基于HPLC指纹图谱和网络药理学的白屈菜缓解IPEC-J2细胞炎性损伤作用研究

doi:10.11843/j.issn.0366-6964.2025.05.042

Abstract

Abstract:

Chelidonium majus can be used to treat various digestive and respiratory tract inflammations, but its mechanism of action is still unclear. This study predicts and analyzes the pharmacodynamic components and mechanisms of C. majus using fingerprint and network pharmacology. A total of 15 batches of C. majus fingerprint were established using HPLC and evaluated for similarity and principal component analysis. A network of active components and targets of C. majus was established using network pharmacology methods. The core anti-inflammatory components were analyzed, along with the core targets through protein interaction network analysis, followed by GO enrichment and KEGG enrichment analysis. The common peaks identified by fingerprint and the core components filtered by network pharmacology were intersected. The effects of C. majus on IPEC-J2 cell inflammation damage were studied using MTT and ELISA methods. The results showed that 12 out of 15 batches of C. majus had a similarity >0.9, with 21 common peaks identified, among which 6 were identified as protopine, chelidonine, coptisine, sanguinarine, berberine, and chelerythrine. Network pharmacology filtered out 8 core anti-inflammatory components, namely thalictriline, (s)-thalictroidine, (s)-tetrahydropalmatine, berberine, chelidonine, chelerythrine, oxidized sanguinarine, and (+/-)-thalicarpine. The intersection of these two sets of components was chelidonine, berberine, and chelerythrine. Referring to the regulations of the Chinese Pharmacopoeia, chelerythrine was selected for a study on its effect on IPEC-J2 cell inflammation. The study found that chelerythrine at concentrations of 2.5 to 10 μg·mL^-1 could significantly improve cell survival rate, increase the content of IL-4 and IL-10, and decrease the content of NO, IL-1β, IL-6, IL-8, TNF-α, and TGF-β1. This study has established the HPLC fingerprint of C. majus and preliminarily revealed its pharmacodynamic components and mechanisms of action using network pharmacology methods. This provides a reference for its quality control and further development.

Key words: Chelidonium majus, fingerprint, network pharmacology, chelerythrine, anti-inflammatory

CLC Number:

S853.74

CHEN Zehan, ZHANG Ruoyi, LIN Huiying, ZENG Chunli, LIN Fu, LI Jian. Anti-inflammatory Effects of Chelidonium majus on IPEC-J2 Cells based on HPLC Fingerprint and Network Pharmacology[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(5): 2466-2480.

