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  

Mechanism of bradykinin on inflammations of retinal pigment epithelium cells

作　　者：蔡雯婷[1] 任成达[1] 刘晴雨[1] 危清泉 杜亚茹[1] 王倩怡[1] 刘俊伶[1] 何梦梅 于靖[1] 

机构地区：[1]同济大学附属第十人民医院眼科,中国上海市200072

出　　处：《国际眼科杂志》2016年第8期1430-1434,共5页International Eye Science

基　　金：国家自然基金面上项目(No.81470648)~~

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。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. Com pared with control group,Ca^（2＋）concentrations significantly increased in BK group; in B1R antagonist group and B2R antagonist group Ca^（2＋）concentrations increased less than BK group; B1R and B2 R antagonist group showed no obvious changes in Ca^（2＋） concentrations. Com pared with control group, COX-2and 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.

关 键 词：缓激肽 增生性视网膜病变 环氧化酶 一氧化氮合酶 

分 类 号：R774.1[医药卫生—眼科]