Mechanism of Coriander in the Treatment of Osteoarthritis Based on Network Pharmacology

Chunfeng WU, Li KONG, Huajiang ZUO

1.Liuzhou People’s Hospital Affiliated to Guangxi Medical University, Liuzhou 545006, China; 2.Guangxi University of Science and Technology, Liuzhou 545006, China

Abstract [Objectives]To explore the mechanism of coriander in the treatment of osteoarthritis by means of network pharmacology.[Methods]The effective active components of coriander and their targets were screened from TCMSP database.The targets related to osteoarthritis were screened from CTD database.The targets of active components were matched with the targets of diseases to obtain potential targets.PPI was constructed by using STRING database, and key target proteins were selected according to the degree value and the medium number.The DAVID database is used to analyze the biological functions and pathways of potential targets.[Results]15 active components of coriander were screened out by using OB and DL, and corresponding 102 targets were found, of which 45 were intersected with osteoarthritis-related targets, and PPI network contains 45 targets.The enrichment analysis of go includes 100 items, including 72 items related to biological processes, 17 items related to cell components, 11 items related to molecular functions, and 62 items related to KEGG pathway.[Conclusions]The results of network pharmacological analysis in this study indicate that the effective active ingredients of coriander mainly act on key targets such as CASP3, IL6, ALB, VEGFA, EGFR, MYC, CCND1, PPARG, FOS, RELA, etc. Multiple signal pathways such as p53 and PIK-AKt regulate the expression of genes to achieve the purpose of treating osteoarthritis, and lay a certain foundation for the mechanism of coriander to treat osteoarthritis.

Key words Coriander, Osteoarthritis, Network pharmacology, Target, Signal pathway

1 Introduction

Osteoarthritis(OA)is a chronic disease with high incidence.Its main clinical manifestations are joint swelling, pain, deformity and dyskinesia.It can lead to disability in severe cases.At present, the prevalence rate of people over the age of 60 reaches 50% and the prevalence rate of people over the age of 75 is as high as 80%[1-2].Studies have shown that the risk of disability caused by osteoarthritis is 40% and 47% in men and women respectively, and the proportion is larger in obese people[3].With the intensification of the aging of the population in China, the incidence of osteoarthritis is gradually increasing[4].From the perspective of traditional Chinese medicine, osteoarthritis falls into the category of "bone obstruction disease"[5-6].Traditional Chinese medicine has the advantages of good curative effect, few adverse reactions and rich choice in related treatment[7].As a kind of edible vegetable, coriander has the functions of anti-oxidation[8-9], anti-inflammation[9], dispelling wind and treating rheumatism[10], and can be effectively used in the treatment of osteoarthritis[9].However, the effective components of coriander are complex, including volatile oils and flavonoids, such as carotene[10], quercetin[11]and luteolin[12], all of which have the effect of treating osteoarthritis, but there is no research to clarify the mechanism of coriander in the treatment of osteoarthritis.Network pharmacology is a branch of pharmacology that uses network methods to analyze the synergistic relationship between drugs and diseases & targets[13].It represents a new concept and model of modern biomedicine[14], and provides a new strategy for drug research and development.Wu Danetal[15].predicted the key targets of antidepressant action of Bupleurum through protein interaction network, and verified its molecular docking.Zhang Liangetal[16].used network pharmacology to study the mechanism of anti-tumor molecules ofEchinochloapaniculata.Li Jingetal[17].adopted the method of reverse pharmacophore screening and analyzed with GooLGeN antitussive and expectorant drugs, to obtain the important targets of coltsfoot flower in relieving cough and reducing sputum, and analyzed the mechanism of coltsfoot flower in relieving cough and reducing sputum by GO enrichment analysis and KEGG enrichment analysis.

Therefore, based on the method of network pharmacology, this paper discusses the mechanism of effective active components of coriander in the treatment of osteoarthritis, in order to provide a theoretical basis for the treatment of osteoarthritis in the later stage.

2 Materials and methods

2.1 Screening of active components of coriander and collection of action targetsBy searching the traditional Chinese medicine system pharmacology database and analysis platform(TCMSP)(http://tcmspw.com/), the main chemical constituents of coriander were collected and the chemical composition database of coriander was established.On this basis, according to the oral bioavailability(OB)≥30% and drug-likeness(DL)≥0.18, the selected candidate compounds were used as the active components of coriander.The target of the above active components of coriander was searched by TCMSP database, and the prediction target was imported into UniProt database(https://www.uniprot.org/).The non-human target was removed, and the name of the prediction target was corrected to the gene name of the target, which can be used for the analysis of other work.

