积雪草活性成分槲皮素通过介导STAT3磷酸化抑制IL-23/IL-17A炎症轴发挥抗银屑病作用

doi:10.12122/j.issn.1673-4254.2025.01.12

南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (1): 90-99.doi: 10.12122/j.issn.1673-4254.2025.01.12

积雪草活性成分槲皮素通过介导STAT3磷酸化抑制IL-23/IL-17A炎症轴发挥抗银屑病作用

刘青¹(), 刘敬¹, 郑逸航¹, 雷金², 黄建华¹, 刘思妤¹, 刘芳¹, 彭群龙¹, 张远芳¹, 王俊杰¹, 李玉娟²()

^1.湘南学院，药学院，湖南郴州 423000
^2.湘南学院，基础医学院，湖南郴州 423000

收稿日期:2024-06-27 出版日期:2025-01-20 发布日期:2025-01-20
通讯作者: 李玉娟 E-mail:liuqing@xnu.edu.cn;Li_yujuan1001@163.com
作者简介:刘青，副教授，E-mail: liuqing@xnu.edu.cn
基金资助:
湖南省教育厅科学研究优秀青年项目(23B0771);湖南省自然科学基金(2024JJ7522);湖南省卫生健康委科研计划课题(D202302048964);湖南省双一流应用特色学科药学学科(湘教通〔2022〕351号);生物医药产业化中试技术研发公共服务平台(湘南学院科发〔2022〕3号);郴州国家可持续发展议程创新示范区建设省级专项，南岭优势及特色药用植物全产业链关键技术研发(2023sfq04)

Quercetin mediates the therapeutic effect of Centella asiatica on psoriasis by regulating STAT3 phosphorylation to inhibit the IL-23/IL-17A axis

Qing LIU¹(), Jing LIU¹, Yihang ZHENG¹, Jin LEI², Jianhua HUANG¹, Siyu LIU¹, Fang LIU¹, Qunlong PENG¹, Yuanfang ZHANG¹, Junjie WANG¹, Yujuan LI²()

^1.College of Pharmacy, Xiangnan University, Chenzhou 423000, China
^2.College of Basic Medical Sciences, Xiangnan University, Chenzhou 423000, China

Received:2024-06-27 Online:2025-01-20 Published:2025-01-20
Contact: Yujuan LI E-mail:liuqing@xnu.edu.cn;Li_yujuan1001@163.com

摘要/Abstract

摘要：

目的探讨积雪草抗银屑病活性成分及其作用机制。方法利用TCMSP、PharmMapper等数据库获取积雪草化合物和靶点，利用GeneCards数据库筛选银屑病相关的潜在靶点，Cytoscape 3.10.0软件导入积雪草活性成分、银屑病共同靶点制作“药物-活性成分-作用靶点”网络图，使用STRING数据库构建PPI网络，借助DAVID数据库进行通路富集分析。通过网络药理学筛选得到积雪草主要活性成分槲皮素、积雪草苷、积雪草酸，并将这3种活性成分（7.5、15、30、60 μmol/L）作用于脂多糖（LPS）刺激RAW264.7小鼠巨噬细胞炎症模型，Griess法检测细胞NO水平评价抗炎活性。ELISA检测TNF-α、IL-6细胞促炎因子的表达，RT-qPCR测定细胞IL-23、IL-17A、TNF-α、IL-6 mRNA的表达，Western blotting检测细胞p-STAT3（Tyr705）、p-STAT3（Ser727）的表达变化。结果网络药理学研究分析获取积雪草靶点139个，银屑病靶点4604个，PPI网络图显示CASP3、EGFR、PTGS2和ESR1为治疗银屑病的关键靶点，分析得到槲皮素、积雪草苷、积雪草酸可能是积雪草抗银屑病关键活性成分。KEGG结果显示与癌症、IL-17以及MAPK信号通路相关。槲皮素、积雪草苷、积雪草酸处理细胞炎症模型，发现槲皮素表现出明显的抗炎活性，细胞NO水平降低（P<0.05）。与LPS组相比，槲皮素处理后细胞中TNF-α，IL-6促炎因子分泌减少（P<0.05），IL-23、IL-17A、TNF-α、IL-6 mRNA表达水平降低（P<0.05），STAT3磷酸化的两个位点（Tyr705、Ser727）蛋白表达均明显下调（P<0.05）。结论积雪草抗银屑病主要活性成分为槲皮素、积雪草苷和积雪草酸，其中关键成分槲皮素通过抑制NO产生、炎症因子TNF-α、IL-6的表达，并通过介导STAT3磷酸化调控IL-23/IL-17A炎症轴抑制炎症反应发挥抗银屑病作用。

