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Royal Jelly Proteins Inhibit Macrophage Proliferation: Interactions with Native- and Oxidized-Low Density Lipoprotein

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Abstract

Macrophage proliferation is known to correlate with macrophage accumulation in atherosclerotic plaque, and therefore its inhibition and secondary reduction of plaque inflammation may have therapeutic beneficial effects on atherosclerosis. Recently, we reported that a peptide corresponding to positions 41–51 of royalisin (which consists of 51 amino acid residues), a potent antibacterial protein contained in royal jelly (RJ), can specifically bind to oxidized LDL (Ox-LDL), a major components of atherosclerotic lesions. Here, we investigated the interaction of RJ proteins including royalisin with LDL and Ox-LDL. Measurement of LDL oxidation by the production of thiobarbituric acid reactive substances and conjugated dienes, and by electrophoretic mobility on polyacrylamide gel electrophoresis showed that RJ proteins including royalisin and the degradation products of major RJ protein (MRJP) 1 and MRJP3 can induce oxidation of LDL and Ox-LDL. Surface plasmon resonance experiments showed that these RJ proteins can exhibit much higher binding affinity to LDL than Ox-LDL (the equilibrium dissociation constant, KD = 8.35 and 49.65 μg proteins/mL for LDL and Ox-LDL, respectively). Experiments using cultured mouse J774A.1 macrophage cells proved that these RJ proteins can inhibit macrophage proliferation markedly and concentration-dependently, regardless of the absence or presence of LDL and Ox-LDL, but hardly affect lipid accumulation in macrophages. These results suggest that RJ proteins including royalisin and degradation products of MRJP1/MRJP3 may have therapeutic beneficial effects on atherosclerosis owing to the reduction of plaque inflammation. Further studies of these RJ proteins may lead to the discovery of novel anti-atherosclerotic drugs.

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Acknowledgements

This research did not receive any specific grant from funding agencies in the public, commercial, or not-profit sectors. The authors thank Ms. Hiromi Yoshida (Common Instrument Center, Institute of Development, Aging and Cancer, Tohoku University) for surface plasmon resonance measurement. We would like to thank Editage (Tokyo, Japan) for editing and reviewing this manuscript for English language.

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Authors and Affiliations

  1. Faculty of Pharmacy, Iryo Sosei University, 5-5-1, Chuodai-Iino, Iwaki, Fukushima, 970-8551, Japan

    Akira Sato, Hiroto Unuma & Keiichi Ebina

  2. Graduate School of Life Science and Engineering, Iryo Sosei University, 5-5-1, Chuodai-Iino, Iwaki, Fukushima, 970-8551, Japan

    Akira Sato & Keiichi Ebina

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  1. Akira Sato
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AS and KE conceived and designed the experiments. AS and HU carried out the experiments and analyzed the data. AS wrote the manuscript.

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Correspondence to Akira Sato.

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Sato, A., Unuma, H. & Ebina, K. Royal Jelly Proteins Inhibit Macrophage Proliferation: Interactions with Native- and Oxidized-Low Density Lipoprotein. Protein J 40, 699–708 (2021). https://doi.org/10.1007/s10930-021-09998-1

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