Revealing the therapeutic mechanism of Hedyotis diffusa-Sculellaria barbata in lung cancer through network pharmacology and molecular docking

doi:10.21203/rs.3.rs-3785942/v1

Objective: Elucidating the therapeutic mechanism of the combination of Hedyotis diffusa and Sculellaria barbata for lung cancer through network pharmacology and molecular docking.

Method: We employed network pharmacology and molecular docking techniques to identify effective components and targets using the TCMSP database. Furthermore, relevant lung cancer targets were retrieved from the GEO database. The main targets of Hedyotis diffusa- Sculellaria barbata for lung cancer treatment were visualized using Cytoscape. Cytoscape 3.7.1 and the STRING database platform were utilized to respectively construct the active ingredient-target network map and the protein-protein interaction (PPI) network of common target proteins. To identify the therapeutic targets of the combination of Hedyotis diffusaand Sculellaria barbata for lung cancer treatment, a topological analysis was performed using cytoNCA, followed by network integration. The molecular targets and genes obtained were subjected to gene ontology (GO) function enrichment analysis and KEGG pathway analysis. The molecular docking of key action targets and major active ingredients was conducted using the AutoDockTools 1.5.6 software.

Results: A total of 32 bioactive constituents were identified in the medicinal combination of Hedyotis diffusa-Sculellaria barbata, comprising 7 compounds from Hedyotis diffusa and 25 compounds from Sculellaria barbata. Notably, quercetin, baicalin, flavonoids, and luteolin emerged as key components with significant relevance to lung cancer treatment. The target analysis revealed AKT1, HSP90AA1, SRC, CASP3, and MAPK1 as potential therapeutic targets for lung cancer treatment. The GO enrichment analysis revealed the involvement of 376 biological process-related items, 50 cell composition-related items, and 116 molecular function-related items in the targets. A total of 51 pathways were identified through KEGG pathway enrichment analysis. The molecular docking results confirmed the stable binding of ATK1 to the active ingredients baicalein and luteolin, HSP90AA1 to baicalein and flavonoids, SRC to all four active ingredients, and CASP3 to quicaletin and luteolin.

Conclusion: The treatment of lung cancer with Hedyotis diffusa-Sculellaria barbata exhibits the characteristics of being multi-component, multi-target, and multi-pathway, thereby providing a scientific foundation for both basic research and clinical application.

Hedyotis diffusa

Sculellaria barbata

Lung carcinoma

Network pharmacology analysis of traditional Chinese medicine

Molecular docking studies

Lung cancer is a malignant neoplasm primarily originating from the bronchial mucosal epithelium. According to global cancer statistics, it is estimated that over 2 million new cases of lung cancer are diagnosed annually, with Asia exhibiting the highest incidence rates^1–6. In recent years, there has been a gradual increase in the incidence of lung cancer in China. It is estimated that in 2020, China will witness approximately 820,000 new cases of lung cancer and experience around 715,000 deaths related to this disease^3,4. There are two distinct subtypes of lung cancer, namely non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC). NSCLC accounts for the majority of cases and is characterized by high malignancy, rapid cellular proliferation, and propensity for distant metastasis.

Hedyotis diffusa, an annual herbaceous plant widely distributed in eastern Asia including China and Japan⁷, belongs to the Hedyotis genus within the Rubiaceae family. The medicinal plant Hedyotis diffusa possesses therapeutic properties such as heat-clearing and detoxification, pain relief and diffusion, diuresis and dehumidification, among others. It is commonly employed in the treatment of conditions including pulmonary fever, asthma-induced cough, gastroenteritis, appendicitis, urinary tract infections, pharyngitis, intestinal abscesses, dysentery and other ailments^8–11. Recent research has demonstrated its remarkable inhibitory effect on tumor proliferation¹².

Sculellaria barbata, a perennial herb belonging to the genus Scutellaria in the Lamiaceae family, is widely distributed in northeast Asia and possesses heat-clearing and detoxifying properties¹³. The extract derived from S. melabelia exhibits significant antioxidant and anti-inflammatory activities along with other important biological effects^14–16. This extract finds extensive application in treating inflammation and associated discomfort, often combined with Hedyotis diffusa to enhance efficacy and alleviate symptoms^17–19. Notably, studies have demonstrated that S. bushelia exerts diverse pharmacological effects on tumors including inhibition of cell proliferation, migration, invasion, angiogenesis suppression, as well as promotion of apoptosis^20–22.

