阅读说明：本技术 一种基于网络药理学解析岩藻多糖抗肝损伤作用机制的方法 (Method for analyzing anti-liver injury action mechanism of fucoidin based on network pharmacology ) 是由 苏敏 黎荣 黎镜波 梁宵 于 2021-09-06 设计创作，主要内容包括：本发明公开了一种基于网络药理学解析岩藻多糖抗肝损伤作用机制的方法,首次揭示岩藻多糖抗肝损伤关键靶点和所涉及的生物学过程,具体包括：首先通过药物-疾病数据库平台分别筛选岩藻多糖作用靶点和肝损伤疾病靶点；再将岩藻多糖和肝损伤靶点映射取交集并构建蛋白互作用网络获取核心靶点；进一步利用R软件对核心靶点进行GO生物功能和KEGG通路富集分析取得可视化结果；最后构建岩藻多糖-核心靶点-GO-KEGG-肝损伤网络并解析网络相互关系。本发明通过揭示岩藻多糖抗肝损伤作用机制,为今后岩藻多糖药物开发及肝损伤治疗靶点提供参考。(The invention discloses a method for analyzing an anti-liver injury action mechanism of fucoidin based on network pharmacology, which firstly reveals an anti-liver injury key target of the fucoidin and a related biological process, and specifically comprises the following steps: firstly, respectively screening fucoidin action targets and liver injury disease targets through a drug-disease database platform; mapping the fucoidin and the liver injury target to obtain an intersection and constructing a protein interaction network to obtain a core target; further utilizing R software to perform GO biological function and KEGG channel enrichment analysis on the core target spot to obtain a visual result; and finally, constructing a fucoidin-core target spot-GO-KEGG-liver injury network and analyzing the network interrelation. The invention provides reference for fucoidin drug development and liver injury treatment targets in the future by disclosing the mechanism of the fucoidin anti-liver injury action.)

1. A method for analyzing the mechanism of action of fucoidin against liver injury based on network pharmacology is characterized in that: the method comprises the following steps:

1) fucoidin target acquisition and correction: using a drug online database to obtain fucoidan action targets, and correcting all the targets in a protein database to match with related gene symbols;

2) obtaining a liver injury target spot and identifying an intersection target spot: searching and searching for a Liver injury candidate target point in Genecard and DisGeNET databases by taking 'Liver injury' as a keyword, and then performing intersection mapping with the fucoidin action target point obtained in the step 1) to obtain a fucoidin anti-Liver injury action target point;

3) constructing a fucoidin-liver injury protein interaction network and predicting a core target: analyzing the intersection target in the step 2) by adopting a STRING database to obtain a target protein interaction relation graph and tsv data thereof, analyzing the tsv data freedom median and the maximum freedom degree by using a networkAnalyzer module in Cytoscape3.7.1 software, wherein the upper limit and the lower limit of the screening of the core target are twice of the maximum freedom degree and the freedom degree median respectively, and the target with the topological parameters contained in the upper limit and the lower limit ranges is used as the fucoidan-liver injury core target;

4) core target biological function and enrichment analysis discuss the mechanism of action of fucoidin against liver injury: a gene ontology database GO is used for analyzing the biological function of fucoidin in resisting liver injury, a Kyoto gene and genome encyclopedia database KEGG is used for revealing a fucoidin signal channel in resisting liver injury, and finally, a corresponding bubble diagram, a histogram and a Circos circle diagram are output;

5) the construction and visualization of a 'drug-target-gene ontology function-pathway-disease' network: constructed according to the results of step 4) using cytoscape 3.7.1.

2. The method for analyzing the mechanism of action of fucoidan against liver injury based on cyber pharmacology according to claim 1, wherein the method comprises the following steps: the drug online database in the step 1) is selected from Swiss Target Prediction, PharmMapper, Batman and drug bank; the protein database is selected from Uniprot; the correction is carried out to match the related gene symbols, namely, the verified humanized target is selected as a screening condition, and the name of the sorted target is converted into the name of the gene to obtain the corrected medicine target.

