Prostaglandin E receptor subtype EP3 in conjunctival epithelium regulates late-phase reaction of experimental allergic conjunctivitis
Prostaglandin E receptor subtype EP3 in conjunctival epithelium regulates late-phase reaction of experimental allergic conjunctivitis
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DOI：
10.1016/j.jaci.2008.09.044
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发表时间：
2009-02-01
影响因子：
14.2
通讯作者：
Kinoshita, Shigeru
中科院分区：
文献类型：
--
作者：
Ueta, Mayumi;Matsuoka, Toshiyuki;Kinoshita, Shigeru
背景：我们之前已经证明前列腺素E-2 (PGE(2))-EP3通路负调控小鼠过敏性哮喘模型中的过敏反应。目的：探讨PGE(2)-EP3通路是否也调控小鼠实验性变应性结膜炎（EAC）的发生。方法：采用RTPCR和免疫组化方法检测野生型小鼠EP3的表达，采用5-溴-4-氯-3-吲哚- β - d-半乳糖苷染色法检测EP3基因缺失小鼠（Ptger3（-/-）小鼠）在EP3基因位点携带r -半乳糖苷酶基因的表达。用短豚草花粉（RW）免疫小鼠诱导EAC，再用短豚草花粉滴眼液刺激，观察结膜内嗜酸性粒细胞的浸润和eotaxin-1 mRNA的表达。小鼠也在诱导期局部应用ep3选择性激动剂。采用定量RT-PCR检测cox和前列腺素E合成酶的表达，ELISA检测眼睑组织中PGE(2)的产生。结果：EP3在眼表结膜上皮中组成性表达。与野生型小鼠相比，Ptger3（-/-）小鼠在RW刺激后结膜内嗜酸性粒细胞浸润显著增加。同样，在Ptger3（-/-）小鼠中观察到显著更高的eotaxin-1 mRNA表达。相反，用ep3选择性激动剂治疗野生型小鼠可导致嗜酸性粒细胞浸润显著减少，而在Ptger3（-/-）小鼠中则被钝化。RW刺激后，眼睑内COX-2和前列腺素E合成酶的表达上调，PGE(2)含量升高。结论：PGE(2)作用于结膜上皮EP3，下调EAC的进展。[J] .中华过敏症杂志，2009;23(3):466- 471。
Background: We previously demonstrated that the prostaglandin E-2 (PGE(2))-EP3 pathway negatively regulates allergic reactions in a murine allergic asthma model.Objectives: We investigated whether the PGE(2)-EP3 pathway also regulates the development of murine experimental allergic conjunctivitis (EAC).Methods: The expression of EP3 was examined by means of RTPCR and immunohistochemistry in wild-type mice, as well as by means of 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside staining in mice deficient in EP3 (Ptger3(-/-) mice) carrying the R-galactosidase gene at the EP3 gene locus. EAC was induced by immunization of mice with short ragweed pollen (RW), followed by challenge with eye drops of RW, and eosinophil infiltration and eotaxin-1 mRNA expression in the conjunctiva were examined. Mice were also treated with a topical application of an EP3-selective agonist during the elicitation phase. Quantitative RT-PCR was used to detect expression of COXs and prostaglandin E synthases, and ELISA was used to measure PGE(2) production in the eyelid.Results: EP3 was constitutively expressed in conjunctival epithelium on the ocular surface. Ptger3(-/-) mice demonstrated significantly increased eosinophil infiltration in conjunctiva after RW challenge compared with wild-type mice. Consistently, significantly higher expression of eotaxin-1 mRNA was observed in Ptger3(-/-) mice. Conversely, treatment of wild-type mice with an EP3-selective agonist resulted in a significant decrease in eosinophil infiltration, which was blunted in Ptger3(-/-) mice. Expression of COX-2 and prostaglandin E synthases was upregulated and PGE(2) content was increased in the eyelids after RW challenge.Conclusion: These data suggest that PGE(2) acts on EP3 in conjunctival epithelium and downregulates the progression of EAC. (J Allergy Clin Immunol 2009;123:466-71.)