The field of Chinese medicine network pharmacology emerged from the initial proposal of Pharmacoinformatics by Hopkins, which was later integrated with Chinese medicine. The utilization of the TCMSP database has gradually introduced the concept of TCM network pharmacology²³. By means of predicting and analyzing TCM components, action targets, and signaling pathways, it becomes possible to unveil both the active components and their mechanisms in Traditional Chinese Medicine (TCM). Molecular Docking is a computational chemistry technique employed to investigate the binding and interaction mechanisms between proteins and small molecules, such as drug candidates²⁴. This approach utilizes computer simulations to predict precise binding sites and patterns, facilitating the identification of potential drug targets, evaluation of drug candidate compounds' binding efficiency, and prediction of intermolecular affinity. By integrating network pharmacology with molecular docking, Hedyotis diffusa-Sculellaria barbata can effectively screen potential drug molecules, thereby expediting the drug development process.

1.1 Screening of the bioactive constituents and molecular targets of Hedyotis diffusa-Sculellaria barbata

The keywords Hedyotis diffusa and Sculellaria barbata were queried in the Systematic Pharmacology Database of Chinese Medicine (TCMSP). Screening conditions included "oral bioavailability (OB) ≥30%" and "drug similarity (DL) ≥0.18". By applying these criteria, we obtained information on the effective components and target profiles of Hedyotis diffusa and Sculellaria barbata drug pairs.

1.2 Acquisition of lung cancer differential genes

The keywords "lung cancer" were selected from the GEO database based on the following criteria: (1) a total sample size greater than 50; and (2) inclusion of both tumor tissue and normal tissue samples for screening. The GEO data annotations were transformed into corresponding genes, followed by differential analysis using Perl language. Finally, differential heat maps and volcano plots were generated from the GEO database.

1.3 Construction of protein-protein interaction (PPI) network for the shared target between Hedyotis diffusa-Sculellaria barbata and lung cancer.

The screening conditions for the STRING database were as follows: a total score >0.4 and "Homo sapiens" were selected, single proteins without interactions were excluded, and default settings were applied for the remaining parameters. The shared target protein-protein interaction (PPI) network was visualized using Cytoscape 3.7.1 software, and core targets were subsequently analyzed and filtered.

1.4 Gene enrichment analysis of common target of Hedyotis diffusa-Sculellaria barbata and lung cancer

The intersection between the regulatory gene of Hedyotis diffusa-Sculellaria barbata and the differential gene associated with lung cancer was analyzed to identify the target gene for this drug's activity against lung cancer. Subsequently, an active component-target network diagram of Hedyotis diffusa-Sculellaria barbata was constructed using Cytoscape 3.8.2 software.

The target genes obtained after network merging were subjected to gene ontology (GO) enrichment analysis and KEGG biological pathway enrichment analysis using CytoNCA and R language analysis tools. This resulted in the identification of biological processes (BP), molecular functions (MF), and cellular components (CC). GO enrichment was visualized through bar charts and bubble charts for MF and CC, while KEGG biological pathways were represented by bar charts, bubble charts, and a constructed KEGG network relationship in Cytoscape.

1.5 Molecular Docking

The mol2 file containing the active ingredients of drug Hedyotis diffusa-Sculellaria barbata was obtained from the tcmsp database, while the 3D structure of the core target was acquired from the RSCB PDB database. Subsequently, autodock software was employed to perform molecular docking analysis. Finally, pymol software was utilized for visualization purposes. Binding energy serves as a reliable indicator to assess the strength of interaction between a compound and its target protein. In this study, a binding energy value lower than -5 kj/mol indicates a favorable binding affinity between the target substance and its corresponding component.

2.1 Hedyotis diffusa-Sculellaria barbata main active ingredients and target prediction

A total of 131 active ingredients were retrieved from the TCMSP database. Following rigorous screening and weight adjustment, we identified 7 principal active constituents, corresponding to a total of 555 targets. Additionally, Melea officinalis exhibited 25 key bioactive components that targeted a comprehensive set of 1228 proteins (Table 1).

