菜豆多肽的抑菌活性及对趋化因子和炎症因子转录的抑制作用研究

doi:10.3969/j.issn.2097-2806.2025.01.009

Abstract

Abstract:

This study aims to explore the antibacterial activity of Phaseolus vulgaris Linn peptide （PVP） and its inhibitory effect on the transcription of chemokines and inflammatory factors in mouse monocyte macrophages RAW264.7 induced by lipopolysaccharide （LPS）. Different mass concentrations of Phaseolus vulgaris Linn peptide were used to treat Staphylococcus aureus and Escherichia coli for 24 hours, and the minimum inhibitory concentration （MIC） and antibacterial circle diameter were detected. Then the CCK-8 kit was used to detect the effect of Phaseolus vulgaris peptide on the viability of RAW264.7 cells. RT-qPCR was used to detect transcription of chemokines （chemokine ligand 2 （CCL2）, macrophage inflammatory protein-2 （MIP-2）, and chemokine ligand 5 （CCL5）） and inflammatory factors （inducible nitric oxide synthase （iNOS）, cyclooxygenase-2 （COX-2）, tumor necrosis factor-α （TNF-α）, interleukin-1β （IL-1β）, and IL-6）. Western blotting was used to detect NF-κB activation. The MIC of Phaseolus vulgaris Linn peptide against Staphylococcus aureus and Escherichia coli is 200 μg/mL. Compared with the 0 μg/mL group, the antibacterial circle diameters of Staphylococcus aureus and Escherichia coli increase in 10, 50, 100, 200, 300, 400, and 500 μg/mL groups in a dose-dependent manner （P<0.001）. Within the mass concentration range of 1-500 μg/mL, Phaseolus vulgaris Linn peptide has no significant inhibitory or promotional effect on RAW264.7 cell viability （P>0.05）. Compared with the LPS group, CCL2, MIP-2, CCL5, iNOS COX-2, TNF-α, IL-1β and IL-6 mRNA relative levels as well as the relative phosphorylation level of NF-κB p65 in RAW264.7 cells in the LPS+10, 50, 100, 200, 300, 400, 500 μg/mL PVP group are all reduced in a dose-dependent manner （P<0.05）. This study shows that Phaseolus vulgaris Linn peptide has good antibacterial properties activity, which can effectively inhibit the transcription of chemokines and inflammatory factors in RAW264.7 cells induced by LPS. This study suggests that Phaseolus vulgaris Linn peptide may be a cosmetic additive with good antibacterial and anti-inflammatory activities and has potential development value.

Key words: Phaseolus vulgaris Linn peptide, antibacterial, chemokines, inflammatory factors, lipopolysaccharide, macrophages, RAW264.7 cells

CLC Number:

TQ658

Ting Xin, Jiaojiao Li, Zhao Wang. Study on the antibacterial activity of Phaseolus vulgaris Linn peptide and its inhibitory effect on the transcription of chemokines and inflammatory factors[J].China Surfactant Detergent & Cosmetics, 2025, 55(1): 75-83.

