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

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Ca~...

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Bibliographic Details
Published in国际眼科杂志 Vol. 16; no. 8; pp. 1430 - 1434
Main Author 蔡雯婷 任成达 刘睛雨 危清泉 杜亚茹 王倩怡 刘俊伶 何梦梅 于靖
Format Journal Article
LanguageChinese
Published 同济大学附属第十人民医院眼科, 中国上海市,200072 2016
Subjects
一氧化氮合酶
增生性视网膜病变
环氧化酶
缓激肽
proliferative vitreoretinopathy
bradykinin
cyclooxygenase
增生性视网膜病变
缓激肽
一氧化氮合酶
nitric oxide synthase
环氧化酶
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Summary: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Ca~（2＋）浓度显著升高;B1R拮抗组及B2R拮抗组的Ca~（2＋）浓度较BK组升高幅度降低,B1R及B2R拮抗组Ca~（2＋）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。
Bibliography:bradykinin; proliferative vitreoretinopathy; cyclooxygenase; nitric oxide synthase
Wen-Ting Cai,Cheng-Da Ren,Qing-Yu Liu,Qing-Quan Wei,Ya-Ru Du,Qian-Yi Wang,Jun-Ling Liu,Meng-Mei He,Jing Yu （Department of Ophthalmology, the Tenth People＇s Hospital of Tongji University, Shanghai 200072, China）
61-1419/R
AIM： To investigate mechanism of bradykinin（ BK） on inflam m ations of retinal pigm ent epithelium（ RPE） cells.METHODS： ARPE-19 cells were cultured in vitro,stim ulated by 100 n MBK for 24 h. Cell m orphology changes were observed by m icroscope,and BK receptor localization was detected through cell im m unofluorescence. Changes of Ca2＋in BK and BR antagonist stim uli were detected by laser scanning confocal m icroscopy. The expressions of COX-1,COX-2,eNOS and i NOS 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 m orphology. Kinin B1receptors（ B1R） and B2 receptors（ B2R） could be detected in ARPE-19 cells
ISSN:1672-5123
DOI:10.3980/j.issn.1672-5123.2016.8.07