Table 1 Chemical constituents of the herbal pair of Hedyotis chinensis

MOL ID	Chemical composition	OB/％	DL
MOL001646	2,3-dimethoxy-6-methyanthraquinone	34.86	0.26
MOL001659	Poriferasterol	43.83	0.76
MOL001663	(4aS,6aR,6aS,6bR,8aR,10R,12aR,14bS)-10-hydroxy-2,2,6a,6b,9,9,12a-heptamethyl-1,3,4,5,6,6a,7,8,8a,10,11,12,13,14b-tetradecahydropicene-4a-carboxylic acid	32.03	0.76
MOL001670	2-methoxy-3-methyl-9,10-anthraquinone	37.83	0.21
MOL000449	Stigmasterol	43.83	0.76
MOL000358	beta-sitosterol	36.91	0.75
MOL000098	quercetin	46.43	0.28
MOL001040	(2R)-5,7-dihydroxy-2-(4-hydroxyphenyl)chroman-4-one	42.36	0.21
MOL012245	5,7,4'-trihydroxy-6-methoxyflavanone	36.63	0.27
MOL012246	5,7,4'-trihydroxy-8-methoxyflavanone	74.24	0.26
MOL012248	5-hydroxy-7,8-dimethoxy-2-(4-methoxyphenyl)chromone	65.82	0.33
MOL012250	7-hydroxy-5,8-dimethoxy-2-phenyl-chromone	43.72	0.25
MOL012251	Chrysin-5-methylether	37.27	0.2
MOL012252	9,19-cyclolanost-24-en-3-ol	38.69	0.78
MOL002776	Baicalin	40.12	0.75
MOL012254	campesterol	37.58	0.71
MOL000953	CLR	37.87	0.68
MOL000358	beta-sitosterol	36.91	0.75
MOL012266	rivularin	37.94	0.37
MOL001973	Sitosteryl acetate	40.39	0.85
MOL012269	Stigmasta-5,22-dien-3-ol-acetate	46.44	0.86
MOL012270	Stigmastan-3,5,22-triene	45.03	0.71
MOL000449	Stigmasterol	43.83	0.76
MOL000173	wogonin	30.68	0.23
MOL001735	Dinatin	30.97	0.27
MOL001755	24-Ethylcholest-4-en-3-one	36.08	0.76
MOL002714	baicalein	33.52	0.21
MOL002719	6-Hydroxynaringenin	33.23	0.24
MOL002915	Salvigenin	49.07	0.33
MOL000351	Rhamnazin	47.14	0.34
MOL000359	sitosterol	36.91	0.75
MOL005190	eriodictyol	71.79	0.24
MOL005869	daucostero_qt	36.91	0.75
MOL000006	luteolin	36.16	0.25
MOL008206	Moslosooflavone	44.09	0.25
MOL000098	quercetin	46.43	0.28

2.2 Target identification in the context of lung cancer

In the GEO database, we identified data GSE116959 that met our criteria with a sample size exceeding 50 and inclusion of both tumor tissue and normal tissue samples as screening conditions. From this dataset, we obtained a total of 68 samples comprising 57 lung cancer samples and 11 normal control group samples. Subsequently, microarray analysis was conducted on these 68 samples to identify differentially expressed gene sequences associated with the disease progression and determine their biological significance. After statistical analysis with a significance level of P < 0.05, we identified a total of 401 differentially expressed genes, comprising 130 up-regulated genes and 271 down-regulated genes. Subsequently, differential heat maps and volcano plots were generated to visually represent the gene expression patterns (Figure 2A and Figure 2D). In these visualizations, up-regulated genes are depicted in red while slightly down-regulated genes are represented in green.

2.3 Analysis of PPI protein interaction network

The intersection targets were imported into the STRING database to retrieve protein-protein interaction (PPI) network information, and the resulting PPI network of target proteins was visualized using Cytoscape 3.7.1 software (Figure 2B-C). The graph consists of 19 nodes and 49 edges. In terms of Betweenness centrality, the top five core targets in the PPI network are AKT1, HSP90AA1, SRC, CASP3, and MAPK1 (Table 2).

Table 2 PPI core network targets of Hedyotongue-Schellaria

Serial number	Gene name	Betweenness
1	AKT1	1957.65427
2	HSP90AA1	1100.419189
3	SRC	1046.809629
4	CASP3	887.9259625
5	MAPK1	723.4805155

2.4 Enrichment analysis of shared target genes between Hedyotis diffusa-Sculellaria barbata and lung cancer.

By intersecting the genes associated with effective drug targets of Hedyotis diffusa-Sculellaria barbata with the differentially expressed genes in lung cancer, a total of 18 potential therapeutic targets against lung cancer were identified. The corresponding drugs and their target genes in lung cancer were comprehensively analyzed (Figure 2E). In this figure, green represents the components derived from Hedyotis diffusa, purple represents those from Sculellaria barbata, and blue denotes the lung cancer-associated genes. Notably, there is an overlap (depicted in red) between these two plant species. Among them, quercetin, wogonin, Chrysin-5-methylether, and luteolin exhibit multi-targeted activities, indicating their potential as crucial components in the therapeutic management of lung cancer. The results also demonstrated the efficacy of the drug combination Hedyotis diffusa-Sculellaria barbata in treating lung cancer through its multi-component and multi-target interactions.