关键词: 积雪草, 银屑病, 槲皮素, IL-23/IL-17A, 网络药理学

Abstract:

Objective To explore the active components that mediate the therapeutic effect of Centella asiatica on psoriasis and their therapeutic mechanisms. Methods TCMSP, TCMIP, PharmMapper, Swiss Target Prediction, GeneCards, OMIM and TTD databases were searched for the compounds in Centella asiatica and their targets and the disease targets of psoriasis. A drug-active component-target network and the protein-protein interaction network were constructed, and DAVID database was used for pathway enrichment analysis. In a RAW264.7 macrophage model of LPS-induced inflammation, the anti-inflammatory effect of 7.5, 15, 30, and 60 μmol/L quercetin, asiaticoside, and asiatic acid, which were identified as the main active components in Centella asiatica, were tested by measuring cellular production of NO, TNF‑α and IL-6 using Griess method and ELISA and by detecting mRNA expressions of IL-23, IL-17A, TNF-α and IL-6 and protein expressions of p-STAT3 (Tyr705) and p-STAT3 (Ser727) with RT-qPCR and Western blotting. Results A total of 139 targets of Centella asiatica and 4604 targets of psoriasis were obtained, and among them CASP3, EGFR, PTGS2, and ESR1 were identified as the core targets. KEGG analysis suggested that quercetin, asiaticoside, and asiatic acid in Centella asiatica were involved in cancer and IL-17 and MAPK signaling pathways. In the RAW264.7 macrophage model of inflammation, treatment with quercetin significantly reduced cellular production of NO, TNF‑α and IL-6, and lowered mRNA expressions of IL-23, IL-17A, TNF‑α and IL-6 and protein expressions of p-STAT3 (Tyr705) and p-STAT3 (Ser727). Conclusion Quercetin, asiaticoside and asiatic acid are the main active components in Centella asiatica to mediate the therapeutic effect against psoriasis, and quercetin in particular is capable of suppressing cellular production of NO, TNF‑α and IL-6 and regulating the IL-23/IL-17A inflammatory axis by mediating STAT3 phosphorylation to inhibit inflammatory response.

Key words: Centella asiatica, psoriasis, quercetin, IL-23/IL-17A, network pharmacology

刘青, 刘敬, 郑逸航, 雷金, 黄建华, 刘思妤, 刘芳, 彭群龙, 张远芳, 王俊杰, 李玉娟. 积雪草活性成分槲皮素通过介导STAT3磷酸化抑制IL-23/IL-17A炎症轴发挥抗银屑病作用[J]. 南方医科大学学报, 2025, 45(1): 90-99.

Qing LIU, Jing LIU, Yihang ZHENG, Jin LEI, Jianhua HUANG, Siyu LIU, Fang LIU, Qunlong PENG, Yuanfang ZHANG, Junjie WANG, Yujuan LI. Quercetin mediates the therapeutic effect of Centella asiatica on psoriasis by regulating STAT3 phosphorylation to inhibit the IL-23/IL-17A axis[J]. Journal of Southern Medical University, 2025, 45(1): 90-99.

图/表 10

表1 实时荧光定量PCR引物序列

Tab.1 Primer sequences for real-time fluorescence quantitative PCR

Gene	Primer sequences (5'-3')-F	Primer sequences (5'-3')-R	Length (bp)
GAPDH	AGGTCGGTGTGAACGGATTTG	TGTAGACCATGTAGTTGAGGTCA	123
IL-23	GTGGCATCGAGAAACTGT	GAGCCACCCAGGAAAGTA	116
IL-17A	ACTACCTCAACCGTTCCACG	TTCCTCCGCATTGACACAG	120
TNF-α	ATGAGCACAGAAAGCATGATC	GGTCTGGGCCATAGAACTGATG	231
IL-6	TAGTCCTTCCTACCCCAATTTCC	TTGGTCCTTAGCCACTCCTTC	76

