蒲公英多酚的抑菌和抗炎活性研究

doi:10.3969/j.issn.2097-2806.2025.05.012

摘要/Abstract

摘要：

为了揭示蒲公英多酚（DP）对常见皮肤病原菌的抑菌活性及其抗炎活性。使用不同质量浓度的蒲公英多酚处理6种常见皮肤病原菌24 h，测量最小抑菌浓度（MIC）和抑菌圈直径。使用CCK-8试剂盒检测蒲公英多酚对RAW264.7细胞的毒性。将RAW264.7细胞分为Blank组、LPS组、LPS+10 DP组、LPS+50 DP组、LPS+100 DP组、LPS+200 DP组、LPS+400 DP组、LPS+800 DP组和LPS+1 600 DP组。根据分组使用生理盐水、LPS（1 μg/mL）和不同质量浓度的DP处理细胞24 h。通过可见分光光度法测量NO含量。通过RT-qPCR检测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（IL）-1β和IL-6的mRNA水平。通过Western blotting检测核因子-κB（NF-κB） p65和p38丝裂原活化蛋白激酶（MAPK）磷酸化水平。免疫荧光染色观察NF-κB p65的细胞核转位情况。结果显示，DP对金黄色葡萄球菌、表皮葡萄球菌、大肠埃希菌、铜绿假单胞菌、普通变形杆菌和痤疮丙酸杆菌的的MIC分别为400，400，400，800，800和400 μg/mL。当DP质量浓度≥800 μg/mL时，6种病原菌均有明显的抑菌圈（P<0.001）。DP质量浓度在10~1 600 μg/mL范围内对RAW264.7细胞无毒（P>0.05）。与LPS组相比，LPS+100 DP组、LPS+200 DP组、LPS+400 DP组、LPS+800 DP组和LPS+1 600 DP组RAW264.7细胞中NO含量及iNOS、COX-2、TNF-α、IL-1β和IL-6 mRNA相对水平均降低（P<0.05），NF-κB p65和p38 MAPK相对磷酸化水平降低（P<0.05），并且NF-κB p65细胞核转位减少。上述结果说明蒲公英多酚对常见皮肤病原菌具有抑菌活性，并且可抑制LPS诱导的RAW264.7细胞炎症及NF-κB p65和p38 MAPK信号通路的活化。

关键词: 蒲公英多酚, 皮肤常见病原菌, 抑菌, 抗炎, 核因子-κB, p38丝裂原活化蛋白激酶

Abstract:

The study aimed to reveal the antibacterial activity of dandelion polyphenols （DP） against common skin pathogens and anti-inflammatory activity. In this study, six common skin pathogens were treated with different concentrations of dandelion polyphenols for 24 hours, and the minimum inhibitory concentration （MIC） and the diameter of bacteriostatic zone were measured. CCK-8 kit was used to detect the toxicity of dandelion polyphenols to RAW264.7 cells. RAW264.7 cells were divided into Blank group, LPS group, LPS+10 DP group, LPS+50 DP group, LPS+100 DP group, LPS+200 DP group, LPS+400 DP group, LPS+800 DP group and LPS+1 600 DP group. The cells were treated with saline, LPS （1 μg/mL） and different concentrations of dandelion polyphenols for 24 hours. The content of NO was measured by visible spectrophotometry. The mRNA levels of inducible nitric oxide synthase （iNOS）, cyclooxygenase-2 （COX-2）, tumor necrosis factor-α （TNF-α）, interleukin （IL）-1β and IL-6 were detected by RT-qPCR. The phosphorylation levels of NF-kappa B （NF-κB） p65 and p38 mitogen activated protein kinase （MAPK） were detected by Western blotting. The nuclear translocation of NF-κB p65 was observed by immunofluorescence staining. The results show that the MIC of dandelion polyphenols against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Pseudomonas aeruginosa, Proteus vulgaris and Cutibacterium acnes are 400, 400, 400, 800, 800 and 400 μg/mL, respectively. When the concentration of dandelion polyphenols is over 800 μg/mL, all the six pathogens have obvious bacteriostatic circles （P<0.001）. The concentration of dandelion polyphenols in the range of 10 to 1 600 μg/mL is non-toxic to RAW264.7 cells （P>0.05）. Compared with LPS group, NO content and relative levels of iNOS, COX-2, TNF-α, IL-1β and IL-6 mRNA in RAW264.7 cells in LPS+100 DP group, LPS+200 DP group, LPS+400 DP group, LPS+800 DP group and LPS+1 600 DP group decrease （P<0.05）, the relative phosphorylation levels of NF-κB p65 and p38 MAPK decrease （P<0.05）, and the nuclear translocation of NF-κB p65 decreases. The above results indicate that dandelion polyphenols have antibacterial activity against common skin pathogens, and can inhibit LPS-induced RAW264.7 cell inflammation and the activation of NF-κB p65 and p38 MAPK signaling pathways.

Key words: dandelion polyphenols, common skin pathogens, bacteriostasis, anti-inflammation, nuclear factor-κB, p38 mitogen-activated protein kinase

中图分类号:

TQ658

谢诗雅, 刘明巍, 肖雷, 胡绍霖. 蒲公英多酚的抑菌和抗炎活性研究[J]. 日用化学工业（中英文）, 2025, 55(5): 633-639.

Shiya Xie, Mingwei Liu, Lei Xiao, Shaolin Hu. Study on the antibacterial activity and anti-inflammatory activity of dandelion polyphenols[J]. China Surfactant Detergent & Cosmetics, 2025, 55(5): 633-639.

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Gene	Primer sequence
iNOS	F: 5′-AGCACTTTGGGTGACCACCAGGA-3′
iNOS	R: 5′-AGCTAAGTATTAGAGCGGCGGCA-3′
COX-2	F: 5′-AGAAGGAAATGGCTGCAGAA-3′
COX-2	R: 5′-GCTCGGCTTCCAGTATTGAG-3′
TNF-α	F: 5′-CTTCAGCCCCAGCAGTGTATTCTTT-3′
TNF-α	R: 5′-GAGAACCTGGGAGTAGACAAGA-3′
IL-1β	F: 5′-ACCTTCCAGGATGAGGACATGA-3′
IL-1β	R: 5′-CTAATGGGAACGTCACACACCA-3′
IL-6	F: 5′-CACATGTTCTCTGGGAAATCG-3′
IL-6	R: 5′-TTGTATCTCTGGAAGTTTCAGATTGTT-3′
β-actin	F: 5′-TGGAATCCTGTGGCATCCATGAAAC-3′
β-actin	R: 5′-TAAAACGCAGCTCAGTAACAGTCCG-3′