Figures/Tables 14

Table 1

Fig. 1

Table 2

Table 3

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Fig. 6

Fig. 7

Fig. 8

Fig. 9

Fig. 10

Fig. 11

References 35

1	ZIELIŃSKA S , JEZIERSKA-DOMARADZKA A , WÓJCIAK-KOSIOR M , et al. Greater celandine's ups and downs-21 centuries of medicinal uses of Chelidonium majus from the viewpoint of today's pharmacology[J]. Front Pharmacol, 2018, 9, 299. doi: 10.3389/fphar.2018.00299
2	朱橚(明). 救荒本草校注[M]. 北京: 中国农业出版社, 2008.
	ZHU S. Translation and annotation of Jiuhuang Bencao or treatise on wild food plants used for saving famine[M]. Beijing: China Agriculture Press, 2008. (in Chinese)
3	国家药典委员会. 中华人民共和国药典(一部)[M]. 北京: 中国医药科技出版社, 2020.
	Chinese Pharmacopoeia Commission . Pharmacopoeia of the People's Republic of China[M]. Beijing: China Pharmaceutical Science and Technology Press, 2020.
4	中国兽药典委员会. 中华人民共和国兽药典(二部)[M]. 北京: 中国农业出版社, 2020.
	Veterinary Pharmacopoeia Committee of China . Pharmacopoeia of the People's Republic of China (Volume Ⅱ)[M]. Beijing: China Agriculture Press, 2020.
5	朱静琳. 白屈菜靶向肿瘤微环境腺苷信号通路的抗肿瘤机制研究[D]. 西安: 西北大学, 2022.
	ZHU J L. Antitumor mechanism of Chelidonium majus L. targeting adenosine signaling pathway within the tumor microenvironment[D]. Xi'an: Northwest University, 2022. (in Chinese)
6	刘金龙. 白屈菜红碱对三阴性乳腺癌抑制作用及机制研究[D]. 长春: 长春中医药大学, 2022.
	LIU J L. Study on the inhibitory effect and mechanism of chelerythrine on triple-negative breast cancer[D]. Changchun: Changchun University of Chinese Medicine, 2022. (in Chinese)
7	江健梅. 驯化栽培白屈菜质量评价[D]. 长春: 吉林农业大学, 2017.
	JIANG J M. Study on quality evaluation of demesticated Chelidonium majus L. [D]. Changchun: Jilin Agricultural University, 2017. (in Chinese)
8	林惠莹, 赵永兵, 李荣强, 等. 白屈菜药理作用及质量标志物预测分析[J]. 中兽医医药杂志, 2022, 41 (2): 41- 45.
	LIN H Y , ZHAO Y B , LI R Q , et al. Pharmacological activities of Chelidonii herba and predictive analysis on Q-marker[J]. Journal of Traditional Chinese Veterinary Medicine, 2022, 41 (2): 41- 45.
9	林惠莹, 杨黎宇, 李家伟, 等. 白屈菜碱对H₂O₂诱导的IPEC-J2细胞炎症损伤的预防作用[J]. 中国兽医学报, 2024, 44 (1): 128- 134.
	LIN H Y , YANG L Y , LI J W , et al. Preventive effect of chelidonine on H₂O₂-induced inflammatory injury of IPEC-J2 cells[J]. Chinese Journal of Veterinary Science, 2024, 44 (1): 128- 134.
10	王倩, 姜俊, 许霞, 等. 指纹图谱及网络药理学联合分析芪胶升白胶囊的药效成分及机制[J]. 中国中药杂志, 2023, 48 (6): 1526- 1534.
	WANG Q , JIANG J , XU X , et al. Effective components and mechanism of Qijiao Shengbai Capsules based on fingerprinting and network pharmacology[J]. China Journal of Chinese Materia Medica, 2023, 48 (6): 1526- 1534.
11	王迎春, 马永犇, 甄亚钦, 等. 基于指纹图谱和网络药理学的芍药甘草汤抗肝损伤活性成分及含量测定研究[J]. 世界科学技术-中医药现代化, 2022, 24 (8): 3030- 3042.
	WANG Y C , MA Y B , ZHEN Y Q , et al. Integrated fingerprint and network pharmacology approaches to explore the anti-liver injury active constituents and content determination of shaoyao gancao decoction[J]. Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology, 2022, 24 (8): 3030- 3042.