3. The method for analyzing the mechanism of action of fucoidan against liver injury based on cyber pharmacology according to claim 1, wherein the method comprises the following steps: the step 2) of carrying out intersection mapping with the fucoidan action target spot obtained in the step 1) is to use a letter on-line tool for correcting the fucoidan action target spot in the step 1) of finishing the liver injury target spot

http:// bioinformatics. psb. element. be/webtools/Venn/Wien diagram Venn mapping intersection processing.

4. The method for analyzing the mechanism of action of fucoidan against liver injury based on cyber pharmacology according to claim 1, wherein the method comprises the following steps: the method for constructing the fucoidin-liver injury protein interaction network and predicting the core target point comprises the steps of adopting a STRING database http:// STRING-db.org/Multiple Proteins term lower limit species to be human Homo sapiens, uploading the intersection target point in the step 2) to construct the protein interaction network, analyzing and setting a minimum interaction value of 0.09 to obtain a target interaction network and related data, carrying out a tsv format, introducing the tsv data into Cytoscape software, analyzing a Degree of freedom median and a maximum Degree of freedom topological parameter in the network by using a NetworkAnalyzer module, screening the core target point according to the Degree value, wherein the upper limit of a screening range is the maximum Degree value in the topological data, and the lower limit is two times the Degree of freedom median, and filtering to obtain the fucoidin-liver injury core target point.

5. The method for analyzing the mechanism of action of fucoidan against liver injury based on cyber pharmacology according to claim 1, wherein the method comprises the following steps: uploading the fucoidin-liver injury core target in the step 3) to a DAVID database http:// david.ncifcrf.gov/home.jsp, analyzing the interaction relationship between the fucoidin-liver injury core target and GO biological functions and a KEGG channel, realizing visualization operation by means of an R language 3.6.1 'GOPLOT' R packet to obtain a corresponding histogram and a bubble map, introducing the data into a credit-producing online tool to obtain a Circos circle map, and obtaining the biological function distribution and participation channel information of the key core target according to the graph.

[ technical field ] A method for producing a semiconductor device

The invention belongs to the technical field of system bioinformatics, and particularly relates to a method for analyzing an anti-liver injury action mechanism of fucoidin based on network pharmacology.

[ background of the invention ]

