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.
References
[1]
Foster, A. and Resnikoff, S. (2005) The Impact of Vision 2020 on Global Blindness. Eye, 19, 1133-1135. http://dx.doi.org/10.1038/sj.eye.6701973
[2]
Beatty, S., Koh, H., Phil, M., Henson, D. and Boulton, M. (2000) The Role of Oxidative Stress in the Pathogenesis of Age-Related Macular Degeneration. Survey of Ophthalmology, 3, 115-134. http://dx.doi.org/10.1016/S0039-6257(00)00140-5
[3]
Mukesh, B.N., Le, A., Dimitrov, P.N., Ahmed, S., Taylor, H.R. and McCarty, C.A. (2006) Development of Cataract and Associated Risk Factors: The Visual Impairment Project. Archives of Ophthalmology, 124, 79-85. http://dx.doi.org/10.1001/archopht.124.1.79
[4]
Ashok, B.T. and Ali, R. (1999) The Aging Paradox: Free Radical Theory of Aging. Experimental Gerontology, 34, 293-303. http://dx.doi.org/10.1016/S0531-5565(99)00005-4
[5]
Qianqian, Y., Yong, Y., Zhaodong, C., Yonghui, T, Jun, S. and Yuzheng, H. (2015) Differential Protein Expression between Type 1 Diabetic Cataract and Age-Related Cataract Patients. Folia Biologica (Praha), 61, 74-80.
[6]
Zhou, H.Y., Yan, H., Wang, L.L., Yan, W.J., Shui, Y.B. and Beebe, D.C. (2015) Quantitative Proteomics Analysis by iTRAQ in Human Nuclear Cataracts of Different Ages and Normal Lens Nuclei. Proteomics: Clinical Applications, 9, 776-786.
http://dx.doi.org/10.1002/prca.201400061
[7]
Harman, D. (1956) Aging: A Theory Based on Free Radical and Radiation Chemistry. The Journals of Gerontology, 11, 298-300. http://dx.doi.org/10.1093/geronj/11.3.298
[8]
Chang, D., Zhang, X., Rong, S., Sha, Q., Liu, P., Han, T. and Pan, H. (2013) Serum Antioxidative Enzymes Levels and Oxidative Stress Products in Age-Related Cataract Patients. Oxidative Medicine and Cellular Longevity, 587826. http://dx.doi.org/10.1155/2013/587826
[9]
Øsnes-Ringen, O., Azqueta, A.O., Moe, M.C., Zetterstrøm, C., Röger, M., Nicolaissen, B. and Collins, A.R. (2013) DNA Damage in Lens Epithelium of Cataract Patients in Vivo and ex Vivo. Acta Ophthalmologica, 91, 652-656.
http://dx.doi.org/10.1111/j.1755-3768.2012.02500.x
[10]
Katta, A.V., Katkam, R.V. and Geetha, H. (2013) Lipid Peroxidation and the Total Antioxidant Status in the Pathogenesis of Age Related and Diabetic Cataracts: A Study on the Lens and Blood. Journal of Clinical and Diagnostic Research, 7, 978-981.
http://dx.doi.org/10.7860/jcdr/2013/4937.3099
[11]
Kaur, J., Kukreja, S., Kaur, A., Malhotra, N. and Kaur, R. (2012) The Oxidative Stress in Cataract Patients. Journal of Clinical and Diagnostic Research, 6, 1629-1632.
http://dx.doi.org/10.7860/jcdr/2012/4856.2626
[12]
Sorte, K., Sune, P., Bhake, A., Shivkumar, V.B., Gangane, N. and Basak, A. (2011) Quantitative Assessment of DNA Damage Directly in Lens Epithelial Cells from Senile Cataract Patients. Molecular Vision, 17, 1-6.
[13]
Beebe, D.C., Holekamp, N.M. and Shui, Y.B. (2010) Oxidative Damage and the Prevention of Age-Related Cataracts. Ophthalmic Research, 44, 155-165.
http://dx.doi.org/10.1159/000316481
[14]
Selin, J.Z., Lindblad, B.E., Rautiainen, S., Michaëlsson, K., Morgenstern, R., Bottai, M., Basu, S. and Wolk, A. (2014) Are Increased Levels of Systemic Oxidative Stress and Inflammation Associated with Age-Related Cataract? Antioxidants & Redox Signaling, 21, 700- 704. http://dx.doi.org/10.1089/ars.2014.5853
[15]
Cekic, S., Zlatanovic, G., Cvetkovic, T. and Petrovic, B. (2010) Oxidative Stress in Cataractogenesis. Bosnian Journal of Basic Medical Sciences, 10, 265-269.
[16]
Nagai, N., Liu, Y., Fukuhata, T. and Ito, Y. (2006) Inhibitors of Inducible Nitric Oxide Synthase Prevent Damage to Human Lens Epithelial Cells Induced by Interferon-Gamma and Lipopolysaccharide. Biological & Pharmaceutical Bulletin, 29, 2077-2078.
http://dx.doi.org/10.1248/bpb.29.2077
[17]
Linetsky, M., Raghavan, C.T., Johar, K., Fan, X., Monnier, V.M., Vasavada, A.R. and Nagaraj, R.H. (2014) Light-Excited Kynurenines Oxidize Ascorbate and Modify Lens Proteins through the Formation of Advanced Glycation End Products: Implications for Human Lens Aging and Cataract Formation. Journal of Biological Chemistry, 289, 17111-17123.
http://dx.doi.org/10.1074/jbc.M114.554410
[18]
Ornek, K., Karel, F. and Buyukbingol, Z. (2003) May Nitric Oxide Molecule Have a Role in the Pathogenesis of Human Cataract? Experimental Eye Research, 76, 23-27.
