PROTEIN-COMPOSITION OF HUMAN AQUEOUS-HUMOR - SDS-PAGE ANALYSIS OF SURGICAL AND POST-MORTEM SAMPLES
PROTEIN-COMPOSITION OF HUMAN AQUEOUS-HUMOR - SDS-PAGE ANALYSIS OF SURGICAL AND POST-MORTEM SAMPLES
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DOI：
10.1016/0014-4835(89)90025-0
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发表时间：
1989-01-01
影响因子：
3.4
通讯作者：
TRIPATHI, BJ
中科院分区：
文献类型：
--
作者：
TRIPATHI, RC;MILLARD, CB;TRIPATHI, BJ
我们分析了人类房水的蛋白质组成。通过穿刺术从25只择期白内障手术的人眼（年龄范围64-92岁）和20只年龄匹配的死后1.5至18小时内的眼睛中获得样本。对单个样品进行总蛋白测定，并通过定性SDS-PAGE将多肽分离成高、中、低分子量范围，然后进行银染。临床样品在总蛋白值方面显示出显著的一致性（平均值± 0.001）。SEM：12.4 ±。2.0 mg/100 ml），并且在银染蛋白质的谱中没有可检测的变化。存在12个主要蛋白质级分，其表观分子量为140、80（双峰）、67、60（双峰）、35、27、25、17、14.6和9 kDa。初步分析表明，17 kDa条带含有类似碱性成纤维细胞生长因子的分子。另外两个来自手术时获得血液/眼房水的患者的眼房水样品，死后样品显示出总蛋白含量的更大变异性（56.1 ± 0.01）。11.6 mg/100 ml）和增加的高分子量和低分子量蛋白质级分的数量。考虑到临床参数（包括眼部和全身用药、全身性疾病、手术前用药、麻醉和总血清蛋白值）的广泛差异，仔细抽取的手术样本的蛋白质谱的相似性最为显著。我们的研究结果表明，在接受择期白内障手术的患者中，人房水中主要蛋白质的水平不受临床参数的广泛个体差异的影响。我们将这一发现归因于在收集房水样本时所采取的护理措施。
We analyzed the protein composition of human aqueous humor. Samples were obtained by paracentesis from 25 human eyes (age range 64-92 years) at elective cataract surgery, and from 20 age-matched post-mortem eyes within 1.5 to 18 hr after death. Individual samples were assayed for total protein, and the polypeptides were separated by qualitative SDS-PAGE into high-, medium- and low-molecular-weight ranges and then silver-stained. The clinical samples showed a remarkable consistency in the total protein values (mean .+-. SEM: 12.4 .+-. 2.0 mg per 100 ml) and no detectable variations in the profiles of the silver-stained proteins. Twelve major protein fractions, with apparent molecular weights of 140, 80 (doublet), 67, 60 (doublet), 35, 27, 25, 17, 14.6 and 9 kDa, were present. A preliminary analysis showed that the 17 kDa band contained a molecule resembling basic fibroblast growth factor. Two additional samples of aqueous humor from patients whose blood/aqueous humor obtained at surgery, the post-mortem samples exhibited a greater variability in total protein content (56.1 .+-. 11.6 mg per 100 ml) and an increased number of high- and low-molecular-weight protein fractions. In view of wide differences in the clinical parameters, including ocular and systemic medications, systemic illness, surgical premedications, anesthesia and total serum protein values, the similarity in the protein profiles of the carefully drawn surgical samples is most remarkable. Our results indicate indicate that, in patients who underwent elective cataract surgery, the levels of major proteins in human aqueous humor are not affected by wide individual variations in the clinical parameters. We attribute this finding to the care taken in the collection of aqueous humor samples.