼表-中枢神经系统-泪腺感觉-自主神经网络在维持眼表健康和体内平衡中的重要性。
PURPOSE. Neurturin has been identified as a neurotrophic factor for parasympathetic neurons. Neurturin-deficient (NRTN-/-) mice have defective parasympathetic innervation of their lacrimal glands. This study was conducted to evaluate tear function and ocular surface phenotype in NRTN-/- mice.METHODS. Determined by tail genomic DNA PCR, 25 NRTN-/- mice and 17 neurturin-normal (NRTN+/+) mice aged 6 weeks to 4 months were evaluated. Aqueous tear production, tear fluorescein clearance and corneal sensation were serially measured. Corneal permeability to AlexaFluor dextran (AFD; Molecular Probes, Eugene, OR) was measured by a fluorometric assay at 485 nm excitation and 530 mn emission. Histology was evaluated in PAS-stained sections. Mucin and HLA class II (IA) antigen were assessed by immunofluorescent staining. Tear IL-1beta was measured by ELISA, and tear matrix metalloproteinase (MMP)-9 by zymography. Gene expression in the corneal epithelia was analyzed by serniquantitative RT-PCR.RESULTS. In comparison to that in age-matched NRTN+/+ mice, aqueous tear production, tear fluorescein clearance, and corneal sensation were significantly reduced in NRTN+/+ mice, whereas corneal permeability to AFD was significantly increased. Inummoreactive MUC-4 and -5AC mucin and goblet cell density (P < 0.001) in the conjunctiva of NRTN-/- mice were lower than in NRTN+/+ mice. The expression of MUC-1 and -4 mRNA by the corneal epithelium was reduced in NRTN-/- mice. There were a significantly greater number of IA antigen-positive conjunctival epithelial cells in NRTN-/- mice than NRTN+/+ mice. Tear fluid IL-1 beta and MMP-9 concentrations and the expression of IL-1 beta, TNF-alpha, macrophage inflammatory protein (MIP)-2, cytokine-induced neutrophil chemoattractant (KC), and MMP-9 mRNA by the corneal epithelia were significantly increased in NRTN-/- mice, compared with NRTN+/+ mice.CONCLUSIONS. Neurturin-deficient mice show phenotypic changes and ocular surface inflammation that mimic human keratoconjunctivitis sicca. This model supports the importance of a functional ocular surface-central nervous system-lacrimal gland sensory-autonomic neural network in maintaining ocular surface health and homeostasis.