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Proteomic Analysis of Plasma Proteins in Diabetic Rats by 2D Electrophoresis and MALDI-TOF-MS

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Abstract

Despite tremendous advances in our understanding of the molecular basis of diabetes mellitus, substantial gaps still remain in our understanding of disease pathogenesis and in the development of effective strategies for early diagnosis and treatment. The proteomic approach has offered many opportunities and challenges in identifying new marker proteins and therapeutic targets, i.e., using 2D-polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionisation-time of flight mass spectrometry. The differential protein expressions were analyzed in alloxan-induced diabetic rats treated with Cynodon dactylon leaf extract. The plant extract was administered for 15 days that resulted in a significant increase in plasma insulin and C-peptide levels. We have also identified four differentially expressed proteins from rat plasma. These four diabetes-associated proteins were broadly classified into three groups as per their function: (1) lipid metabolism-associated protein (Apo A-IV), (2) antioxidant activity-related proteins [preprohaptoglobin and heat shock proteins B8 (HspB8)], and (3) muscle function-related protein (TPM3). Apo A-IV, HspB8, and preprohaptoglobin may play a key role in the recovery of diabetes mellitus and also prevent the diabetes-associated complications such as prevention of oxidative stress due to free radical and free hemoglobin. These results show the value of proteomic approach in identifying the potential markers that may eventually serve as diagnostic markers or therapeutic targets.

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Acknowledgment

The first author (D.K.) deeply owes his sincere thanks to Merck Millipore-MSPL, Bengaluru, KA, India for granting special permission to PhD. The authors thankful to the Department of Biotechnology, PRIST University, Thanjavur, Tamil Nadu, India for providing lab facilities. The authors are grateful to Prof. Dipankar Chatterji and Mr. Sathish K, Molecular Biophysics Unit, Indian Institute of Science (IISc), Bangalore, Karnataka, India for providing MALDI-TOF-MS/MS facility, and for their invaluable and helpful discussions.

Conflict of interest

The authors are not having any conflict of interest related to this paper.

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Authors and Affiliations

  1. Department of Biotechnology, PRIST University, Thanjavur, Tamil Nadu, 613 403, India

    D. Karthik, S. Ilavenil, B. Kaleeswaran & S. Ravikumar

  2. Merck Millipore, Merck Specialities Private Limited (MSPL), Bengaluru, Karnataka, 560 058, India

    D. Karthik & S. Sunil

  3. Faculty of Biotechnology, PRIST University, Thanjavur, 613 403, Tamilnadu, India

    S. Ravikumar

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  1. D. Karthik
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Correspondence to S. Ravikumar.

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Karthik, D., Ilavenil, S., Kaleeswaran, B. et al. Proteomic Analysis of Plasma Proteins in Diabetic Rats by 2D Electrophoresis and MALDI-TOF-MS. Appl Biochem Biotechnol 166, 1507–1519 (2012). https://doi.org/10.1007/s12010-012-9544-8

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