The R language was employed to conduct gene ontology (GO) enrichment analysis on the acquired TCM compound network, yielding the corresponding BP and CC histograms as well as a bubble diagram (Figure 3A-C). The X-axis represents the count, with taller columns indicating greater numbers of genes in each category and more significant enrichments. The Y-axis denotes enrichment items, where a redder color signifies higher significance of enrichment, accompanied by larger bubbles in the bubble diagram. By conducting GO function analysis, a total of 376 biological process terms were identified, primarily associated with the positive regulation of kinase activity, transmembrane receptor protein tyrosine kinase signaling pathway, and positive regulation of cell proliferation. Among these terms, 50 cellular component terms were predominantly enriched in receptor complex, macromolecular complex, plasma membrane, etc. Additionally, 116 molecular function terms were mainly concentrated in transmembrane receptor protein tyrosine kinase activity, protein tyrosine kinase activity, ATP binding and other related functions.

The KEGG analysis was performed using CytoNCA and R language, resulting in the identification of a total of 51 pathways. Figure 4A-B presents column graphs and bubble graphs depicting the enrichment analysis results for the first 18 pathways (Table 3). Additionally, Figure 4C presents a network diagram illustrating the findings of KEGG enrichment analysis. The increase in the size of cones corresponds to a higher number of enriched genes within the pathway, while larger rectangles indicate an increased involvement of genes in the pathway. The findings indicate that Hedyotis diffusa-Sculellaria barbata may have therapeutic potential for lung cancer by modulating various pathways, including those involved in fluid shear stress and diseases such as atherosclerosis, rheumatoid arthritis, chemical carcinogenic receptor activation, cell aging, pertussis signaling pathways.

Table 3 KEGG enrichment analysis pathway

ID	Description	geneID	Count
hsa05418	Fluid shear stress and atherosclerosis	FOS/CAV1/IL1B/IL1A/KDR	5
hsa05323	Rheumatoid arthritis	FOS/MMP1/IL1B/IL1A	4
hsa05207	Chemical carcinogenesis - receptor activation	ADRB2/AR/FOS/BIRC5/E2F1	5
hsa04218	Cellular senescence	CDK1/IL1A/E2F1/IGFBP3	4
hsa05133	Pertussis	FOS/IL1B/IL1A	3
hsa05140	Leishmaniasis	FOS/IL1B/IL1A	3
hsa04657	IL-17 signaling pathway	FOS/MMP1/IL1B	3
hsa05215	Prostate cancer	PLAU/AR/E2F1	3
hsa04933	AGE-RAGE signaling pathway in diabetic complications	IL1B/IL1A/COL3A1	3
hsa04926	Relaxin signaling pathway	FOS/MMP1/COL3A1	3
hsa04380	Osteoclast differentiation	FOS/IL1B/IL1A	3
hsa05162	Measles	FOS/IL1B/IL1A	3
hsa04010	MAPK signaling pathway	FOS/IL1B/IL1A/KDR	4
hsa05219	Bladder cancer	MMP1/E2F1	2
hsa05332	Graft-versus-host disease	IL1B/IL1A	2
hsa04940	Type I diabetes mellitus	IL1B/IL1A	2
hsa04932	Non-alcoholic fatty liver disease	FOS/IL1B/IL1A	3

2.5 Molecular docking verification

The betweenness centrality measure was employed for screening Hedyotis diffusa-Sculellaria barbata, resulting in the identification of AKT1, HSP90AA1, SRC, CASP3, and MAPK1 as the top 5 core drug targets. Based on target density ranking from high to low, four core components including quercetin, wogonin, Chrysin-5-methylether, and luteolin were obtained. PyMOL software and AutoDockTools-1.5.6 software were utilized to establish connections between these core components and their respective targets, with partial docking results visualized using PyMOL software (Table 4 and Figure5).

Table 4 Values of binding energy for docking

Compound	Target
Compound	ATK1	HSP90AA1	SRC	CASP3	MAPK1
quercetin	×	-5.02	-6.00	-5.25	×
wogonin	-5.18	-6.25	-5.59	×	×
Chrysin-5-methylether	-5.37	-6.05	-5.59	×	×
luteolin	-5.46	×	-6.02	-5.07	×

Lung cancer is a highly lethal malignancy and one of the most prevalent cancers globally^3,4, arising from aberrantly proliferating and dividing cells in lung tissue. It typically originates in bronchial epithelial or alveolar cells and has the potential to metastasize to distant sites. The pathogenesis of lung cancer involves genetic mutations, cellular proliferation, invasion, and metastasis. In its early stages, lung cancer often remains asymptomatic; however, as the tumor progresses, common manifestations include persistent coughing, hemoptysis, chest pain, dyspnea, hoarseness of voice, recurrent respiratory infections, unintentional weight loss, and fatigue.

