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。

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.