12	孙媛, 王璐, 彭梅梅, 等. 甘草不同炮制品的指纹图谱研究及质量标志物预测分析[J]. 中国中药杂志, 2020, 45 (21): 5209- 5218.
	SUN Y , WANG L , PENG M M , et al. Fingerprint analysis and Q-marker prediction of processed liquorice products[J]. China Journal of Chinese Materia Medica, 2020, 45 (21): 5209- 5218.
13	LIU Q , LIU L Y , XIE L T , et al. Screening and evaluation of quality markers of Radix Cudramiae for liver disease based on an integrated strategy of in vivo pharmacokinetics and in vitro HPLC fingerprint[J]. J Pharm Biomed Anal, 2024, 242, 116055. doi: 10.1016/j.jpba.2024.116055
14	SHEN C J , WANG J , FENG M J , et al. The mitochondrial-derived peptide MOTS-c attenuates oxidative stress injury and the inflammatory response of H9C2 cells through the Nrf2/ARE and NF-κB pathways[J]. Cardiovasc Eng Technol, 2022, 13 (5): 651- 661. doi: 10.1007/s13239-021-00589-w
15	杨明慧. 吉林省不同产地白屈菜有效成分含量及药效比较[D]. 长春: 长春中医药大学, 2019.
	YANG M H. Content and pharmacodynamic comparison of active components of chelidonium majus from different origins in Jilin Province[D]. Changchun: Changchun University of Chinese Medicine, 2019. (in Chinese)
16	孟琦. 野生白屈菜主要有效成分积累动态及影响因子研究[D]. 长春: 吉林农业大学, 2017.
	MENG Q. Studies on the dynamic changes of chemical components and the influencing factors of wild Chelidonium majus L. [D]. Changchun: Jilin Agricultural University, 2017. (in Chinese)
17	李晓蒙, 朱盛华, 廖华卫. 白屈菜药材HPLC指纹图谱研究[J]. 天然产物研究与开发, 2002, 14 (6): 33- 37. doi: 10.3969/j.issn.1001-6880.2002.06.012
	LI X M , ZHU S H , LIAO H W . The HPLC fingerprint of Chelidonium majus L.[J]. Natural Product Research and Development, 2002, 14 (6): 33- 37. doi: 10.3969/j.issn.1001-6880.2002.06.012
18	TERZIC M , FAYEZ S , FAHMY N M , et al. Chemical characterization of three different extracts obtained from Chelidonium majus L. (Greater celandine) with insights into their in vitro, in silico and network pharmacological properties[J]. Fitoterapia, 2024, 174, 105835. doi: 10.1016/j.fitote.2024.105835
19	陈思睿, 吴天鸿, 刘洁, 等. 基于网络药理学、分子对接、实验研究探讨白屈菜治疗鼻咽癌的物质基础及潜在机制[J]. 湖南中医药大学学报, 2024, 44 (2): 278- 287.
	CHEN S R , WU T H , LIU J , et al. Material basis and potential mechanism of Baiqucai (Chelidonii Herba) in treating nasopharyngeal carcinoma based on network pharmacology, molecular docking, and experimental research[J]. Journal of Hunan University of Chinese Medicine, 2024, 44 (2): 278- 287.
20	张蕾, 王敏, 张新新, 等. 基于网络药理学和分子对接技术分析白屈菜红碱抗乳腺癌的作用机制[J]. 西安交通大学学报: 医学版, 2021, 42 (4): 554-561, 573.
	ZHANG L , WANG M , ZHANG X X , et al. The mechanism of chelerythrine against breast cancer by network pharmacology and molecular docking[J]. Journal of Xi'an Jiaotong University: Medical Sciences, 2021, 42 (4): 554-561, 573.
21	章亮, 陈泽慧, 陈韩英, 等. 基于网络药理学的白屈菜抗肿瘤分子机制研究[J]. 中草药, 2018, 49 (3): 646- 657.
	ZHANG L , CHEN Z H , CHEN H Y , et al. Study on antitumor molecular mechanism of Chelidonium majus based on network pharmacology[J]. Chinese Traditional and Herbal Drugs, 2018, 49 (3): 646- 657.
