Proteomics and Molecular Docking Analyses Reveal the Bio-Chemical and Molecular Mechanism Underlying the Hypolipidemic Activity of Nano-Liposomal Bioactive Peptides in 3T3-L1 Adipocytes

Sucheewin Krobthong; Yodying Yingchutrakul; Patompon Wongtrakoongate; Hathaichanok Chuntakaruk; Thanyada Rungrotmongkol; Chartchai Chaichana; Thanisorn Mahatnirunkul; Thitikorn Chomtong; Kiattawee Choowongkomon; Chanat Aonbangkhen

doi:10.3390/foods12040780

Proteomics and Molecular Docking Analyses Reveal the Bio-Chemical and Molecular Mechanism Underlying the Hypolipidemic Activity of Nano-Liposomal Bioactive Peptides in 3T3-L1 Adipocytes

Foods. 2023 Feb 10;12(4):780. doi: 10.3390/foods12040780.

Authors

Affiliations

¹ Center of Excellence in Natural Products Chemistry (CENP), Department of Chemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.
² Center for Neuroscience, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
³ National Omics Center, NSTDA, Pathum Thani 12120, Thailand.
⁴ Department of Biochemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
⁵ Center of Excellence in Biocatalyst and Sustainable Biotechnology, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Bangkok 10330, Thailand.
⁶ Program in Bioinformatics and Computational Biology, Graduate School, Chulalongkorn University, Bangkok 10330, Thailand.
⁷ Siriraj Center of Research Excellence for Diabetes and Obesity (SiCORE-DO), Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.
⁸ National Nanotechnology Center, NSTDA, Pathum Thani 12120, Thailand.
⁹ Department of Biochemistry, Faculty of Science, Kasetsart University, Bangkok 10900, Thailand.

Abstract

Obesity is a global health concern. Physical activities and eating nutrient-rich functional foods can prevent obesity. In this study, nano-liposomal encapsulated bioactive peptides (BPs) were developed to reduce cellular lipids. The peptide sequence NH₂-PCGVPMLTVAEQAQ-CO₂H was chemically synthesized. The limited membrane permeability of the BPs was improved by encapsulating the BPs with a nano-liposomal carrier, which was produced by thin-layer formation. The nano-liposomal BPs had a diameter of ~157 nm and were monodispersed in solution. The encapsulation capacity was 61.2 ± 3.2%. The nano-liposomal BPs had no significant cytotoxicity on the tested cells, keratinocytes, fibroblasts, and adipocytes. The in vitro hypolipidemic activity significantly promoted the breakdown of triglycerides (TGs). Lipid droplet staining was correlated with TG content. Proteomics analysis identified 2418 differentially expressed proteins. The nano-liposomal BPs affected various biochemical pathways beyond lipolysis. The nano-liposomal BP treatment decreased the fatty acid synthase expression by 17.41 ± 1.17%. HDOCK revealed that the BPs inhibited fatty acid synthase (FAS) at the thioesterase domain. The HDOCK score of the BPs was lower than that of orlistat, a known obesity drug, indicating stronger binding. Proteomics and molecular docking analyses confirmed that the nano-liposomal BPs were suitable for use in functional foods to prevent obesity.

Keywords: HDOCK; adipocyte; fatty acid synthase; glycerol; lipolysis; liposome; nanoparticles; orlistat.

Grants and funding

This research project was supported by Second Century Fund (C2F), Chulalongkorn University. This project was financially supported by grants from the Innovation Policy Council by Program Management Unit for Human Resources and Institutional Development, Research, and Innovation (PMU-B; grant number B05F640047). C.A. would like to thank the Research Grant for New Scholar (grant no. RGNS 64–004) by the Office of the Permanent Secretary, Ministry of Higher Education, Science, Research, and Innovation, the Development of Chula New Faculty Staff (grant no. DNS_66_003_23_001_2) as well as the Fundamental Fund (FF66, grant no. BCG66230008) through the Chulalongkorn University Ratchadaphiseksomphot Endowment Fund, the Asahi Glass Foundation (to C.A.), and the 29th Science and Technology Research Grant 2022 from the Thailand Toray Science Foundation (to C.A.). K.C. wishes to thank the Kasetsart University Research and Development Institute (grant no. FF (KU)25.64). C.C. wishes to thank the grant by the Agricultural Research Development Agency (grant no. PRP6405030610).