2.2 Collection of potential targets of coriander in the treatment of osteoarthritisThrough the CTD database(http://ctdbase.org/), using "osteoarthritis" as the keyword, the related targets of osteoarthritis were searched.According to the Inference Score sequence provided by the database, the first 500 targets were selected from high to low, and matched with the active components of coriander in Section2.1to get the intersection target, that is, the potential target of coriander in the treatment of osteoarthritis.

2.3 Construction of active component-potential target networkThe active components and potential action targets of coriander were imported into Cytoscape 3.7.0 to construct the network map of active components-action targets of coriander.

2.4 Construction of protein interaction networkThe potential interaction target was imported into STRING database(https://string-db.org/), the species was defined as human, and the minimum interaction threshold was set to 0.4.The protein interaction relationship was obtained.The results were saved in TSV format, and the node 1, node 2, Combined Score information was retained and imported into Cytoscape software to draw the network diagram of protein interaction.The network was analyzed and the analysis results were saved.The node size setting is used to reflect the size of Degree, the edge thickness is used to reflect the size of Combined Score, and the final protein interaction network diagram is obtained.This study is based on the following parameters to evaluate the importance of nodes in the network: average shortest path, betweenness centrality, closeness centrality, clustering coefficient, degree.

2.5 Biological function and pathway analysis of potential targetsThe potential target was imported into the DAVID database(https://david.ncifcrf.gov/)with Select Identifier set to OFFICIAL GENE SYMBOL, List set to Gene List.The species was defined as human, and the potential targets were analyzed by GO biological process enrichment analysis and KEGG pathway analysis.Pwas set to be less than 0.05, and Origin 7.5 was used to draw the map.

3 Results and analysis

3.1 Screening of active components of coriander and collection of action targetsThrough the search in TCMSP database, 297 coriander compounds were obtained, and 15 compounds(the active components of coriander)were selected withOB≥30%,DL≥0.18, as shown in Table 1.

Table 1 Information of active components of coriander

3.2 Collection of targets of coriander in the treatment of osteoarthritisThe targets of active components of coriander were searched in TCMSP database, among which quercetin had the most active targets(154), followed by luteolin(57), β-sitosterol(38)and β-carotene(22).A total of 386 targets were retrieved, and 102 targets were obtained after conversion by Uniprot database and removal of non-human and repeated targets.

Using "osteoarthritis" as the keyword, the related targets of osteoarthritis were searched in CTD database.According to the Inference Score sequence provided by the database, the first 500 targets were selected from high to low, and matched with the above active components of coriander to get 45 intersection targets, that is, the potential targets of coriander in the treatment of osteoarthritis, as shown in Fig.1.

Fig.1 Venny diagram of targets of active components of coriander and osteoarthritis related diseases

3.3 Construction of active component-potential target networkThe active components and potential action targets of coriander were imported into Cytoscape to construct the active component-action target network diagram, as shown in Fig.2.A total of 62 nodes(1 drug, 1 disease, 15 active components, 45 potential targets)and 144 edges are involved in the figure.In the network, the purple node represents coriander, the green node represents osteoarthritis, the red node represents the active component of coriander, and the blue node represents the potential target, and the edge represents the relationship between coriander and active components, between active components and potential targets, between osteoarthritis and potential targets.It can be seen from Fig.2 that the same active component can correspond to different targets, and the same target can also correspond to different active components, which fully reflects the characteristics of multi-components and multi-targets of coriander.

Fig.2 Network diagram of active component-potential target of coriander

3.4 Construction and analysis of protein interaction networkThe potential targets were imported into the STRING database, the species were defined as human beings, the protein interaction relationships were obtained, and the results were saved in TSV format.The TSV file was imported into Cytoscape to draw the network diagram of protein interaction, and analyze the topological properties of the network, as shown in Fig.3.

Fig.3 Protein interaction network

In the figure, the nodes represent the targets, and the edges represent the connections between the targets, involving a total of 45 nodes and 389 edges.The size of the node reflects the size of the Degree value.The larger the node is, the larger the corresponding Degree value is.The thickness of the edge represents Combine score, and the thicker the edge, the larger the Combine score.After calculation, the average connection degree was about 17.29, the average shortest path length was about 1.63, the betweenness centrality was about 0.015, and the average clustering coefficient was about 0.70, respectively.The node with large value(Degree)is considered to be the central node.In this network, there are 20 nodes whose values are higher than the average, indicating that these 20 nodes are the core nodes of the protein interaction network and may be the key targets for coriander in the treatment of osteoarthritis(Table 2).

