眼科 ›› 2013, Vol. 22 ›› Issue (6): 383-388.

• 论著 • 上一篇    下一篇

β趋化因子受体CCR1在遗传性视网膜变性小鼠视网膜中的表达

曾惠阳   陈星星  唐胜建  卢清君  刘谦   

  1. 100730 首都医科大学附属北京同仁医院 北京同仁眼科中心 北京市眼科学与视觉科学重点实验室(曾惠阳、卢清君、刘谦);261053 潍坊医学院整形外科研究所(陈星星、 唐胜建)
  • 收稿日期:2013-06-26 出版日期:2013-11-25 发布日期:2013-12-10
  • 通讯作者: 曾惠阳,Email: zhydr@hotmail.com E-mail:zhydr@hotmail.com
  • 基金资助:

    国家自然科学基金(81100675);北京市自然科学基金(7102034)

Expression of β-chemokine receptor CCR1 in the retina of inherited retinal dystrophy mice

ZENG Hui-yang1, CHEN Xing-xing2,TANG Sheng-jian2, LU Qing-jun1, LIU Qian1   

  1. 1. Beijing Tongren Eye Center, Bejing Key Laboratory of Ophthalmology and Visual Science, Beijing Tongren Hospital, Capital Medical University, Beijng 100730, China; 2. Plastic Surgery Research Center, Weifang Medical University, Weifang 261053, China
  • Received:2013-06-26 Online:2013-11-25 Published:2013-12-10
  • Contact: ZENG Hui-yang, Email: zhydr@hotmail.com E-mail:zhydr@hotmail.com

摘要: 目的 探讨β趋化因子主要的受体之一 β趋化因子受体1(CCR1)在视网膜变性模型小鼠(rd小鼠)的视网膜变性过程中的表达及其在感光细胞凋亡中的病理作用。设计 实验研究。研究对象 出生后8、10、12、14、16及18天的rd小鼠各10只(共60只)及同龄C57BL/6N对照小鼠各10只(共60只)。方法 小鼠断颈处死,取双眼眼球制作冰冻切片或分离新鲜视网膜组织。逆转录多聚酶链反应(RT-PCR)测定各鼠龄rd小鼠及对照鼠视网膜CCR1mRNA的表达水平。免疫组织化学法观察CCR1蛋白在各鼠龄rd小鼠视网膜的定位表达。CCR1在感光细胞及凋亡细胞中的表达由免疫荧光双标法确定。主要指标 视网膜CCR1mRNA及蛋白的表达、CCR1的细胞定位及与凋亡细胞的关系。结果 CCR1mRNA在对照组及各鼠龄rd小鼠视网膜中均有表达,但在出生后12、14天rd小鼠视网膜中表达明显升高(光密度值分别为0.986±0.17和1.152±0.22,P=0.010和0.008)。CCR1阳性染色细胞开始出现于出生后8天的rd视网膜外核层,并于12及14天达到高峰。对照组视网膜外核层无CCR1阳性染色。CCR1与视紫红质、CD11b或TUNEL染色免疫荧光双标结果显示,CCR1表达于感光细胞而非小胶质细胞中,部分CCR1表达于凋亡的感光细胞中。结论 在rd小鼠视网膜中CCR1表达于感光细胞并随其变性程度加重表达升高。CCR1的活化可能在rd小鼠感光细胞凋亡中发挥作用。

关键词: &beta, 趋化因子受体, 小胶质细胞, 感光细胞凋亡, rd小鼠

Abstract: Objective To investigate the expression of C-C chemokine receptor 1(CCR1), a major chemokine receptor for β-chemokines, in the ratina of  rd (retinal degeneration) mice and further explore its role in the photoreceptor degeneration. Design Experimental study. Partrcipants Sixty rd mice at postnatal days (P) 8, 10, 12, 14, 16 and 18 day (10 mice each) and 60 C57BL/6N control mice with the matcholde age (10 mice each). Methods The expression levels of CCR1mRNA in the whole control  rd mice were determined by RT-PCR assay. Location of CCR1 in the retina were studied by immunohistochemical analysis. Expression of CCR1 in the photoreceptor cells and apoptotic cells was determined by double labeling. Main Outcome Measures Expression of CCR1mRNA and CCR1 protein, cellular location of CCR1 and its relation to the apoptotic cells.  Results Expression of CCR1mRNA was noted in both control and rd mice retinas at each age group, but was markedly increased in the rd mice retinas at P12 and P14 day. CCR1-positive cells started to emerge in the rd mice retina at P8 day and reached a peak at P12 and P14 day. Double labeling of CCR1 with rhodopsin, CD11b or TUNEL staining showed expression of CCR1 in the photoreceptor cells, rather in the microglial cells. Partial CCR1 expression was observed in some of the apoptotic photoreceptor cells. Conclusion Expression of CCR1 in the photoreceptor cells was increased with progression of retinal degeneration in rd mice. Activation of CCR1 may play a role in the photoreceptor apoptosis.

Key words: β-chemokine receptor, microglia, photoreceptor apoptosis, rd mice