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β趋化因子通过小胶质细胞活化引起小鼠感光细胞凋亡的实验研究

刘谦  陈艳  王沿强  曾惠阳   

  1. 100005 首都医科大学附属北京同仁医院  北京同仁眼科中心  北京市眼科研究所;眼科学与视觉科学北京市重点实验室(刘谦、曾惠阳);100029 中国科学院北京基因研究所(陈艳、王沿强)
  • 收稿日期:2017-03-02 出版日期:2017-07-25 发布日期:2017-07-24
  • 通讯作者: 曾惠阳,Email: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)

The effect of β-chemokine on microglia and photoreceptor cells of mice in vitro

LIU Qian1, CHEN Yan2, WANG Yan-qiang2, ZENG Hui-yang1.   

  1. 1. Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University; Beijing Ophthalmology & Visual Sciences Key Laboratory, Beijing 100005, China; 2. Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing 100029, China.
  • Received:2017-03-02 Online:2017-07-25 Published:2017-07-24
  • Contact: ZENG Hui-yang, Email: zhydr@hotmail.com

摘要:

 目的 观察β趋化因子对小鼠小胶质细胞的活化及诱导感光细胞凋亡的作用。设计 实验性研究。研究对象 β趋化因子、小鼠BV-2小胶质细胞系及661w感光细胞系。方法 将β趋化因子(RANTES、MIP-1α及MIP-1β)加入到BV-2细胞中,观察BV-2细胞活化反应。然后将被激活的BV-2细胞培养液上清加入661w细胞中,观察661w的凋亡情况。或将β趋化因子直接加入到661w细胞中,观察其凋亡情况。事先加入β趋化因子抑制剂met-RANTES后,重复上述操作。主要指标 钙内流、ROS及NO的产生、TUNEL阳性感光细胞百分比。结果 β趋化因子加入BV-2细胞后,细胞内钙流曲线明显上升、细胞外ROS及NO产生较对照组均明显增高(P<0.01);将激活后的BV-2细胞上清加入到661w细胞中,后者凋亡率增加30%。β趋化因子直接加入661w细胞中,上述指标无明显变化。在加入β趋化因子之前先加入其抑制剂met-RANTES,BV-2细胞的ROS产生较未加抑制剂组明显降低(P<0.01)、NO产生较未加抑制剂组降低(P=0.0187),661w细胞凋亡明显减少。结论 β趋化因子可活化小鼠小胶质细胞导致感光细胞凋亡。β趋化因子抑制剂可抑制遗传性视网膜变性小鼠小胶质细胞活化,保护感光细胞。(眼科, 2017, 26:270-275)
 

关键词: &beta, 趋化因子;小胶质细胞;感光细胞;BV-2细胞系;661w细胞系

Abstract:

Objective  To observe the activation of microglia and the photoreceptor cell apoptosis of mice induced by β-chemokines in vitro.  Design  Experimental study. Participants β-chemokine, BV-2 cells of mice, 661w cells of mice. Methods Activation of BV-2 cells were studied when β-chemokines (RANTES, MIP-1αandMIP-1β) were added. The cell death of 661w cells was observed when the supernatant of the activated BV-2 cells was added into the 661w cells. The 661w cell apoptosis was also studied when the β-chemokines were directly added. Met-RANTES (the inhibitor of β-chemokine) was added before hand and above markers were studied. Main Outcome Measures Calcium influx, ROS, NO, TUNEL positive cells. Results The calcium influxesas well as release of ROS, NO by the BV-2 cells was increased (P<0.01) after adding β-chemokine.The 661w cell death was elevated by 30% after adding the supernatant of activated microglia. Above markers can be inhibited by the β-chemokine inhibitors(ROS P<0.01, NO P=0.0187).There was no change when adding β-chemokine into 661w cells directly.  Conclusions β-chemokines can activate microglia, which induce photoreceptor cell death by release of ROS or NO. Photoreceptor degeneration of RP mice may be protected by inhibition of β-chemokine inhibitors. (Ophthalmol CHN, 2017, 26: 270-275)

Key words: β-chemokine, microglia, photoreceptor; , BV-2 cells, 661wcells