Traditional Chinese medicine has demonstrated advantages in the treatment of lung cancer, including comprehensive regulation, mitigation of toxic side effects, enhancement of immune response, and personalized treatment; however, further investigation is required to elucidate its underlying mechanisms. The intricate interplay among multiple components in TCM compound preparations may exert influence on tumor growth, metastasis, and the immune system. The herb is believed to possess significant therapeutic properties in TCM, including heat-clearing and detoxifying effects, promotion of blood circulation and removal of blood stasis, reduction of swelling, pain relief in tumor treatment, as well as effective inhibition of tumor growth, improvement of physical condition, and enhancement of immunity. The plant contains numerous active components, including alkaloids, flavonoids, sesquiterpenoids, and polysaccharides. Extensive research has demonstrated their anti-tumor, antioxidant, anti-inflammatory, and immunomodulatory effects, suggesting potential therapeutic applications in cancer treatment. However, the precise identification of active compounds and underlying mechanisms remains elusive; thus further investigations are warranted to elucidate their specific therapeutic mechanisms and ensure safety.

In the clinical treatment of TCM, the combination of Hedyotis diffusa and Sculellaria barbata is frequently employed, as it is believed to exhibit a synergistic effect that enhances its anti-tumor efficacy. Both Hedyotis diffusa and Sculellaria barbata contain similar bioactive constituents such as alkaloids and flavonoids, which contribute to their heat-clearing and detoxifying properties, promotion of blood circulation, removal of blood stasis, regulation of immune function, among others. This combined therapeutic approach thus facilitates improved treatment outcomes.

Studies have demonstrated the anti-tumor properties of Sculellaria barbata in various cancer types, including lung, breast, and pancreatic cancers, by exerting inhibitory effects on cancer cell growth and inducing apoptosis^12,25–27. Furthermore, it has been reported that polysaccharides present in Sculellaria barbata can upregulate the expression level of miR-195-5p, thereby suppressing the invasion and migration of lung cancer cells. These findings suggest the potential anticancer efficacy of machiea officinalis²⁸.

The network pharmacological prediction of anti-lung cancer for Hedyotis diffusa and Sculellaria barbata was conducted in this study and validated using molecular docking technology. The TCMSP database was utilized to obtain the OB, DL, and relevant target data of Hedyotis diffusa and Sculellaria barbata. By matching components with targets, performing pathway enrichment analysis, and ranking by degree, it was discovered that Hedyotis diffusa and Sculellaria barbata contains quercetin, wogonin, Chrysin-5-methylether, and luteolin as its constituents which correspond to multiple targets. Quercetin, a polyphenolic flavonoid compound, has attracted much attention in the field of cancer research^29,30. In related studies, quercetin has been found to induce pro-apoptotic autophagy in non-small cell lung cancer cell lines through SIRT1/AMPK signaling pathway in vitro. In addition, quercetin can also inhibit the growth and promote apoptosis of A549 and H1299 lung cancer cells^31–33. Baicalein has been proved to have certain potential in anti-tumor, it can affect the proliferation, metastasis and invasion of a variety of cancer cells, and can also induce apoptosis of cancer cells. Baicalin influences the growth and metastasis of tumor cells by mediating and regulating the c-Myc/Skp2/Fbw7α and HDAC1/HDAC2 pathways^34,35. Flavonoids have significant anti-tumor effects, which can affect the proliferation of lung cancer by regulating PI3K/Akt/JAK-STAT3 and other pathways. At the same time, flavonoids can also affect cell apoptosis and autophagy^32,36. Luteolin is also a kind of flavonoid, which has anti-inflammatory and anti-allergic effects³⁷. Antitumor effects have been demonstrated in some lung cancer studies, and in lung cancer cells, luteolin acts as a radiosensitizer to increase apoptotic cell death by activating the p38/ROS/caspase cascade^38,39.