http://dx.doi.org/10.1016/S0014-4835(02)00268-3
[19]
Kulaksizoglu, S. and Karalezli, A. (2016) Aqueous Humour and Serum Levels of Nitric Oxide, Malondialdehyde and Total Antioxidant Status in Patients with Type 2 Diabetes with Proliferative Diabetic Retinopathy and Nondiabetic Senile Cataracts. Canadian Journal of Diabetes, 40, 115-119. http://dx.doi.org/10.1016/j.jcjd.2015.07.002
[20]
Li, C.Q. and Wogan, G.N. (2005) Nitric Oxide as a Modulator of Apoptosis. Cancer Letters, 226, 1-15. http://dx.doi.org/10.1016/j.canlet.2004.10.021
[21]
Thiagarajan, R. and Manikandan, R. (2013) Antioxidants and Cataract. Free Radical Research, 47, 337-345. http://dx.doi.org/10.3109/10715762.2013.777155
[22]
Kao, C.L., Chou, C.K., Tsai, D.C., Hsu, W.M., Liu, J.H., Wang, C.S., Lin, J.C., Wu, C.C., Peng, C.H., Chang, C.J. and Chiou, S.H. (2002) Nitric Oxide Levels in the Aqueous Humor in Cataract Patients. Journal of Cataract & Refractive Surgery, 28, 507-512.
[23]
Yousef, M.I., Awad, T.I., Elhag, F.A. and Khaled, F.A. (2007) Study of the Protective Effect of Ascorbic Acid against the Toxicity of Stannous Chloride on Oxidative Damage, Antioxidant Enzymes and Biochemical Parameters in Rabbits. Toxicology, 235, 194-202.
[24]
Matsui, H., Lin, L.R., Ho, Y.S. and Reddy, V.N. (2003) The Effect of up- and down Regulation of MnSOD Enzyme on Oxidative Stress in Human Lens Epithelial Cells. Investigative Ophthalmology & Visual Science, 44, 3467-3475. http://dx.doi.org/10.1167/iovs.02-0830
[25]
Lin, D., Barnett, M., Grauer, L., Robben, J., Jewell. A., Takemoto, L. and Takemoto, D.J. (2005) Expression of Superoxide Dismutase in Whole Lens Prevents Cataract Formation. Molecular Vision, 11, 853-858.
[26]
Olson, B.J. and Markwell, J. (2007) Assays for Determination of Protein Concentration. Current Protocols in Pharmacology. http://dx.doi.org/10.1002/0471141755.pha03as38
[27]
Mushtaq, S., Naqvi, A.Z., Siddiqui, A.A., Carina, P., Shafqat, J. and Ahmed, N. (2007) Changes in Albumin Precursor and Hsp70 Expressions and Their Potential Role in Response to Corneal Epithelial Wound Repair. Proteomics, 7, 463-468.
http://dx.doi.org/10.1002/pmic.200600446
[28]
Oppermann, M., Cols, N., Nyman, T., Helin, J., Saarinen, J., Byman, I., Toran, N., Alaiya, A.A., Bergman, T., Kalkkinen, N., Gonzàlez-Duarte, R. and Jörnvall, H. (2000) Identification of Foetal Brain Proteins by Two-Dimensional Gel Electrophoresis and Mass Spectrometry Comparison of Samples from Individuals with or without Chromosome 21 Trisomy. European Journal of Biochemistry, 267, 4713-4719.
http://dx.doi.org/10.1046/j.1432-1327.2000.01524.x
[29]
Kamaladevi, V., Thekkuttuparambil, L. and Ajith, A. (2016) Association of Grade of Cataract with Duration of Diabetes, Age and Gender in Patients with Type II Diabetes Mellitus. International Journal of Advances in Medicine, 3, 304-308.
[30]
Sayin, N., Kara, N. and Pekel, G. (2015) Ocular Complications of Diabetes Mellitus. World Journal of Diabetes, 6, 92-108.
[31]
Shrestha, R.K. (2011) Ocular Manifestations in Diabetes, a Hospital Based Prospective Study. Nepal Medical College Journal, 13, 254-256. http://dx.doi.org/10.4239/wjd.v6.i1.92
[32]
Astrom, E., Marklund, S.L., Karlsson, K. and Behndig, A. (2003) Involvement of Nitric Oxide in the Formation of In Vitro Diabetic Cataract in Copper-Zinc Superoxide Dismutase Null Mice. Investigative Ophthalmology & Visual Science, 44, 3469.
[33]
Olofsson, E.M., Marklund, S.L. and Behndig, A. (2007) Glucose-Induced Cataract in CuZn-SOD Null Lenses: An Effect of Nitric Oxide? Free Radical Biology and Medicine, 42, 1098-1105. http://dx.doi.org/10.1016/j.freeradbiomed.2007.01.012
[34]
Uçakhan, O.O., Karel, F., Kanpolat, A., Devrim, E. and Durak, I. (2006) Superoxide Dismutase Activity in the Lens Capsule of Patients with Pseudoexfoliation Syndrome and Cataract. Journal of Cataract & Refractive Surgery, 32, 618-622.
http://dx.doi.org/10.1016/j.jcrs.2006.01.018
[35]
Sawada, H., Fukuchi, T. and Abe, H. (2009) Oxidative Stress Markers in Aqueous Humor of Patients with Senile Cataracts. Current Eye Research, 34, 36-41.
http://dx.doi.org/10.1080/02713680802500960
[36]
Giacco, F. and Brownlee, M. (2010) Oxidative Stress and Diabetic Complications. Circulation Research, 107, 1058-1070. http://dx.doi.org/10.1161/CIRCRESAHA.110.223545