22	SUN Q Y , ZHOU H H , MAO X Y . Emerging Roles of 5-Lipoxygenase phosphorylation in inflammation and cell death[J]. Oxid Med Cell Longev, 2019, 2019, 2749173.
23	COPE A P . Studies of T-cell activation in chronic inflammation[J]. Arthritis Res Ther, 2002, 4 (3): S197.
24	KOLB J P , OGUIN T H , OBERST A , et al. Programmed cell death and inflammation: winter is coming[J]. Trends Immunol, 2017, 38 (10): 705- 718. doi: 10.1016/j.it.2017.06.009
25	LOCK C B , HELLER R A . Gene microarray analysis of multiple sclerosis lesions[J]. Trends Mol Med, 2003, 9 (12): 535- 541. doi: 10.1016/j.molmed.2003.10.008
26	YASUDA K , TAKEUCHI Y , HIROTA K . The pathogenicity of Th17 cells in autoimmune diseases[J]. Semin Immunopathol, 2019, 41 (3): 283- 297. doi: 10.1007/s00281-019-00733-8
27	BAILEY A S , ROBINSON R . Synthesis of a substance containing the chelerythrine-sanguinarine skeleton (C, N, O)[J]. Nature, 1949, 164 (4166): 402.
28	KOSINA P , VACEK J , PAPOUŠKOVÁ B , et al. Identification of benzo[c]phenanthridine metabolites in human hepatocytes by liquid chromatography with electrospray ion-trap and quadrupole time-of-flight mass spectrometry[J]. J Chromatogr B, 2011, 879 (15-16): 1077- 1085. doi: 10.1016/j.jchromb.2011.03.023
29	周佳, 邱智东, 王雨辰, 等. 白屈菜红碱的研究进展[J]. 人参研究, 2022, 34 (6): 46- 49.
	ZHOU J , QIU Z D , WANG Y C , et al. Research progress on chelerythrine[J]. Ginseng Research, 2022, 34 (6): 46- 49.
30	苑庆欣, 刘阔, 包旭华, 等. 白屈菜红碱抗耐甲氧西林金黄色葡萄球菌作用机制研究[J]. 畜牧兽医学报, 2024, 55 (10): 4670- 4678. doi: 10.11843/j.issn.0366-6964.2024.10.038
	YUAN Q X , LIU K , BAO X H , et al. The mechanism of chelerythrine against methicillin-resistant Staphylococcus aureus[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55 (10): 4670- 4678. doi: 10.11843/j.issn.0366-6964.2024.10.038
31	方伟燕. 补脾益肠丸干预IL-21/IL-21R调控记忆性T细胞分化治疗溃疡性结肠炎的作用机制[D]. 南昌: 江西中医药大学, 2022.
	FANG W Y. Effect of Bupi Yichang Pill treated ulcerative colitis by regulating memory T cell differentiation via IL-21/IL-21R[D]. Nanchang: Jiangxi University of Traditional Chinese Medicine, 2022. (in Chinese)
32	LI L , CHEN L , LIN F , et al. Study of the expression of inflammatory factors IL-4, IL-6, IL-10, and IL-17 in liver failure complicated by coagulation dysfunction and sepsis[J]. J Inflamm Res, 2021, 14, 1447- 1453. doi: 10.2147/JIR.S302975
33	MANTOVANI A , DINARELLO C A , MOLGORA M , et al. Interleukin-1 and related cytokines in the regulation of inflammation and immunity[J]. Immunity, 2019, 50 (4): 778- 795. doi: 10.1016/j.immuni.2019.03.012
34	CAMPOREALE A , POLI V . IL-6, IL-17 and STAT3: a holy trinity in auto-immunity?[J]. Front Biosci (Landmark Ed), 2012, 17 (6): 2306- 2326.
35	SIEPER J , PODDUBNYY D , MIOSSEC P . The IL-23-IL-17 pathway as a therapeutic target in axial spondyloarthritis[J]. Nat Rev Rheumatol, 2019, 15 (12): 747- 757. doi: 10.1038/s41584-019-0294-7