Table 2 Network topology properties of key targets

3.5 Biological function and pathway analysis of potential targetsThe potential targets were imported into DAVID database for GO and KEGG pathway enrichment analysis, and PValue was set to be less than 0.05.Through GO enrichment analysis, it was found that the potential targets were mainly concentrated in 72 biological processes(BP), including negative regulation of apoptotic process, positive regulation of transcription from RNA polymerase II promoter, positive regulation of peptidyl-serine phosphorylation, transcription of DNA template, response to drugs and so on.There are 17 kinds of cells, including nucleus, extracellular space, cytosol, perinuclear region of cytoplasm, Golgi apparatus and so on.11 molecular functions include protein heterodimerization activity, chromatin binding, heme binding, iron ion binding, fibronectin binding and so on.The top 10 biological functions are listed according to thePvalue, as shown in Fig.4A-C.KEGG pathway enrichment analysis showed that the potential targets involved 62 signal pathways, including cancer signal pathway, hepatitis B virus, prostate cancer, colorectal cancer, TNF signal pathway, HTLV-I infection, Legionellosis, PI3K-Akt signal pathway, p53 signal pathway,etc.The first 20 paths are listed according to thePvalue, as shown in Fig.4D.The potential targets of coriander active components in the treatment of osteoarthritis involve a variety of biological processes and multiple signal pathways, indicating that coriander can play a role in the treatment of osteoarthritis by regulating these biological processes and signal pathways.

Note: A.Biological process analysis; B.Cell component analysis; C.Molecular function analysis; D.KEGG pathway enrichment analysis.

4 Discussion

In this study, the network pharmacology method was used to explore the mechanism of effective active components of coriander in the treatment of osteoarthritis.15 active components were screened by TCMSP database, including corticosterone A, imperatorin, quercetin, luteolin, β-sitosterol and so on.These active components acted on 102 targets.According to the osteoarthritis related targets in the CTD database, 45 coriander targets for the treatment of osteoarthritis were obtained by matching.The active component-action target network diagram showed the therapeutic characteristics of multi-components and multi-targets of coriander, and the protein interaction network showed the interrelationship between potential targets.Through the analysis of its network topology properties, it was found that CASP3, IL6, ALB, VEGFA, EGFR, MYC, CCND1, PPARG, FOS, RELA and so on had high degree, and they could be used as the key targets of coriander in the treatment of osteoarthritis.In the articular cavity of patients with osteoarthritis, IL1 and IL6 showed high expression.Therefore, IL1 and IL6 can be used as reference indicators to determine the disease progression of patients with osteoarthritis[18].The results of target GO analysis showed that the treatment of osteoarthritis with coriander mainly involved biological processes including negative regulation of apoptotic process, positive regulation of transcription from RNA polymerase II promoter, positive regulation of peptidyl-serine phosphorylation, transcription of DNA template, response to drugs and so on, cellular components including nucleus, extracellular space, cytosol, perinuclear region of cytoplasm, Golgi apparatus and so on, and molecular functions including protein heterodimerization activity, chromatin binding, heme binding, iron ion binding, fibronectin binding and so on.The results of pathway analysis showed that coriander mainly involved TNF, p53, PIK-AKt and other signal pathways in the treatment of osteoarthritis.P53 is a nuclear phosphorylated protein.The expression of p53 in osteoarthritis chondrocytes is higher than that in normal chondrocytes.By down-regulating the expression of p53, it can reduce the apoptosis of chondrocytes and alleviate and prevent osteoarthritis[19].PIK-AKt signal pathway can regulate protein apoptosis, and abnormal activation of PIK-AK can promote the high expression of Bcl-2 and prevent chondrocyte apoptosis[20].

To sum up, this study applied the method of network pharmacology to study the potential mechanism of coriander in the treatment of osteoarthritis, so as to explore the target and signal pathway of coriander in the treatment of osteoarthritis.and clarify the mechanism of coriander in the treatment of osteoarthritis through a variety of active components.This provides an effective theoretical basis for the follow-up application of coriander in the treatment of osteoarthritis.