MacGregor L C, Rosecan L R, Laties A M, Matschinsky F M
J Biol Chem. 1986 Mar 25;261(9):4046-51.
Total lipid, sucrose, glucose, sorbitol and myo-inositol contents in individual layers from normal and alloxan-diabetic rabbit retinas were measured using gravimetric and enzymatic microtechniques. Pure samples of nine retinal layers were microdissected from freeze-dried retinal cryosections. The lipid content was measured by loss of weight after ethanol and hexane extraction. Retinal lipid varied 3-fold across the retina and was not influenced by diabetes. Sucrose, glucose, sorbitol, and myo-inositol were measured with fluorimetric microassays. Sucrose infused intravenously prior to sampling the tissue did not traverse the outer blood retinal barrier of the normal or the diabetic retina. In both normals and diabetics, glucose followed a diffusional curve, with outer and inner retinal concentrations, respectively, equal to choroidal blood and vitreal glucose concentrations. Sorbitol was elevated in all retinal layers of diabetic animals. The peak sorbitol concentrations, of 2 mmol/kg defatted dry weight in diabetics, were not high enough to be osmotically significant. Retinal myo-inositol, of the order found in brain tissue (10-30 mmol/kg defatted dry tissue), was decreased by 22-40% in all retinal layers of the diabetics. The results indicate that diabetes affects the metabolism of retinal structures independently of the small blood vessel disease that is the hallmark of diabetes of long duration. It is conceivable that primary intraretinal metabolic alterations cause, aggravate, or perpetuate the well known degenerative processes that occur in retinal blood vessels in diabetes.
采用重量法和酶促微量技术测定了正常和四氧嘧啶糖尿病兔视网膜各层中的总脂质、蔗糖、葡萄糖、山梨醇和肌醇含量。从冷冻干燥的视网膜冰冻切片中显微切割出9个视网膜层的纯样本。通过乙醇和己烷提取后的重量损失来测量脂质含量。视网膜脂质在整个视网膜中的变化幅度为3倍,不受糖尿病影响。蔗糖、葡萄糖、山梨醇和肌醇采用荧光微量分析法进行测定。在对组织取样之前静脉注射的蔗糖未穿过正常或糖尿病视网膜的外血视网膜屏障。在正常人和糖尿病患者中,葡萄糖均呈扩散曲线,视网膜外层和内层的浓度分别等于脉络膜血和玻璃体葡萄糖浓度。糖尿病动物的所有视网膜层中山梨醇均升高。糖尿病患者中山梨醇的峰值浓度为2 mmol/kg脱脂干重,不足以产生显著的渗透压作用。视网膜肌醇的含量与脑组织中的含量相当(10 - 30 mmol/kg脱脂干组织),在糖尿病患者的所有视网膜层中均降低了22 - 40%。结果表明,糖尿病独立于长期糖尿病的标志性小血管疾病影响视网膜结构的代谢。可以想象,视网膜内原发性代谢改变会导致、加重或延续糖尿病患者视网膜血管中发生的众所周知的退行性过程。