编号 No.	产地 Place of production	编号 No.	产地 Place of production	编号 No.	产地 Place of production
S1	辽宁	S6	江西	S11	云南
S2	吉林	S7	浙江	S12	安徽
S3	河北	S8	四川	S13	广西
S4	河南	S9	陕西	S14	湖北
S5	江苏	S10	黑龙江	S15	福建

成分Ingredient	回归方程Regression equation	r²
原阿片碱Protopine	y=13 796x+4 626.2	0.999 3
白屈菜碱Chelidonine	y =4 476.4x+6 751.9	0.999 0
黄连碱Coptisine	y=22 512x+7 186.9	0.999 8
血根碱Sanguinarine	y=29 031x-1 564.4	0.999 9
小檗碱Berberine	y=23 151x+5.896	0.999 8
白屈菜红碱 Chelerythrine	y=54 706x-1 948	0.999 9

编号 No.	S1	S2	S3	S4	S5	S6	S7	S8
S1	1.000	0.981	0.995	0.964	0.991	0.973	0.990	0.691
S2	0.981	1.000	0.987	0.986	0.990	0.987	0.989	0.740
S3	0.995	0.987	1.000	0.975	0.997	0.981	0.998	0.724
S4	0.964	0.986	0.975	1.000	0.982	0.998	0.981	0.762
S5	0.991	0.990	0.997	0.982	1.000	0.984	0.999	0.734
S6	0.973	0.987	0.991	0.998	0.994	1.000	0.984	0.752
S7	0.990	0.931	0.999	0.930	0.997	0.983	1.000	0.106
S8	0.691	0.143	0.071	0.167	0.106	0.147	0.106	1.000
S9	0.554	0.631	0.585	0.677	0.601	0.663	0.603	0.946
S10	0.179	0.211	0.194	0.241	0.191	0.242	0.196	0.750
S11	0.993	0.987	0.994	0.982	0.996	0.986	0.994	0.721
S12	0.971	0.987	0.980	0.998	0.984	0.999	0.984	0.755
S13	0.980	0.992	0.987	0.965	0.993	0.995	0.992	0.746
S14	0.986	0.994	0.992	0.992	0.996	0.993	0.995	0.742
S15	0.987	0.992	0.991	0.993	0.994	0.996	0.993	0.737

编号 No.	S9	S10	S11	S12	S13	S14	S15	对照指纹图谱 Control fingerprint
S1	0.554	0.179	0.993	0.971	0.980	0.986	0.987	0.968
S2	0.631	0.211	0.987	0.987	0.992	0.994	0.992	0.983
S3	0.585	0.194	0.994	0.980	0.987	0.992	0.991	0.979
S4	0.677	0.241	0.982	0.998	0.995	0.992	0.993	0.987
S5	0.601	0.191	0.996	0.984	0.993	0.996	0.994	0.982
S6	0.663	0.242	0.986	0.999	0.995	0.993	0.996	0.988
S7	0.043	0.995	0.910	0.984	0.992	0.995	0.993	0.983
S8	0.389	0.105	0.176	0.755	0.746	0.742	0.737	0.832
S9	1.000	0.817	0.596	0.666	0.645	0.632	0.631	0.734
S10	0.817	1.000	0.194	0.245	0.211	0.208	0.209	0.354
S11	0.596	0.194	1.000	0.985	0.991	0.995	0.994	0.980
S12	0.666	0.245	0.985	1.000	0.995	0.993	0.995	0.988
S13	0.645	0.211	0.991	0.995	1.000	0.998	0.999	0.987
S14	0.632	0.208	0.995	0.993	0.998	1.000	0.998	0.988
S15	0.631	0.209	0.994	0.995	0.999	0.998	1.000	0.987