In this study, we identified five core target genes, namely ATK1, HSP90AA1, SRC, CASP3 and MAPK1. Serine/threonine protein kinase Akt (also known as protein kinase B or PKB) is a pivotal regulator of cellular signaling pathways. Akt kinase plays a crucial role in regulating cell survival, proliferation and metabolism. Notably, the ATK1 pathway exhibits robust activation in lung cancer tissues⁴⁰. The gene HSP90AA1 encodes Heat Shock Protein 90 Alpha, which plays a pivotal role in cellular stress response, heat shock reaction, and intracellular signaling. This protein is crucial for various cellular processes, particularly during the adaptive response to environmental changes and stressors. Notably, HSP90AA1 exhibits significantly elevated transcriptional activity and expression levels in lung cancer patient tissues⁴⁰. SRC, a member of the tyrosine kinase family, is a key regulator in cell signaling and plays crucial roles in various cellular processes including proliferation, differentiation, migration, adhesion, and apoptosis. Moreover, SRC has emerged as a significant oncoprotein implicated in cancer progression, invasion, metastasis, and drug resistance across multiple malignancies such as colorectal, breast pancreatic gastric and pulmonary cancers⁴¹. CASP3, also referred to as Caspase 3, is a member of the cysteine protease family and functions as a crucial protein in the apoptosis pathway by promoting programmed cell death. Its expression can be observed in both the nucleus and cytoplasm of lung cancer cells, which correlates with cancer development and other pathological conditions⁴². MAPK1, a member of the serine/threonine protein kinase family and the MAP kinase family, plays a pivotal role in intracellular signaling pathways and governs crucial biological processes such as cell growth, proliferation, and differentiation. Moreover, it exerts regulatory control over tumor proliferation and metastasis, thereby modulating tumorigenesis^43,44.

The GO results demonstrated significant enrichment of relevant target genes in diverse biological processes, cellular components, and molecular functions. These included positive regulation of kinase activity, transmembrane receptor protein tyrosine kinase signaling pathways, and protein tyrosine kinase activity. KEGG analysis using CytoNCA and R language identified a total of 51 pathways, with the initial 18 pathways exhibiting enrichment for analysis. Hedyotis diffusa and Sculellaria barbata have potential implications in lung cancer treatment by targeting fluid shear stress as well as atherosclerosis, rheumatoid arthritis, chemical carcinogenic receptor activation, cell aging, pertussis signaling pathway among other pathways. Molecular docking results revealed stable binding between ATK1 and the active ingredients baicalein and luteolin; HSP90AA1 and the active ingredients baicalein and flavonoids; SRC with four active ingredients; CASP3 with the active ingredients quercetin and luteolin. In conclusion, this study suggests that ATK1, HSP90AA1, SRC, and CASP3 could serve as potential targets for lung cancer treatment with SRC being the most promising target.

The present study systematically investigated the potential role of " Hedyotis diffusa and Sculellaria barbata " in the treatment of lung cancer using network pharmacology and molecular docking technology based on traditional Chinese medicine. By analyzing its main chemical constituents, targeted genes, and signaling pathways, it was revealed that this herbal remedy possesses therapeutic properties characterized by multi-component, multi-target, and multi-pathway effects. This study not only offers a novel perspective for lung cancer treatment but also establishes a robust theoretical foundation for future systematic research. These findings are anticipated to provide crucial guidance for developing more effective strategies with reduced side effects in the management of lung cancer while deepening our understanding of the mechanisms underlying traditional Chinese medicine (TCM) treatments and exploring their clinical potential.

Hedyotis diffusa-Sculellaria barbata contains 32 active ingredients, among which Quercetin 、wogonin、Chrysin-5-methylether and luteolin are closely associated with lung cancer treatment. At the same time, AKT1, HSP90AA1, SRC, CASP3 and MAPK1 are potential core targets for the treatment of lung cancer.Results from target identification and pathway enrichment analysis provide valuable insights into the molecular mechanisms of Hedyotis diffusa-Sculellaria barbata in treating lung cancer, highlighting potential targets, pathways, and involved biological processes.

Author Contribution

Zhu Dantong completed the manuscript writing of network pharmacology and molecular docking, Liang Shiying completed the related data collation, Li Shun and Ren Xijing produced Figures 1 to 6, Chen Jianjun and Jiang Mingzhe were responsible for the production of related tables, and Bu Fanshu completed the manuscript review.

Acknowledgments

This work was supported by a grant from the Shenyang science and technology plan fund project (213463) and Liaoning Province science and technology planning project (2021JH2/10300091).

Declaration of Interest statement

The authors declare that they have no conflicts of interest with the contents of this article.

Disclosure Statement

There is no dispute of interest among the authors listed in this manuscript.

