缓激肽刺激体外培养的视网膜色素上皮细胞炎症反应的作用机制
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国家自然基金面上项目(No.81470648)


Mechanism of bradykinin on inflammations of retinal pigment epithelium cells
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National Natural Science Foundation of China(No. 81470648)

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    摘要:

    目的:探讨缓激肽刺激体外培养的视网膜色素上皮(retinal pigment epithelium,RPE)细胞炎症反应的作用机制。

    方法:体外培养ARPE-19细胞,100nmol/L缓激肽(bradykinin,BK)刺激24h后,光镜下观察细胞形态变化,细胞免疫荧光检测BK受体定位; 共聚焦显微镜检测BK及其拮抗剂作用下Ca2+变化; Western blot检测对照组与100nmol/L BK处理24h后(BK组)COX-1、COX-2、eNOS、iNOS蛋白的表达量。

    结果:BK刺激后,ARPE-19细胞形态无明显变化; ARPE-19细胞可检测到激肽B1、B2受体; 与对照组相比,BK组Ca2+浓度显著升高; B1R拮抗组及B2R拮抗组的Ca2+浓度较BK组升高幅度降低,B1R及B2R拮抗组Ca2+浓度较对照组无明显变化; BK作用ARPE-19细胞后,COX-2及iNOS蛋白含量显著增加(P<0.001)。

    结论:BK通过与B1R及B2R结合促进体外培养的ARPE细胞COX-2及iNOS表达增加。

    Abstract:

    AIM: To investigate mechanism of bradykinin(BK)on inflammations of retinal pigment epithelium(RPE)cells.

    METHODS: ARPE-19 cells were cultured in vitro, stimulated by 100nM BK for 24h. Cell morphology changes were observed by microscope, and BK receptor localization was detected through cell immunofluorescence. Changes of Ca2+ in BK and BR antagonist stimuli were detected by laser scanning confocal microscopy. The expressions of COX-1, COX-2, eNOS and iNOS protein in control group and BK group were detected by Western Blot.

    RESULTS: After the stimulation of BK, there was no significant changes of ARPE-19 cells in morphology. Kinin B1 receptors(B1R)and B2 receptors(B2R)could be detected in ARPE-19 cells. Compared with control group, Ca2+ concentrations significantly increased in BK group; in B1R antagonist group and B2R antagonist group Ca2+ concentrations increased less than BK group; B1R and B2R antagonist group showed no obvious changes in Ca2+ concentrations. Compared with control group, COX-2 and iNOS protein concentrations were significantly increased in BK group(P<0.001).

    CONCLUSION: BK induces the increasing expression of COX-2 and iNOS in the cultured ARPE cells through binding with either B1R or B2R.

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蔡雯婷,任成达,刘晴雨,等.缓激肽刺激体外培养的视网膜色素上皮细胞炎症反应的作用机制.国际眼科杂志, 2016,16(8):1430-1434.

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  • 收稿日期:2016-03-01
  • 最后修改日期:2016-07-05
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  • 在线发布日期: 2016-07-26
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