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. 2024 Sep 27:24:101863.
doi: 10.1016/j.fochx.2024.101863. eCollection 2024 Dec 30.

Immunomodulatory peptides from sturgeon cartilage: Isolation, identification, molecular docking and effects on RAW264.7 cells

Shuchan Li  1 , Miaoqing An  1 , Yuxuan Zhao  1 , Wenjun Zhao  1 , Pan Li  1 , Bing Du  1
Affiliations

Immunomodulatory peptides from sturgeon cartilage: Isolation, identification, molecular docking and effects on RAW264.7 cells

Shuchan Li et al. Food Chem X. .

Abstract

Sturgeons (Acipenseridae), ancient fish known for their caviar, produce underutilized by-products like protein-rich cartilage, which is a source of high-quality bioactive peptides. This study investigates immunomodulatory peptides from sturgeon cartilage hydrolysates mechanisms. The study found that sturgeon cartilage hydrolysate F2-7 and its key peptides(DHVPLPLP and HVPLPLP)significantly promoted RAW267.4 cell proliferation, NO release, and phagocytosis (P < 0.001).Additionally, western blotting confirmed that F2-7 enhances immune response by increasing the expression of P-IKKα/β, IΚΚ, p65, and P-p65 proteins in the NF-κB signalling pathway (P < 0.01). Molecular docking further demonstrated that DHVPLPLP and HVPLPLP bind to NF-κB pathway proteins via hydrogen bonding, with low estimated binding energies (-2.75 and -1.64; -6.04 and -4.75 kcal/mol), thus establishing their role as key immune peptides in F2-7. Therefore, DHVPLPLP and HVPLPLP have the potential to be developed as dietary supplements for immune enhancement. Their ability to enhance immune function provides a theoretical basis for novel immune supplements.

Keywords: Immune regulation; Molecular docking; Mouse monocyte macrophage leukaemia cells (RAW264.7); Peptides; Sturgeon cartilage.

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Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Unlabelled Image
Graphical abstract
Fig. 1
Fig. 1
(A) Sephadex G-15 filtration, (B) Ion-exchange chromatography, (C) RT-HPLC purification of fraction F2, (D) Effect of F2 on RAW264.7 cell viability, (E) NO production, and (F) neutral red phagocytosis. DMEM and LPS-treated groups served as negative and positive controls, respectively.Data are presented as the mean ± SD (n = 5). ⁎P < 0.05 vs. NC, ⁎⁎P < 0.01 vs. NC, ⁎⁎⁎P < 0.001 vs. NC; #P < 0.05 vs. LPS, ##P < 0.01 vs. LPS, ###P < 0.001 vs. LPS. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Fig. 2
Fig. 2
Effects of RT-HPLC purified fractions on the (A) viability of RAW 264.7 cells, production of (B) NO, and (C) phagocytosis of neutral red, (D) TNF-α, (E) IL-1β, and (F) IL-6 production. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Fig. 3
Fig. 3
Effects of F2–7 fraction on RAW 264.7 cells morphology (A), (B) NO secretion, (C) neutral red phagocytosis, (D) TNF-α, (E) IL-1β, and (F) IL-6 production. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Fig. 4
Fig. 4
(A) (B) (C) (D) RAW264.7 cell-related protein expression. (E) NF-κB-related protein blotting map.
Fig. 5
Fig. 5
The distribution of peptide length and quantity of Sturgeon cartilage peptides F2–7 in external layer pie plot, as well as the distribution of its bioactive peptides (score > 0.5) at internal layer pie plot.
Fig. 6
Fig. 6
The interaction of peptides identified from F2–7 with IKK by molecular docking. A, C are 3D diagrams of specific residues of the peptides interacting with IKK, and the orange dashed line indicates the hydrogen bonding.B, D show the active pockets formed between the peptide and the receptor proteins. (A, B) Molecular docking results of DHVPLPLP with IKK. (C, D) Molecular docking results of HVPLPLP with IKK.
Fig. 7
Fig. 7
The interaction of peptides identified fromF2–7 with p65 by molecular docking. (A, B) Molecular docking result of DHVPLPLP with p65. (C, D) Molecular docking result of HVPLPLP with p65.
Fig. 8
Fig. 8
(A) Effect of synthetic peptides on RAW264.7 cell viability, (B) NO secretion, (C) neutral red phagocytosis. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
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