阔叶十大功劳抗炎作用机制的实验研究及网络药理学分析

doi:10.3976/j.issn.1002-4026.20230043

摘要/Abstract

摘要：

基于网络药理学及动物实验研究苗药阔叶十大功劳抗炎的有效成分及作用机制。用二甲苯、角叉菜胶分别制作小鼠及大鼠的炎症模型,并灌胃阔叶十大功劳(小鼠剂量为1.950 mg/kg;大鼠剂量为1.350 mg/kg)。通过TCMSP、SymMap、SwisstargetPrediction、SEA、STICH等数据库,以口服生物利用度≥30%、类药性≥0.18获取阔叶十大功劳的活性成分及其相应靶点;通过GeneCards、DisGeNET、TTD、DrugBank、OMIM等数据库获取炎症相关靶点;通过Venny 2.1.0获取疾病和药物的交集靶点,通过Cytoscape的MCODE、CytoHubba插件得到10个关键靶点,并构建药物-成分-靶点网络图;对10个Hub gene进行KEGG(Kyoto encyclopedia of genes and genomes )和GO (gene ontology)富集分析以及分子对接。动物实验结果表明,阔叶十大功劳能够减轻小鼠和大鼠的炎症症状。网络药理学分析获得28个阔叶十大功劳活性化合物,753个药物作用靶点,1 025个炎症靶点,阔叶十大功劳-炎症交集靶点225个,Hub gene 10个。GO及 KEGG富集分析结果显示,阔叶十大功劳主要参与JAK-STAT信号传导途径、Th17细胞分化和一些癌症通路。分子对接结果显示,小檗碱、异博尔定等11个活性成分与JUN、JAK3等8个靶点对接成功。动物实验结果表明阔叶十大功劳具有抗炎作用,抗炎的主要成分为槲皮素、小檗碱等化合物,抗炎的机制可能是通过作用于IL-2、JAK1等靶点,参与JAK-STAT信号传导途径、Th17细胞分化等通路抗炎,初步揭示了阔叶十大功劳发挥抗炎作用的物质基础及作用机制。

关键词: 阔叶十大功劳, 网络药理学, 炎症, 分子机制, 有效成分

Abstract:

The aim of this study was to investigate the active ingredients and mechanism of action of the anti-inflammatory effect of Mahonia bealei (Fort.) Carr. in Hmong medicine based on network pharmacology and animal experiments. Inflammation models of mice and rats were generated using xylene and carrageenan gum, respectively, and Mahonia bealei (Fort.) Carr. was gavaged (dose of 1.950 mg/kg for mice; 1.350 mg/kg for rats). The active ingredients and their corresponding targets of Mahonia bealei (Fort.) Carr. were obtained using TCMSP, SymMap, SwisstargetPrediction, SEA, and STICH, among other databases, with oral bioavailability ≥30% and drug-likeness ≥0.18. Inflammationrelated targets were obtained through GeneCards, DisGeNET, TTD, DrugBank, OMIM and other databases. The intersection targets of diseases and drugs were determined using Venny 2.1.0, and 10 hub gene were obtained through Cytoscape's MCODE, CytoHubba plug-in and constructed drug-component-target network diagram; Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Ontology (GO) enrichment analysis and molecular docking were performed for the 10 Hub gene. The results of animal experiments showed that Mahonia bealei (Fort.) Carr. could be used to reduce the inflammatory symptoms in mice and rats. The network pharmacological analysis revealed 28 Mahonia bealei (Fort.) Carr. active ingredients, 753 drug action targets, 1 025 inflammatory targets, 225 Mahonia bealei (Fort.) Carr. inflammatory crossover targets and 10 hub genes. The results of GO and KEGG enrichment analysis showed that Mahonia bealei (Fort.) Carr. top ten utilities were predominantly involved in the JAK-STAT signaling pathway, Th17 cell differentiation and some cancer pathways. The molecular docking results demonstrated that 11 active ingredients, including berberine and isoboridine, were successfully docked with 8 targets, including JUN and JAK3. The results of animal experiments showed that Mahonia bealei (Fort.) Carr. has anti-inflammatory effects, and the main ingredients of anti-inflammation include quercetin and berberine, among other compounds, and the mechanism of anti-inflammation may be through the action onIL-2, JAK1 and other targets, involved in JAK-STAT signaling pathway, Th17 cell differentiation, and other pathways of anti-inflammation. The present study initially revealed the material basis and mechanism of action of the anti-inflammatory effect of Mahonia bealei (Fort.) Carr.

Key words: Mahonia bealei (Fort.) Carr., network pharmacology, inflammation, molecular mechanism, active ingredient

中图分类号:

R285

付朝举, 成果, 林登梅, 李军, 张楠楠. 阔叶十大功劳抗炎作用机制的实验研究及网络药理学分析[J]. 山东科学, 2024, 37(1): 24-31.

