Lim Seul-Ki, Park Min-Jung, Jung Ho-Kyoung, Park Ah-Yeon, Kim Dong-Il, Kim Jong-Chun, Bae Chun-Sik, Kim Kye-Yeop, Yoon Kyoung-Chul, Han Ho Jae, Park Soo Hyun
Bio-therapy Human Resources Center, Department of Veterinary Physiology, College of Veterinary Medicine, Chonnam National University, Gwangju 500-757, Republic of Korea.
Life Sci. 2008 Dec 5;83(23-24):761-70. doi: 10.1016/j.lfs.2008.09.014. Epub 2008 Sep 30.
We were to examine the effect of bradykinin (BK) in the regulation of glutamate transporter and its related signaling molecules in a human retinal pigment epithelial (ARPE) cells, which are important cells to support retina.
d-[2,3-(3)H]-aspartate uptake, western immunoblotting, reverse transcription polymerase chain reaction, [(3)H]-arachidonic acid release, and siRNA transfection techniques were used.
BK stimulated glutamate uptake as well as the mRNA expression of excitatory amino acid transporter 4 (EAAT4) and excitatory amino acid carrier 1 (EAAC1), which was blocked by treatment with bradykinin 1 receptor (B1R) and bradykinin 2 receptor (B2R) siRNA, suggesting the role of B1R and B2R in this process. The BK-induced stimulation of glutamate uptake was also blocked by [des-Arg(10)]-HOE 140, a B1R antagonist, and HOE 140, a B2R antagonist, as well as by the tyrosine kinase inhibitors genistein and herbimycin A. In addition, the BK-induced stimulation of glutamate uptake was blocked by treatment with the phospholipase A(2) inhibitors mepacrine and AACOCF(3), the cyclooxygenase (COX) inhibitor indomethacin, and the COX-2 inhibitor Dup 697. Furthermore, the BK-induced increase in COX-2 expression was blocked by the PI-3 kinase inhibitors wortmannin and LY294002, Akt inhibitor, and the protein kinase C (PKC) inhibitors staurosporine and bisindolylmaleimide I, suggesting the role of PI-3 kinase and PKC in this process. BK stimulated Akt activation and the translocation of PKC activation via the activation of B1R and B2R.
BK stimulates glutamate uptake through a PKC-Akt-COX-2 signaling cascade in ARPE cells.
我们旨在研究缓激肽（BK）对人视网膜色素上皮（ARPE）细胞中谷氨酸转运体及其相关信号分子的调节作用，ARPE细胞是支持视网膜的重要细胞。
采用d-[2,3-(3)H]-天冬氨酸摄取、蛋白质免疫印迹、逆转录聚合酶链反应、[(3)H]-花生四烯酸释放和小干扰RNA（siRNA）转染技术。
BK刺激谷氨酸摄取以及兴奋性氨基酸转运体4（EAAT4）和兴奋性氨基酸载体1（EAAC1）的mRNA表达，缓激肽1受体（B1R）和缓激肽2受体（B2R）的siRNA处理可阻断这一作用，提示B1R和B2R在此过程中的作用。BK诱导的谷氨酸摄取刺激也被B1R拮抗剂[去-精氨酸(10)]-HOE 140、B2R拮抗剂HOE 140以及酪氨酸激酶抑制剂染料木黄酮和除莠霉素A阻断。此外，BK诱导的谷氨酸摄取刺激被磷脂酶A(2)抑制剂米帕林和AACOCF(3)、环氧化酶（COX）抑制剂吲哚美辛以及COX-2抑制剂Dup 697阻断。此外，BK诱导的COX-2表达增加被磷脂酰肌醇-3激酶（PI-3激酶）抑制剂渥曼青霉素和LY294002、Akt抑制剂以及蛋白激酶C（PKC）抑制剂星形孢菌素和双吲哚基马来酰亚胺I阻断，提示PI-3激酶和PKC在此过程中的作用。BK通过激活B1R和B2R刺激Akt活化和PKC活化的转位。
BK通过ARPE细胞中的PKC-Akt-COX-2信号级联刺激谷氨酸摄取。
