Wu Renyi, Yin Jinfu, Yao Ke, Haefliger Ivan
Eye Center of the 2nd Affiliated Hospital, Medical College of Zhejiang University, Hangzhou, China.
Mol Vis. 2007 Apr 2;13:493-6.
To investigate the mRNA and protein expression of nitric oxide synthase (NOS) in human ciliary bodies in vitro. The effect of the adenylcyclase activator forskolin and/or the alpha2-adrenergic agonist brimonidine (an ocular hypotensive agent that inhibits aqueous humor formation) on NOS mRNA or protein expression was also studied.
Frozen human ciliary bodies obtained from local eye bank were thawed and incubated with 0.1 mM forskolin for 24 h in the absence or in the presence of 100 muM brimonidine. The mRNA and protein expression of three NOS isoforms (neuronal NOS or nNOS, inducible NOS or iNOS, endothelial NOS or eNOS) were assessed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis, respectively.
mRNA and protein expression of three NOS isoforms were detected in human ciliary bodies. Forskolin significantly up-regulated the mRNA and protein expression of nNOS, but not that of iNOS or of eNOS. In the presence of brimonidine, the forskolin-induced up-regulation of nNOS mRNA or protein expression was significantly inhibited.
In human ciliary body (where aqueous humor is produced), brimonidine inhibits the up-regulation of nNOS expression induced by forskolin.
研究一氧化氮合酶（NOS）在人睫状体中的mRNA和蛋白表达。同时研究腺苷酸环化酶激活剂福司可林和/或α2 - 肾上腺素能激动剂溴莫尼定（一种抑制房水生成的降眼压药物）对NOS mRNA或蛋白表达的影响。
从当地眼库获取的冷冻人睫状体解冻后，在不存在或存在100 μM溴莫尼定的情况下，与0.1 mM福司可林孵育24小时。分别通过逆转录 - 聚合酶链反应（RT - PCR）和蛋白质印迹分析评估三种NOS同工型（神经元型NOS或nNOS、诱导型NOS或iNOS、内皮型NOS或eNOS）的mRNA和蛋白表达。
在人睫状体中检测到三种NOS同工型的mRNA和蛋白表达。福司可林显著上调nNOS的mRNA和蛋白表达，但不影响iNOS或eNOS的表达。在存在溴莫尼定的情况下，福司可林诱导的nNOS mRNA或蛋白表达上调受到显著抑制。
在人睫状体（房水产生的部位）中，溴莫尼定抑制福司可林诱导的nNOS表达上调。
