解剖学报

解剖学报 ›› 2015, Vol. 46 ›› Issue (2): 265-269.doi: 10.16098/j.issn.0529-1356.2015.02.020

• 组织学胚胎学发育生物学 • 上一篇    下一篇

G蛋白耦联受体30对去卵巢小鼠子宫中toll样受体4分布与表达的影响

刘红改 王璐 代盈盈 于得水 赵慧英*   

  1. 西北农林科技大学动物医学院， 陕西 杨凌 712100
  • 收稿日期:2014-06-20 修回日期:2014-11-15 出版日期:2015-04-06 发布日期:2015-04-06
  • 通讯作者: 赵慧英 E-mail:zhaohuiying@nwsuaf.edu.cn
  • 基金资助:

    抗病转基因牛新品种培育重点专项

Effect of G protein-coupled receptor 30 on the distribution and expression of toll-like receptor 4 in the uterus of ovariectomized mice

LIU Hong-gai WANG Lu DAI Ying-ying YU De-shui ZHAO Hui-ying*   

  1. College of Veterinary Medicine，Northwest A&F University，Shanxi Yangling 712100，China
  • Received:2014-06-20 Revised:2014-11-15 Online:2015-04-06 Published:2015-04-06
  • Contact: ZHAO Hui-ying E-mail:zhaohuiying@nwsuaf.edu.cn
  • Supported by:

    specific research on disease-resistant transgenic cattle breeding

摘要：

目的 探讨雌激素膜性受体G蛋白耦联受体30(GPR30)在子宫免疫中的作用。方法 将80只小鼠分为假手术（sham）组、卵巢摘除（OVX）组、OVX注射0.1nmol、0.5nmol和2.5nmol GPR30激动剂 G1组及OVX小鼠注射0.1nmol、0.5nmol和2.5nmol GPR30拮抗剂 G15组，分别采用免疫组织化学SP法及实时定量PCR技术，对子宫中toll样受体4（TLR4）的分布及TLR4 mRNA的表达量进行研究。 结果 TLR4免疫反应阳性产物主要分布在小鼠子宫的内膜上皮、腺上皮及间质细胞；G1高浓度组中TLR4的免疫组织化学阳性着色强度显著低于其他组(P<0.01)，去卵巢小鼠给予GPR30拮抗剂G15后，TLR4的相对表达量随G15浓度的升高而升高，各浓度组间TLR4的相对表达量显著性变化差异同OVX+G1组。各组小鼠子宫中TLR4 mRNA的表达量变化与TLR4相对表达量变化一致。 结论 GPR30可下调子宫中TLR4的表达，并可能介导雌激素参与子宫的局部免疫功能的调节。

关键词: G蛋白耦联受体30, 子宫, Toll样受体4, 免疫组织化学SP法, 实时定量聚合酶链反应, 去卵巢小鼠

Abstract:

Objective To investigate the role of G protein-coupled receptor 30（GPR30）on the uterine immunity. Methods A total of 80 mice were used in this study. Immunochemistry SP method and fluorescent real-time quantitative PCR technology were used to study the distribution of toll-like receptor（TLR4）and the expression of TLR mRNA in sham group, ovariectomized (OVX) group, OVX+0.1nmol G1 group, OVX+0.5nmol G1 group, OVX+2.5nmol G1 group, OVX+0.1nmol G15 group, OVX+0.5nmol G15 group, OVX+2.5nmol G15 group. Results TLR4 was mostly localized to the cell membrane and cytoplasm of the luminal, glandular epitheliumcells and mesenchymal cells. The intensities of the immunohistochemical positive staining for TLR4 were significantly lower in G1 high concentration group than those in other groups(P<0.01). After administrating the antogonist G15 to the OVX mice, the relative expression of TLR4 was increased with G15 concentration increasing. The significant difference on the relative expression of TLR4 in OVX+G15 group was coincidence with OVX+G1 group. The result of fluorescent realtime quantitative PCR showed that the expression level change of TLR4 mRNA in uterus of mice between groups was consistent with the relative expression of TLR4. Conclusion These results indicate that GPR30 can inhibit the expression of TLR4 in the uterus，and that GPR30 may participate in regulating the uterus local immune function.

Key words: G protein-coupled receptor 30, Uterus, Toll-like receptor 4, Immunochemical SP method, Real-time PCR, Ovariectomized mouse