Xu Han-Mei, Wei Jin, Pan Li, Lin Hongying, Wang Weiqiang, Zhang Yihua, Shen Zilong
Department of Marine Pharmacy, College of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, Jiangsu Province, China; E-Mails:
Int J Mol Sci. 2010 Sep 15;11(9):3323-33. doi: 10.3390/ijms11093323.
(R,R) ZX-5 has been proven to have positive effects on choroidal blood flow without affecting the sclera and ciliary bodies in New Zealand white rabbits. This study was designed to investigate the mechanisms of (R,R) ZX-5 on improving the choroidal blood flow and promoting NO production. HUVECs (human umbilical vein endothelial cells) were used to determine the production of eNOS, p-eNOS, AKT and Erk1/2 by Western blot analysis. iNOS and eNOS mRNA levels were investigated by RT-PCR and the effect of (R,R) ZX-5 on NO production were determined by eNOS activity assay. We found (R,R) ZX-5 upregulated protein expression of eNOS and iNOS, increased NO production, and reduced ERK and Akt protein level. Therefore, (R,R) ZX-5 may promote the choroidal blood flow in New Zealand white rabbits without affecting the blood flow in the iris or ciliary bodies via increasing NO production. These results suggest that (R,R) ZX-5 may function to cure and prevent Age-related macular degeneration (AMD).
已证实(R,R) ZX - 5对新西兰白兔脉络膜血流有积极影响，且不影响巩膜和睫状体。本研究旨在探讨(R,R) ZX - 5改善脉络膜血流及促进一氧化氮（NO）生成的机制。采用人脐静脉内皮细胞（HUVECs），通过蛋白质免疫印迹分析来测定内皮型一氧化氮合酶（eNOS）、磷酸化eNOS（p - eNOS）、蛋白激酶B（AKT）和细胞外信号调节激酶1/2（Erk1/2）的生成情况。通过逆转录聚合酶链反应（RT - PCR）研究诱导型一氧化氮合酶（iNOS）和eNOS的信使核糖核酸（mRNA）水平，并通过eNOS活性测定来确定(R,R) ZX - 5对NO生成的影响。我们发现(R,R) ZX - 5上调了eNOS和iNOS的蛋白表达，增加了NO生成，并降低了ERK和Akt蛋白水平。因此，(R,R) ZX - 5可能通过增加NO生成来促进新西兰白兔的脉络膜血流，而不影响虹膜或睫状体的血流。这些结果表明，(R,R) ZX - 5可能具有治疗和预防年龄相关性黄斑变性（AMD）的作用。