Figures/Tables 8

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References 21

[1]	Oseguera-Toledo M E, de Mejia EG, Dia V P, et al. Common bean (Phaseolus vulgaris L.) hydrolysates inhibit inflammation in LPS-induced macrophages through suppression of NF-κB pathways[J]. Food Chem., 2011, 127 (3) : 1175-1185. doi: 10.1016/j.foodchem.2011.01.121 pmid: 25214111
[2]	Ebrahim A E, Abd El-Aziz N K, Elariny E Y T, et al. Antibacterial activity of bioactive compounds extracted from red kidney bean (Phaseolus vulgaris L.) seeds against multidrug-resistant Enterobacterales[J]. Front Microbiol., 2022, 13: 1035586.
[3]	Avagliano C, de Caro C, Cuozzo M, et al. Phaseolus vulgaris extract ameliorates high-fat diet-induced colonic barrier dysfunction and inflammation in mice by regulating peroxisome proliferator-activated receptor expression and butyrate levels[J]. Front Pharmacol., 2022, 13: 930832.
[4]	Korhonen H, Pihlanto A. Food-derived bioactive peptides—opportunities for designing future foods[J]. Curr. Pharm. Des., 2003, 9 (16) : 1297-1308.
[5]	Wong J H, Ng T B. Vulgarinin, a broad-spectrum antifungal peptide from haricot beans (Phaseolus vulgaris)[J]. Int. J. Biochem. Cell Biol., 2005, 37 (8) : 1626-1632.
[6]	Wong J H, Ng T B. Lunatusin, a trypsin-stable antimicrobial peptide from lima beans (Phaseolus lunatus L.)[J]. Peptides, 2005, 26 (11) : 2086-2092.
[7]	López-Barrios L, Antunes-Ricardo M, Gutiérrez-Uribe J A. Changes in antioxidant and antiinflammatory activity of black bean (Phaseolus vulgaris L.) protein isolates due to germination and enzymatic digestion[J]. Food Chem., 2016, 203: 417-424. doi: S0308-8146(16)30209-6 pmid: 26948633
[8]	Gemeinder J L P, Barros N R, Pegorin G S, et al. Gentamicin encapsulated within a biopolymer for the treatment of Staphylococcus aureus and Escherichia coli infected skin ulcers[J]. J. Biomater. Sci. Polym. Ed., 2021, 32 (1) : 93-111.
[9]	Amarowicz R, Dykes G A, Pegg R B. Antibacterial activity of tannin constituents from Phaseolus vulgaris, Fagoypyrum esculentum, Corylus avellana and Juglans nigra[J]. Fitoterapia, 2008, 79 (3) : 217-219. doi: 10.1016/j.fitote.2007.11.019 pmid: 18325686
[10]	Khan M T, Usman M, Nawaz H A, et al. Formulation of Phaseolus vulgaris L. cream and its characterization[J]. Pak. J. Pharm. Sci., 2020, 33 (2) : 815-820.
[11]	Fonseca-Hernández D, Lugo-Cervantes E D C, Escobedo-Reyes A, et al. Black bean (Phaseolus vulgaris L.) polyphenolic extract exerts antioxidant and antiaging potential[J]. Molecules, 2021, 26 (21) : 6716.
[12]	Chen Y, Liu S, Wu L, et al. Epigenetic regulation of chemokine (CC-motif) ligand 2 in inflammatory diseases [J]. Cell Prolif., 2023, 56 (7) : e13428.
[13]	Son D, Na Y, Cho W S, et al. Differentiation of skin sensitizers from irritant chemicals by interleukin-1α and macrophage inflammatory protein-2 in murine keratinocytes[J]. Toxicol. Lett., 2013, 216 (1) : 65-71. doi: 10.1016/j.toxlet.2012.10.017 pmid: 23178550
[14]	Kim D, Kim J, Kwon S, et al. Regulation of macrophage inflammatory protein-2 gene expression in response to 2, 4-dinitrofluorobenzene in RAW 264.7 cells[J]. BMB Rep., 2008, 41 (4) : 316-321.
[15]	Lin J, Xu Y, Guo P, et al. CCL5/CCR5-mediated peripheral inflammation exacerbates blood-brain barrier disruption after intracerebral hemorrhage in mice[J]. J. Transl. Med., 2023, 21 (1) : 196.
[16]	Kandhaya-Pillai R, Yang X, Tchkonia T, et al. TNF-α/IFN-γ synergy amplifies senescence-associated inflammation and SARS-CoV-2 receptor expression via hyper-activated JAK/STAT1[J]. Aging Cell, 2022, 21 (6) : e13646.
[17]	Perez-Hernandez L M, Nugraheni K, Benohoud M, et al. Starch digestion enhances bioaccessibility of anti-inflammatory polyphenols from borlotti beans (Phaseolus vulgaris)[J]. Nutrients, 2020, 12 (2) : 295.
[18]	Hayden M S, Ghosh S. Shared principles in NF-κB signaling[J]. Cell, 2008, 132 (3) : 344-362.
[19]	Lawrence T. The nuclear factor NF-κB pathway in inflammation[J]. Cold. Spring Harb. Perspect. Biol., 2009, 1 (6) : a001651.
[20]	Shen Y, Boulton A P R, Yellon R L, et al. Skin manifestations of inborn errors of NF-κB[J]. Front Pediatr., 2022, 10: 1098426.
[21]	Wang Y, Wang L, Wen X, et al. NF-κB signaling in skin aging[J]. Mech. Ageing Dev., 2019, 184: 111160.