[1]	Ling LIU, Wanyu SHI, Xiumei LI, Minghua WANG, Xianghe ZHAI, Weiwei ZHOU. Intervention of Essential Oils from Citrus reticulata Blanco on Lipopolysaccharide- induced Inflammation in RAW264.7 Cells [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(9): 4153-4160.
[2]	Anlin WEN, Yunyun YANG, Yongrong LUO, Ying YANG, Zhentao CHENG, Deyuan OU, Ming WEN. Network Pharmacologic Analysis and Experimential Verification on the Mechanism of Coptidis Rhizoma in Preventing Duck Viral Enteritis [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(7): 3225-3233.
[3]	Chengquan HAN, Jingtao LIU, Miao YU, Lizeng GUAN, Lu XU, Yueshang WANG. Study on the Anti-Toxoplasma gondii Effect of Rutin in BV2 Cells and Its Mechanism [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(7): 3119-3131.
[4]	XU Junjie, ZHANG Lutong, WANG Jinjie, CHEN Xiaochen, HE Weixian, CAI Chuanjiang, CHU Guiyan, YANG Gongshe. Exploring the Effect of Epimedium on Estrus of Gilts Based on Multiomics and Network Pharmacology [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(4): 1615-1628.
[5]	LIU Yuanhong, HU Yuhuan, ZHANG Li, YANG Pingrui, HU Weidong, MA Qi, BI Shicheng. Network Pharmacologic Analysis and Experimental Verification of Atractylodes Macrocephala-Cistanche Deserticola in the Treatment of Constipation [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(2): 834-845.
[6]	TIAN Yan, YANG Fusheng, LI Yan'e, LIANG Youping, FAN Jie, WU Fangyan, GU Xiaobin. Effect of Recombinant Serpins of Psoroptes ovis on Psoriasis-like Skin Inflammation in Mice [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5774-5783.
[7]	Qingxin YUAN, Kuo LIU, Xuhua BAO, Dongyang GAO, He LI, Jun SONG, Zhixin ZHOU. The Mechanism of Chelerythrine against Methicillin-resistant Staphylococcus aureus [J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(10): 4670-4678.
[8]	CHEN Xihong, LU Guicong, WANG Haolei, GOU Shaoxiao, YU Yongxiong, LIN Tao, JIANG Caode. Isochlorogenic Acid C Inhibits Mammary Inflammatory Response through NF-κB Signaling Pathway Using Bovine Mammary Gland Cells and Mouse Mammary Gland Tissue [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3931-3940.
[9]	ZHANG Xumei, WEI Yurong, XU Chenghui, YANG Tong, SHI Huijun, FU Qiang, YANG Li. To Analyze the Mechanism of Berberine in the Treatment of Salmonella Gallinarum Infection Based on Network Pharmacology and Experimental Verification [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3557-3570.
[10]	GONG Zhiguo, ZHAO Jiamin, GU Baichen, REN Peipei, YU Zhuoya, BAI Yunjie, LIU Xinyu, WANG Chao, LIU Bo. Exploring the Mechanism of Codonopsis pilosula in Alleviation of Acute Lung Injury in Escherichia coli Infected Mice Based on Network Pharmacology [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(8): 3571-3581.
[11]	YANG Zihui, DONG Zhen, WU Huilan, TAN Bin, ZENG Jianguo. Analysis of the Material Basis and Mechanism of Action of Antioxidant Function of Taraxacum mongolicum based on Network Pharmacology [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2170-2185.
[12]	PAN Chanyuan, ZHAO Zixuan, DUAN Mingjie, JIANG Linshu, TONG Jinjin. The Mechanism of Artemisia carvifolia Alleviating Dairy Cow Oxidative Stress Predicted by Network Pharmacology [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1071-1084.
[13]	SUN Panpan, CAO Zhigang, LING Xiaoya, SUN Na, SUN Yaogui, LI Hongquan. Comparison of Anti-inflammatory Effects of Matrine Combined with Different Antibiotics [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(10): 4411-4421.
[14]	CUI Enhui, XUE Yuhuan, LI Cixia, WANG Shuai, ZHU Xiaoyan, CHAI Xuejun, ZHAO Shanting. Network Pharmacological Analysis and Experimental Verification of Immunomodulatory Mechanisms of Eucommia ulmoides Leaf [J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(1): 403-413.
[15]	TIAN Wei, LI Xiumei, YANG Juan, WANG Xiaoying, ZHOU Weiwei, LI Zhongyuan, DAI Xiaofeng. Network Pharmacology Study on the Antibacterial Activity Components and Mechanism of Isatis indigotica [J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(8): 2782-2793.

Anti-inflammatory Effects of Chelidonium majus on IPEC-J2 Cells based on HPLC Fingerprint and Network Pharmacology

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 14

References 35

Related Articles 15

Recommended Articles

Metrics

Comments