表2 积雪草活性成分基本信息

Tab.2 Basic information of the active components in Centella asiatica

Number	MOL ID	Compound Name	Oral Bioavailability	Drug Likeness
A 1	MOL000098	Quercetin	46.43	0.28
A 2	MOL007312	Asiaticoside	10.22	0.7
A 3	MOL007253	Asiatic acid	16.69	0.72
A 4	MOL000359	Sitosterol	36.91	0.75
A 5	MOL007313	Xanthanoic acid	48.07	0.16
A 6	MOL007320	8-acetoxycentellynol	65.94	0.12
A 7	MOL006370	5-O-caffeoylquinic acid	19.61	0.33
A 8	MOL000008	Qpigenin	23.06	0.21
A 9	MOL007201	Brahmic acid	17.6	0.7
A 10	MOL006387	Chlorogenic acid	25.58	0.33
A 11	MOL007323	Madasiatic acid	18.42	0.72
A 12	MOL007303	Madecassoside	16.89	0.7
A 13	MOL006407	Neochlorogenic acid	18.05	0.33
A 14	MOL001434	Quercetin 3-O-rhamnopyranosyl	22.24	0.28
A 15	MOL000511	Ursolic acid	16.77	0.75

图 1 积雪草活性成分-银屑病靶点韦恩图

Fig. 1 Venn diagram of the active components of Centella asiatica-psoriasis target.

图2 "药物-活性成分-作用靶点"和PPI网络图

Fig.2 Drug-active ingredient-targets and PPI network diagram. A: Drug-active ingredient-targets network diagram. B: PPI network diagram. C: Core targets network diagram.

图3 积雪草抗银屑病GO功能注释与KEGG通路分析

Fig. 3 GO functional annotation and KEGG signal pathway analysis of the anti-psoriasis effect of Centella asiatica. A: GO functional annotation. B: KEGG signal pathway analysis.

图 4 积雪草活性成分对RAW264.7细胞活力的影响

Fig.4 Effect of active components in Centella asiatica on the growth of RAW264.7 cells. A-C: Effects of Que, Asi, and Asi acid on viability of RAW264.7 cells. D-F: Effects of Que, Asi, and Asi acid on viability of RAW264.7 cells treated with LPS (100 ng/mL). *P<0.05, **P<0.01 vs Control group.

图 5 积雪草活性成分对LPS诱导的RAW264.7细胞产生NO的影响

Fig. 5 Effect of Que (A), Asi (B) and Asi acid (C) on NO levels in the cell culture media of LPS-induced RAW264.7 cells determined by Griess assay. ##P<0.01 vs Control group, *P<0.05, **P<0.01 vs LPS group.

图 6 槲皮素对细胞促炎因子含量的影响

Fig. 6 Inhibitory effects of Que on secretion of TNF-α (A) and IL-6 (B) in LPS-induced RAW264.7 cells. ##P<0.01 vs Control group, *P<0.05, **P<0.01 vs LPS group.

图 7 槲皮素对炎症细胞中IL-23、IL-17A、TNF-α和IL-6 mRNA表达的影响

Fig. 7 Inhibitory effects of Que on mRNA expressions of IL-23 (A), IL-17A (B), TNF-α (C), and IL-6 (D) in LPS-induced RAW264.7 cells. ##P<0.01 vs Control group, *P<0.05, **P<0.01 vs LPS group.

图 8 槲皮素对炎症细胞中STAT3蛋白磷酸化影响

Fig. 8 Effect of Que on STAT3 protein phosphorylation in LPS-induced RAW264.7 cells. A: Western blotting for detecting expressions of STAT3, p-STAT3 (Tyr705), and p-STAT3 (Ser727). B-D: Semi-quantitative analysis of STAT3, p-STAT3(Tyr705), and p-STAT3(Ser727) expression levels. ##P<0.01 vs Control group, *P<0.05, **P<0.01 vs LPS group.

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积雪草活性成分槲皮素通过介导STAT3磷酸化抑制IL-23/IL-17A炎症轴发挥抗银屑病作用

Quercetin mediates the therapeutic effect of Centella asiatica on psoriasis by regulating STAT3 phosphorylation to inhibit the IL-23/IL-17A axis

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