Experimental Study of Aqueous Outflow Facility : Eye Perfusion Technique and Tissue Culture Methods

流出的房水被认为是沿着压力梯度穿过小梁网（TM），而不需要从TM的细胞泵送能量。然而，小梁外流途径可能构成不仅仅是一个被动的，结构性过滤器的出口的房水从eye.TM细胞（第2或第3次传代）被放置在稀疏培养6天，然后暴露于媒体测试中存在或不存在的过氧化氢酶和超氧化物歧化酶等的影响进行了检查流出设施和细胞形状的变化。在一个平行的实验中，观察细胞形状的变化与各种细胞骨架影响药物和细胞骨架（肌动蛋白），并通过凝胶电泳。为了进一步了解水流出的机制，我们还比较了流出设施的变化与张力和神经介导的睫状肌收缩。NaF使设施减少了50%。碘乙酰胺和碘乙酸在35 ℃下均增加了易变性。2，4-二硝基苯酚、KCN和2-脱氧葡萄糖对细胞凋亡无影响。茶碱和异丁基甲基黄嘌呤增加流出设施。然而，无论肾上腺素或异丙肾上腺素增加，流出设施，无论其浓度。肾上腺素或异丙肾上腺素（10 <-8>~ 10 μ <-6>M）对牛眼睫状肌张力无松弛作用，而茶碱和IBMX对卡巴胆碱收缩的牛眼睫状肌有明显的松弛作用，并抑制神经介导的收缩。流出组织的形态学改变（细胞形状或细胞附着）和诱导的睫状肌反应可能在流出设施中起重要作用。

Outflow aqueous humor has been considered to pass through the trabecular meshwork(TM)down a pressure gradient without requiring pumping energy from the cells of the TM. However, trabecular outflow pathways may constitute more than merely a passive, structural filter for egress of aqueous humor from the eye.TM cells(2nd or 3rd subculture) were placed cultured in sparse for six days and then exposed to media tested in both the presence or absence of either catalase and superoxide dismutase etc. The effects were examined by changes of outflow facility and cell shape. In a parallel experiment, cell shape change was observed with various cytoskeleton affecting drugs and by cell cytoskeleton (actin) and by gel electrophoresis.To further understand the aqueous outflow mechanism, we also compared changes in the outflow facility with the tone and nerve-mediated contraction of ciliary muscle. NaF reduced the facility as much as 50%. lodoacetamide and iodoacetate both increased facility at 35 C. 2, 4 dinitrophenol, KCN and 2-deoxyglucose did not produce any effect. Theophylline and isobutylmethylxantnine increased outflow facility. However, neither epinephrine nor isoproterenol increased the, outflow facility, regardless of their concentrations. Epinephrine or isoproterenol(10^<-8>-10^<-6> M)did not relax the tone of bovine ciliary muscle, whereas theopliylline and IBMX remarkably relaxed the muscle which had been contracted by carbachol and also inhibited the nerve-mediated contraction.These findings suggest that several substances in the aqueous humor may influence outflow facility. Morphological change(cell shape or cell attachment)of outflow tissues and the induced response of ciliary muscle may play a significant role on the outflow facility.