Zhang S et al (2021) Cancer incidence and mortality in China, 2015. J Natl Cancer Cent 1:2–11
Siegel RL, Miller KD, Fuchs HE, Jemal A (2022) Cancer statistics, 2022. CA. Cancer J Clin 72:7–33
Sung H et al (2021) Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA. Cancer J Clin 71:209–249
Cao W, Chen H-D, Yu Y-W, Li N, Chen W-Q (2021) Changing profiles of cancer burden worldwide and in China: a secondary analysis of the global cancer statistics 2020. Chin Med J (Engl) 134:783–791
Luo Y-H et al (2021) Lung Cancer in Republic of China. J Thorac Oncol Off Publ Int Assoc Study Lung Cancer 16:519–527
Gao S et al (2020) Lung Cancer in People’s Republic of China. J Thorac Oncol Off Publ Int Assoc Study Lung Cancer 15:1567–1576
Li M et al (2013) Application of novel loop-mediated isothermal amplification (LAMP) for rapid authentication of the herbal tea ingredient Hedyotis diffusa Willd. Food Chem 141:2522–2525
Chen R, He J, Tong X, Tang L, Liu M (2016) The Hedyotis diffusa Willd. (Rubiaceae): A Review on Phytochemistry, Pharmacology, Quality Control and Pharmacokinetics. Mol Basel Switz 21:710
Lin J et al (2013) Hedyotis diffusa Willd extract suppresses Sonic hedgehog signaling leading to the inhibition of colorectal cancer angiogenesis. Int J Oncol 42:651–656
Wei L et al (2012) Scutellaria barbata D. Don inhibits tumor angiogenesis via suppression of Hedgehog pathway in a mouse model of colorectal cancer. Int J Mol Sci 13:9419–9430
Lin C-C et al (2011) Extract of Hedyotis diffusa Willd influences murine leukemia WEHI-3 cells in vivo as well as promoting T- and B-cell proliferation in leukemic mice. Vivo Athens Greece 25:633–640
Lee H-Z et al (2011) Clarification of the phenotypic characteristics and anti-tumor activity of Hedyotis diffusa. Am J Chin Med 39:201–213
Wang L et al (2020) A review of the ethnopharmacology, phytochemistry, pharmacology, and quality control of Scutellaria barbata D. Don. J Ethnopharmacol 254:112260
Ye C-L, Huang Q (2012) Extraction of polysaccharides from herbal Scutellaria barbata D. Don (Ban-Zhi-Lian) and their antioxidant activity. Carbohydr Polym 89:1131–1137
Wang Z et al (2016) Scutellarin protects cardiomyocyte ischemia-reperfusion injury by reducing apoptosis and oxidative stress. Life Sci 157:200–207
Liu H-L, Kao T-H, Shiau C-Y, Chen B-H (2018) Functional components in Scutellaria barbata D. Don with anti-inflammatory activity on RAW 264.7 cells. J Food Drug Anal 26:31–40
Liu H, Xue J-X, Li X, Ao R, Lu Y (2013) Quercetin liposomes protect against radiation-induced pulmonary injury in a murine model. Oncol Lett 6:453–459
Qin M et al (2017) Protective efficacy of inhaled quercetin for radiation pneumonitis. Exp Ther Med 14:5773–5778
Jang H et al (2019) Baicalein Mitigates Radiation-Induced Enteritis by Improving Endothelial Dysfunction. Front Pharmacol 10:892
Guo F, Yang F, Zhu Y-H (2019) Scutellarein from Scutellaria barbata induces apoptosis of human colon cancer HCT116 cells through the ROS-mediated mitochondria-dependent pathway. Nat Prod Res 33:2372–2375
Zheng X, Kang W, Liu H, Guo S (2018) Inhibition effects of total flavonoids from Sculellaria barbata D. Don on human breast carcinoma bone metastasis via downregulating PTHrP pathway. Int J Mol Med 41:3137–3146
Liu J et al (2018) A Novel Systems Pharmacology Method to Investigate Molecular Mechanisms of Scutellaria barbata D. Don for Non-small Cell Lung Cancer. Front Pharmacol 9:1473
Jiashuo WU, Fangqing Z, Zhuangzhuang LI, Weiyi J, Yue S (2022) Integration strategy of network pharmacology in Traditional Chinese Medicine: a narrative review. J Tradit Chin Med Chung Tsa Chih Ying Wen Pan 42:479–486
Pinzi L, Rastelli G (2019) Molecular Docking: Shifting Paradigms in Drug Discovery. Int J Mol Sci 20:4331
