NOS3和SOD2在非糖尿病老年性白内障发生中的生物学关联及表达

Shamim Mushtaq, Meraj Zehra, A. Khan, Mehwish Ahmed, R. Ghani, N. Ahmed
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引用次数: 0

摘要

目的:探讨一氧化氮合酶(NOS)和超氧化物歧化酶(SOD)在非糖尿病人老年性白内障晶状体发病中的作用关系。方法:采用二维凝胶电泳(2-DE)方法对人白内障晶状体与正常晶状体(对照)的总溶解蛋白进行比较。采用基质辅助激光解吸/电离飞行时间质谱法(MALDI-TOF-MS)鉴定了不同丰度的蛋白质。Western blot检测NOS3和SOD2的表达变化。在STRING 8.3数据库中发现NOS3与SOD2和其他蛋白的进一步功能关联。结果:在2-DE图谱中,白内障蛋白和正常晶状体蛋白在pH 3 ~ 10范围内迁移,相对分子量为20 ~ 130 kDa。MALDI-TOF-MS检测到两个强度不同的蛋白点分别为NOS3和SOD2。Western blot分析显示,白内障组织中NOS3高表达,正常晶状体组织中SOD2高表达。串相互作用网络显示NOS3和SOD2之间的强相互作用具有高可信度,这有助于表征可能是白内障发病原因的功能异常。结论:本研究为白内障发生的机理评价和预防提供了新的途径。然而,需要大规模的研究来评估这种相互作用对人类白内障临床结果的影响。
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Biological Association and Expressions of NOS3 & SOD2 in Non-Diabetic Senile Cataractogenesis
Aim: To evaluate the functional relationship between the nitric oxide synthase (NOS) and superoxide dismutase (SOD) enzymes in the pathogenesis of human senile cataract lenses of non-diabetic patients. Methods: Total solubilized proteins from human cataract lens were compared with normal lens (control) by 2-Dimenstional gel electrophoresis (2-DE). Proteins with different abundances were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Western blot analysis was used to verify the changes in expression of NOS3 and SOD2. A further functional association of NOS3 with SOD2 and other proteins was seen by STRING 8.3 databases. Results: In the 2-DE maps, the cataract and normal lens proteins migrated in the region of pH 3 - 10 with a relative molecular weight of 20 - 130 kDa. Approximately two protein spots with differential intensity were detected as NOS3 and SOD2 using MALDI-TOF-MS. Western blot analysis showed high expression of NOS3 in cataract and SOD2 in normal lens samples. String interaction network revealed strong interactions between NOS3 and SOD2 at high confidence score, which is helpful in characterization of functional abnormalities that may be a causative factor in the pathogenesis of cataract. Conclusion: This study will offer new avenues for mechanistic evaluation and future prevention of cataractogensis. However, large scale studies will be required to evaluate the effect of this interaction on the clinical outcome in human cataract.
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