Gene	Primer sequence
CCL2	F: 5'-AAGCCAGCTCTCTCTTCCTC-3'
CCL2	R: 5'-CCTCTCTCTTTGAGCTTGGTG-3'
MIP-2	F: 5'-TGTCAATGCCTGAAGACCCTGCC-3'
MIP-2	R: 5'-AACTTTTTGACCGCCCTTGAGAGTGG-3'
CCL5	F: 5'-ATCTTGCAGTCGTGTTTGTCAC-3'
CCL5	R: 5'-GAAATGCTGATTTCTTGGGTTT-3'
iNOS	F: 5'-GATCAATAACCTGAAGCCCG-3'
iNOS	R: 5'-GCCCTTTTTTGCTCCATAGG-3'
COX-2	F: 5'-TCCAATCGCTGTACAAGCAG-3'
COX-2	R: 5'-TCCCCAAAGATAGCATCTGG-3'
TNF-α	F: 5'-CGTCCATTCTTGCCCAAAC-3'
TNF-α	R: 5'-AGCCCTGAGCCCTTAATTC-3'
IL-1β	F: 5'-GCTGAAAGCTCTCCACCTCAATG-3'
IL-1β	R: 5'-TGTCGTTGCTTGGTTCTCCTTG-3'
IL-6	F: 5'-CCAAGGATGATGCCACTT-3'
IL-6	R: 5'-CATCTCCTTTCTCAGTGC-3'
β-actin	F: 5'-GTGCCGCCTGGAGAAACCT-3'
β-actin	R: 5'-AAGTCGCAGGAGACAACC-3'

Group	Antibacterial circle diameter/mm		Group	Antibacterial circle diameter/mm
Group	Staphylococcus aureus	Escherichia coli	Group	Staphylococcus aureus	Escherichia coli
0 μg/mL PVP	0.00±0.00	0.00±0.00	300 μg/mL PVP	17.19±1.82 ^***	17.55±1.79 ^***
1 μg/mL PVP	0.00±0.00	0.00±0.00	400 μg/mL PVP	18.80±1.91 ^***	21.27±1.45 ^***
10 μg/mL PVP	5.11±0.07 ^***	6.04±0.13 ^***	500 μg/mL PVP	22.96±1.81 ^***	22.84±1.51^***
50 μg/mL PVP	7.36±0.54 ^***	8.97±1.17 ^***	F	306.448	455.951
100 μg/mL PVP	9.77±0.63 ^***	10.39±0.68 ^***	P	<0.001	<0.001
200 μg/mL PVP	14.73±1.04 ^***	14.34±1.00 ^***

Group	mRNA相对水平
Group	iNOS	COX-2	TNF-α	IL-1β	IL-6
Control	1.00±0.06	1.00±0.05	1.00±0.02	1.00±0.03	1.00±0.02
LPS	10.32±0.32 ^*	15.78±0.94 ^*	20.86±0.96 ^*	18.70±0.85 ^*	14.29±1.32 ^*
LPS+1 μg/mL PVP	10.19±0.54 ^*	16.46±1.10 ^*	20.17±1.21 ^*	17.99±1.12 ^*	14.86±0.96 ^*
LPS+10 μg/mL PVP	8.59±0.53 ^*#	13.39±1.25 ^*#	16.79±0.87 ^*#	14.33±1.07 ^*#	11.97±1.00 ^*#
LPS+50 μg/mL PVP	5.98±0.46 ^*#	11.29±0.77 ^*#	11.79±0.42 ^*#	10.15±0.91 ^*#	7.82±0.67 ^*#
LPS+100 μg/mL PVP	3.96±0.10 ^*#	7.54±0.52 ^*#	8.13±0.38 ^*#	6.00±0.31 ^*#	4.85±0.23 ^*#
LPS+200 μg/mL PVP	3.46±0.12 ^*#	4.52±0.17 ^*#	4.79±0.15 ^*#	4.07±0.18 ^*#	3.70±0.13 ^*#
LPS+300 μg/mL PVP	2.56±0.19 ^*#	3.25±0.02 ^*#	3.46±0.07 ^*#	2.73±0.09 ^*#	2.76±0.09 ^*#
LPS+400 μg/mL PVP	2.26±0.09 ^*#	2.40±0.22 ^*#	2.48±0.12 ^*#	2.01±0.09 ^*#	2.16±0.15 ^*#
LPS+500 μg/mL PVP	1.72±0.16 ^*#	1.52±0.15 ^*#	1.68±0.10 ^*#	1.38±0.06 ^*#	1.47±0.16 ^*#
F	765.608	472.780	1 026.024	708.349	409.810
P	<0.001	<0.001	<0.001	<0.001	<0.001

Study on the antibacterial activity of Phaseolus vulgaris Linn peptide and its inhibitory effect on the transcription of chemokines and inflammatory factors

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