Wang L et al (2019) Green synthesis of gold nanoparticles from Scutellaria barbata and its anticancer activity in pancreatic cancer cell (PANC-1). Artif Cells Nanomedicine Biotechnol 47:1617–1627
Yang Y et al (2020) Ethyl Acetate Fraction from Hedyotis diffusa plus Scutellaria barbata Exerts Anti-Breast Cancer Effect via miR-200c-PDE7B/PD-L1-AKT/MAPK Axis. Evid.-Based Complement. Altern. Med. ECAM. 3587095 (2020)
Lv Y-X, Pan H-R, Song X-Y, Chang Q-Q, Zhang D-D (2021) Hedyotis diffusa plus Scutellaria barbata Suppress the Growth of Non-Small-Cell Lung Cancer via NLRP3/NF- κ B/MAPK Signaling Pathways. Evid. Based Complement. Alternat. Med. 1–8 (2021)
Xue S, Geng A, Lian T, Liu Y (2022) Scutellaria barbata D. Don inhibits cervical cancer cell proliferation, migration, and invasion via miR-195-5p/LOXL2 axis. Toxicol Res 11:804–811
Jeong J-H, An JY, Kwon YT, Rhee JG, Lee YJ (2009) Effects of low dose quercetin: cancer cell-specific inhibition of cell cycle progression. J Cell Biochem 106:73–82
Deng S et al (2019) Targeting autophagy using natural compounds for cancer prevention and therapy. Cancer 125:1228–1246
Nguyen TTT et al (2004) The role of activated MEK-ERK pathway in quercetin-induced growth inhibition and apoptosis in A549 lung cancer cells. Carcinogenesis 25:647–659
Guo H et al (2021) Quercetin induces pro-apoptotic autophagy via SIRT1 / AMPK signaling pathway in human lung cancer cell lines A549 and H1299 in vitro. Thorac Cancer 12:1415–1422
Zheng S-Y, Li Y, Jiang D, Zhao J, Ge J-F (2012) Anticancer effect and apoptosis induction by quercetin in the human lung cancer cell line A-549. Mol Med Rep 5:822–826
Chen X et al (2013) Wogonin Has Multiple Anti-Cancer Effects by Regulating c-Myc/SKP2/Fbw7α and HDAC1/HDAC2 Pathways and Inducing Apoptosis in Human Lung Adenocarcinoma Cell Line A549. PLoS ONE 8:e79201
Cao H-J et al (2022) A Mixture of Baicalein, Wogonin, and Oroxylin-A Inhibits EMT in the A549 Cell Line via the PI3K/AKT-TWIST1-Glycolysis Pathway. Front Pharmacol 12:821485
Zanoaga O et al (2019) Progress in Research on the Role of Flavonoids in Lung Cancer. Int J Mol Sci 20:4291
Nabavi SF et al (2015) Luteolin as an anti-inflammatory and neuroprotective agent: A brief review. Brain Res Bull 119:1–11
Ma L et al (2015) Luteolin exerts an anticancer effect on NCI-H460 human non-small cell lung cancer cells through the induction of Sirt1-mediated apoptosis. Mol Med Rep 12:4196–4202
Luteolin acts as a radiosensitizer in non–small cell lung cancer cells by enhancing apoptotic cell death through activation of a p38/ROS/caspase cascade - PubMed. https://pubmed.ncbi.nlm.nih.gov/25586525/
Niu M et al (2022) Targeting HSP90 Inhibits Proliferation and Induces Apoptosis Through AKT1/ERK Pathway in Lung Cancer. Front Pharmacol 12:724192
Hsu P-C, Yang C-T, Jablons DM, You L (2020) The Crosstalk between Src and Hippo/YAP Signaling Pathways in Non-Small Cell Lung Cancer (NSCLC). Cancers 12:1361
Takata T et al (2001) Clinical significance of caspase-3 expression in pathologic-stage I, nonsmall-cell lung cancer. Int J Cancer 96 Suppl, 54–60
Guégan J-P, Ezan F, Théret N, Langouët S, Baffet G (2013) MAPK signaling in cisplatin-induced death: predominant role of ERK1 over ERK2 in human hepatocellular carcinoma cells. Carcinogenesis 34:38–47
Lin L et al (2014) CXCR7 stimulates MAPK signaling to regulate hepatocellular carcinoma progression. Cell Death Dis 5:e1488

No competing interests reported.

Revealing the therapeutic mechanism of Hedyotis diffusa-Sculellaria barbata in lung cancer through network pharmacology and molecular docking

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Abstract

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Background

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Version 1