FU Zhaoju, CHENG Guo, LIN Dengmei, LI Jun, ZHANG Nannan. Experimental study and network pharmacological analysis of the anti-inflammatory action mechanism of Mahonia bealei (Fort.) Carr.[J]. Shandong Science, 2024, 37(1): 24-31.

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参考文献 25

[1]	中国科学院中国植物志编辑委员会. 中国植物志第二十九卷[M]. 北京: 科学出版社, 2001: 235.
[2]	汪毅. 中国苗族药物彩色图集[M]. 贵阳: 贵州科技出版社, 2002.
[3]	贵州省民委文教处. 苗族医药学[M]. 贵阳: 贵州民族出版社, 1992.
[4]	刘偲翔, 刘布鸣, 何开家, 等. 两种十大功劳叶中生物碱成分的分析与比较[J]. 中国药业, 2010, 19(5): 4-5. DOI: 10.3969/j.issn.1006-4931.2010.05.002.
[5]	郭立敏, 吕洁丽, 张来宾. 天然倍半萜类化合物抗炎作用机制的研究进展[J]. 中国中药杂志, 2018, 43(20): 3989-3999. DOI: 10.19540/j.cnki.cjcmm.20180726.013.
[6]	KIM H J, JOE H I, ZHANG Z Y, et al. Anti-inflammatory effect of Acalypha australis L. via suppression of NF-κB signaling in LPS-stimulated RAW 264.7 macrophages and LPS-induced septic mice[J]. Molecular Immunology, 2020, 119: 123-131. DOI: 10.1016/j.molimm.2020.01.010.
[7]	LI S S, LI J, SUN J, et al. Berkeleyacetal C, a meroterpenoid isolated from the fungus Penicillium purpurogenum MHZ 111, exerts anti-inflammatory effects via inhibiting NF-κB, ERK1/2 and IRF3 signaling pathways[J]. European Journal of Pharmacology, 2017, 814: 283-293. DOI: 10.1016/j.ejphar.2017.08.039.
[8]	MITTEER D R, GREER B D, RANDALL K R, et al. Further evaluation of teaching behavior technicians to input data and graph using GraphPad Prism[J]. Behavior Analysis: Research and Practice, 2020, 20(2): 81-93. DOI: 10.1037/bar0000172.
[9]	杨翰林, 罗川晋, 吴伟. 湿热与瘀热类炎症的中医思考[J]. 中西医结合心脑血管病杂志, 2019, 17(3): 464-467. DOI: 10.12102/j.issn.1672-1349.2019.03.042.
[10]	YE C X, BRAND D, ZHENG S G. Targeting IL-2: an unexpected effect in treating immunological diseases[J]. Signal Transduction and Targeted Therapy, 2018, 3: 2. DOI: 10.1038/s41392-017-0002-5. pmid: 29527328
[11]	缪志伟, 徐艳, 宁丽琴, 等. 白头翁汤治疗溃疡性结肠炎分子机制的网络药理学分析及初步验证[J]. 中国中药杂志, 2020, 45(8): 1808-1815. DOI: 10.19540/j.cnki.cjcmm.20191112.406.
[12]	KIM D K, LIM J H, LEE J, et al. Synthesis and biological evaluation of trisubstituted imidazole derivatives as inhibitors of p38α mitogen-activated protein kinase[J]. Bioorganic & Medicinal Chemistry Letters, 2008, 18(14): 4006-4010. DOI: 10.1016/j.bmcl.2008.06.007.
[13]	LI Y, ZENG R J, CHEN J Z, et al. Pharmacokinetics and metabolism study of isoboldine, a major bioactive component from Radix Linderae in male rats by UPLC-MS/MS[J]. Journal of Ethnopharmacology, 2015, 171: 154-160. DOI: 10.1016/j.jep.2015.05.042. pmid: 26055342
[14]	PARK H H, KIM S G, PARK Y N, et al. Suppressive effects of britanin, a sesquiterpene compound isolated from inulae Flos, on mast cell-mediated inflammatory responses[J]. The American Journal of Chinese Medicine, 2014, 42(4): 935-947. DOI: 10.1142/s0192415x14500591.
[15]	KIM S G, LEE E, PARK N Y, et al. Britanin attenuates ovalbumin-induced airway inflammation in a murine asthma model[J]. Archives of Pharmacal Research, 2016, 39(7): 1006-1012. DOI: 10.1007/s12272-016-0783-z.
[16]	LI C L, TAN L H, WANG Y F, et al. Comparison of anti-inflammatory effects of berberine, and its natural oxidative and reduced derivatives from Rhizoma Coptidis in vitro and in vivo[J]. Phytomedicine: International Journal of Phytotherapy and Phytopharmacology, 2019, 52: 272-283. DOI: 10.1016/j.phymed.2018.09.228.
[17]	李红枚, 党万太, 杨小红. 小檗碱抗炎机制的研究进展[J]. 中国医药导报, 2017, 14(33): 31-34.
[18]	CALICETI C, FRANCO P, SPINOZZI S, et al. Berberine: new insights from pharmacological aspects to clinical evidences in the management of metabolic disorders[J]. Current Medicinal Chemistry, 2016, 23(14): 1460-1476. DOI: 10.2174/0929867323666160411143314. pmid: 27063256
[19]	BYUN E B, YANG M S, CHOI H G, et al. Quercetin negatively regulates TLR4 signaling induced by lipopolysaccharide through Tollip expression[J]. Biochemical and Biophysical Research Communications, 2013, 431(4): 698-705. DOI: 10.1016/j.bbrc.2013.01.056.
[20]	范丽梅, 柳昀熠, 潘钰, 等. 炎症与肿瘤的关系研究进展[J]. 江汉大学学报(自然科学版), 2016, 44(5): 432-437. DOI: 10.16389/j.cnki.cn42-1737/n.2016.05.009.
[21]	BANERJEE S, BIEHL A, GADINA M, et al. JAK-STAT signaling as a target for inflammatory and autoimmune diseases: current and future prospects[J]. Drugs, 2017, 77(5): 521-546. DOI: 10.1007/s40265-017-0701-9. pmid: 28255960
[22]	DONG C. Differentiation and function of pro-inflammatory Th17 cells[J]. Microbes and Infection, 2009, 11(5): 584-588. DOI: 10.1016/j.micinf.2009.04.001. pmid: 19371793
[23]	MOSS R B, MOLL T, EL-KALAY M, et al. Th1/Th2 cells in inflammatory disease states: Therapeutic implications[J]. Expert Opinion on Biological Therapy, 2004, 4(12): 1887-1896. DOI: 10.1517/14712598.4.12.1887.
[24]	ROMAGNANI S. Th1/Th2 cells[J]. Inflammatory Bowel Diseases, 1999, 5(4): 285-294. DOI: 10.1097/00054725-199911000-00009. pmid: 10579123
[25]	WALKER J A, MCKENZIE A N J. TH2 cell development and function[J]. Nature Reviews Immunology, 2018, 18(2): 121-133. DOI: 10.1038/nri.2017.118.

