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

摘要/Abstract

摘要：

探讨菜豆多肽（Phaseolus vulgaris Linn peptide，PVP）的抑菌活性及其对脂多糖（LPS）诱导的小鼠单核巨噬细胞RAW264.7中趋化因子和炎症因子转录的抑制作用。应用不同质量浓度的菜豆多肽处理金黄色葡萄球菌和大肠杆菌24 h，检测了最小抑菌浓度（MIC）和抑菌圈直径。使用CCK-8试剂盒检测了菜豆多肽对RAW264.7细胞活力的影响。采用RT-qPCR检测RAW264.7细胞中的趋化因子（趋化因子配体2（CCL2）、巨噬细胞炎症蛋白-2（MIP-2）、趋化因子配体5（CCL5））和炎症因子（诱导型一氧化氮合酶（iNOS）、环氧合酶-2（COX-2）、肿瘤坏死因子-α（TNF-α）、白介素-1β（IL-1β）和IL-6）的转录。采用Western blotting检测NF-κB活化。菜豆多肽对金黄色葡萄球菌和大肠杆菌的MIC均为200 μg/mL。与0 μg/mL组相比，10，50，100，200，300，400，500 μg/mL组的金黄色葡萄球菌和大肠杆菌的抑菌圈直径均呈菜豆多肽剂量依赖性升高（P<0.001）。在1~500 μg/mL的质量浓度范围内，菜豆多肽对RAW264.7细胞活力无明显抑制或促进作用（P>0.05）。与LPS组相比，LPS+10，50，100，200，300，400，500 μg/mL PVP组RAW264.7细胞中CCL2、MIP-2、CCL5、iNOS、COX-2、TNF-α、IL-1β和IL-6 mRNA相对水平以及NF-κB p65相对磷酸化水平均呈菜豆多肽剂量依赖性降低（P<0.05）。本研究表明菜豆多肽具有良好的抑菌活性，可有效抑制LPS诱导的RAW264.7细胞中趋化因子和炎症因子的转录。提示菜豆多肽可能是一种具有良好抑菌和抗炎活性的化妆品添加剂，具有潜在的开发价值。

关键词: 菜豆多肽, 抑菌, 趋化因子, 炎症因子, 脂多糖, 巨噬细胞, RAW264.7细胞

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

中图分类号:

TQ658

辛婷, 李娇娇, 王钊. 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[J]. 日用化学工业（中英文）, 2025, 55(1): 75-83.

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.

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参考文献 21

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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