Abstract
Obesity is one of the major contributing factors responsible for the onset of various metabolic syndrome. In obesity-related metabolic syndrome, increased oxidative stress in accumulated fat is a key pathogenic mechanism. The aim of this study is to investigate the lipid-lowering and antioxidant effects of RF13 peptide derived from vacuolar protein sorting associated protein 26B (VPS26B) in high fat diet (HFD) induced zebrafish larvae. Based on the arrangement, composition, and bioinformatics analysis of the VPS26B protein sequence, we predicted and synthesized a short peptide, 1RRGKGGRRVTMSF13 (RF13). In vitro analysis (DPPH, ABTS and nitric oxide assay) of RF13 on showed its potential antioxidant properties and radical scavenging capacity. Rat skeletal muscle cells (L6) and human erythrocyte cells are used for the evaluation of RF13 cytotoxicity at differing concentrations showed no cytotoxic effect. Zebrafish larvae feeding HFD, on day 4 post fertilization (dpf) to 6 dpf, there was an increase in production of reactive oxygen species (ROS) and lipid accumulation, which showed decreased expression of antioxidative enzymes such as glutathione S-transferase (GST), glutathione reductase (GR) and glutathione peroxidase (GPx). However, RF13 treatment has significantly reduced the total cholesterol and triglycerides levels, enhanced antioxidant enzyme levels, and decreased lipid peroxidation. In addition, RF13 treatment has significantly decreased lipid accumulation, intracellular ROS and apoptosis in zebrafish larvae as compared to the HFD group. Furthermore, RF13 significantly downregulated the lipid metabolizing genes such as CCAAT/enhancer-binding protein-α (C/EBP-α), sterol regulatory element-binding protein 1 (SREBP1) and fatty acid synthase (FAS), while they upregulated the antioxidant-related genes. These results suggests that RF13 peptides can be used as functional foods and natural drugs to enhance the antioxidant ability and also to alleviate lipid accumulation by regulating lipid metabolism.
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Data Availability Statement
Data will be made available on reasonable request.
Abbreviations
- VPS26B:
-
Vacuolar protein sorting associated protein 26B
- HFD:
-
High fat diet
- FAS:
-
Fatty acid synthase
- LPL:
-
Lipoprotein lipase
- PPAR :
-
Peroxisome proliferator-activated receptor
- C/EBP-α:
-
CCAAT/enhancer-binding protein-α
- SREBP1:
-
Sterol regulatory element-binding protein 1
- ROS:
-
Reactive oxygen species
- GR:
-
Glutathione reductase
- GPx:
-
Glutathione peroxidase
- SOD:
-
Superoxide dismutase
- CAT:
-
Catalase
- DPPH:
-
2,2-diphenyl-1-picrylhydrazyl
- ABTS:
-
2,2 -azino-bis(3-ethylbenzothiazoline-6-sulfonic acid
- HFD:
-
High fat diet
- DCFDA:
-
2’-7’-dichlorofluorescein diacetate
- DPPP:
-
Diphenyl-1-pyrenylphosphine
- RNS:
-
Reactive nitrogen species
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The authors acknowledge Dr. Kanchana K. Mala for providing necessary help in performing haemolytic assay and associated ethical clearance (885/IEC/2015).
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Conceptualization, Methodology, Formal analysis and investigation Writing-original draft preparation: AJ, MV; Conceptualization, Formal analysis, Funding acquisition, Supervision: JA.
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This research does not involve any human or animal objects; however, we have collected human blood from healthy volunteers for the hemolytic assay performed in this study. The blood from the healthy individuals was collected as per the guidelines of the Institutional Ethical Committee regulation (Ethical Clearance No. 885/IEC/2015). For blood collection, informed consent was obtained from all the participated adults in written form. All authors are aware of the details of their research work that are presented in the current manuscript and gave their consent to publication.
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Guru, A., Velayutham, M. & Arockiaraj, J. Lipid-Lowering and Antioxidant Activity of RF13 Peptide From Vacuolar Protein Sorting-Associated Protein 26B (VPS26B) by Modulating Lipid Metabolism and Oxidative Stress in HFD Induced Obesity in Zebrafish Larvae. Int J Pept Res Ther 28, 74 (2022). https://doi.org/10.1007/s10989-022-10376-3
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DOI: https://doi.org/10.1007/s10989-022-10376-3