Tukey的多重比较试验	平均差异	95.00% CI of diff	Summary	P值
空白组vs.阳性组	0.416 5	0.143 4~0.689 5	**	0.003 4
空白组vs.实验组	0.418 8	0.145 8~0.691 9	**	0.003 2
阳性组vs.实验组	0.002 338	-0.270 7~0.275 4	ns	0.999 7

Tukey的多重比较试验	平均差异	95.00% CI of diff	Summary	P值
空白组vs.阳性组	0.336 9	0.131 5~0.542 2	**	0.001 9
空白组vs.实验组	0.230 1	0.024 81~0.435 5	*	0.027 3
阳性组vs.实验组	-0.106 7	-0.312 0~0.098 61	ns	0.390 8

药物ID	成分名称	OB值/%	DL值
MOL005107	乙氧基白屈菜红素(Ethoxychelerythrine)	62.21	0.88
MOL001458	黄连碱(Coptisine)	30.67	0.86
MOL001454	小檗碱(Berberine)	36.86	0.78
MOL001987	β-谷甾醇(β-Sitosterol)	33.94	0.70
MOL000785	黄藤素(Palmatine)	64.60	0.65
MOL004763	异博尔定(Isoboldine)	39.53	0.37
MOL004093	硫唑胺(Azaleatin)	54.28	0.30
MOL000098	槲皮素(Quercetin)	46.43	0.28
MOL008099	鸦胆子苷M(Yadanzioside M)	45.04	0.23
MOL001495	亚麻酸乙酯(Ethyl linolenate)	46.10	0.20

蛋白名称	成分名称	LibDock得分
SRC	小檗碱(Berberine)	105.047
MAPK1	不列颠宁(Britanin)	105.331
MAPK14	不列颠宁(Britanin)	112.545
MAPK14	小檗碱(Berberine)	107.844
JAK1	不列颠宁(Britanin)	101.951
EGFR	弗拉辛(Flazin)	111.153
JAK3	异博尔定(Isoboldine)	133.000
IL2	硫唑胺(Azaleatin)	120.744
IL2	异鼠李素(Isorhamnetin)	119.912
JUN	鸦胆子苷H(Yadanzioside H)	138.656
JUN	鸦胆子苷J(Yadanzioside J)	116.164