The modern high-tech technology accelerates the development of people on the marine environment, and the abundant species resources of the sea gradually become unique sources of daily supplies such as foods, cosmetics, health care products and the like, wherein the natural active ingredients of the sea are more of great concern in the field of drug research and development. Fucoidan (Fucoidan) is a complex sulfate polysaccharide, and is mainly derived from brown algae, and can also be extracted from invertebrate (such as sea cucumber and sea urchin) or sea grass. Fucoidan mainly comprises L-fucose and sulfate group, and contains monosaccharides such as rhamnose, arabinose, glucose, and galactose. The molecular weight of fucoidan is from 1000Da to 10000Da, and the specific composition and structure of fucoidan usually vary according to algae species, growth environment, harvest season, extraction and separation method, so the biological activity of fucoidan with different composition and structure also varies. Since Kylin is discovered for the first time in 1931, with the continuous and intensive research, various pharmacological activities of fucoidan are gradually known. At present, the most prominent pharmacological action of fucoidan is antitumor activity. Researches find that the fucoidin can reduce the migration and diffusion capacity of tumor by blocking the adhesion and invasion of tumor cells to endothelial cells, regulate the expression of key proteins of cell growth and apoptosis and inhibit the abnormal proliferation of cells, thereby playing the role of resisting tumor. The fucoidin can adjust the intestinal flora balance of colon cancer rats induced by 1, 2-dimethylhydrazine through a 16S rRNA high-throughput sequencing technology, so that an experimental basis is provided for preventing the colon cancer rats from happening and developing. In addition, the antioxidation of fucoidan is also a research hotspot. Fucoidin obtained by different extraction methods has different antioxidant activities, and the fucoidin extracted from Ascophyllum nodosum by adopting a microwave extraction method has higher DPPH (free radical) removing capability than the fucoidin extracted at the extraction temperature of 120 ℃ or 150 ℃ under the condition of 90 ℃. For pathological injury induced by obesity, fucoidan not only can relieve accelerated and aggravated obesity ischemic brain injury by relieving severe oxidative injury caused by obesity, but also can reduce the level of blood vessel growth factor-B, and further reduce hypertension of obese mice. It is worth noting that the unique pharmacological action of fucoidan is closely related to its source. New method for purifying Antarctic HairpusFucoidin (F-4) has the effect of promoting the growth of leukocytes in vivo, and the action mechanism of the fucoidin can be related to activating hematopoietic cells and regulating a bone marrow hematopoietic microenvironment. Fucoidan extracted from Laminaria maxima of south Africa is a potent inhibitor of alpha-glucosidase, IC thereof₅₀The range of the drug is obviously lower than that of acarbose, and the drug is one of potential drugs for treating type 2 diabetes. Besides being used as a medicine for treating diseases such as tumor, diabetes and the like, fucoidan is also an ideal material for encapsulating active ingredients and is used for improving the treatment effect of the medicine. Compared with the common quininacrine preparation, the quininacrine fucoidan nano-particle prepared by taking fucoidan as an active targeting body has the advantages of enhanced anticancer activity, weakened hepatotoxicity and higher animal survival rate. Considering the endothelial cell adhesion and anticoagulation effects of fucoidin, the fucoidin and polyvinyl alcohol (PVA) are co-crosslinked by Yao and colleagues to prepare the blood vessel transplantation material, and the in vitro experiment results show that the fucoidin modified polyvinyl alcohol can effectively reduce the platelet adhesion and reduce the thrombosis. Fucoidin promotes endothelialization of PVA through modifying the surface of the PVA, and strengthens the adhesiveness of PVA cells, thereby achieving the purpose of blood vessel transplantation. At present, fucoidin has good drug development prospect due to the advantages of various pharmacological activities, no toxicity, degradability and biocompatibility.

The liver, as the largest parenchymal organ of the human body, mainly plays a role in substance metabolism and is an important role in maintaining normal life activities of the organism. The incidence of liver diseases is getting worse and worse all over the world, and the liver diseases are still a big problem for doctors all over the world. Viral infection, metabolic diseases, drinking and the like can cause serious liver diseases such as fatty liver, liver cirrhosis, hepatitis, liver cancer and the like, and finally cause liver injury. Recent studies have shown that fucoidan has great potential in the treatment of liver damage. The oral administration of fucoidin 50mg/kg per day in rats with liver injury can reduce inflammatory factor level, inflammatory cell infiltration and collagen fiber deposition, and inhibit hepatic fibrosis. Fucoidan also reduces aflatoxin B1-induced liver mitochondrial dysfunction, oxidative damage, and is believed to have prophylactic and therapeutic effects on food contamination poisoning. Similarly, fucoidin is added into the liver preservation solution of the isolated rat, so that the oxidative stress and apoptosis pathway of cells can be adjusted, and the functional integrity of the liver is maintained, thereby achieving the purpose of reducing the ischemia reperfusion injury of the transplanted liver. Therefore, the anti-liver injury effect of fucoidin can be the co-coordination of multiple mechanisms such as anti-inflammation, anti-oxidation, anti-apoptosis and the like.

Despite the various pharmacological activities of fucoidan, most of the studies are still preclinical studies and clinical trials are deficient. In order to reveal the mechanism of fucoidan for resisting liver injury, research and determination of fucoidan-liver injury action target points are necessary, a basic theoretical basis is provided for the clinical drug development of fucoidan, and a new idea is provided for the research of liver injury related drugs.

[ summary of the invention ]

Aiming at the defects that most of researches on fucoidin with various pharmacological activity effects in the prior art are preclinical researches and lack of clinical trials, the invention provides a method for analyzing the mechanism of the fucoidin for resisting liver injury based on network pharmacology, which is a method for disclosing the molecular mechanism of the fucoidin for resisting liver injury based on network pharmacology, and the method comprises the steps of mining the common target of the two on the basis of network pharmacology, constructing a target-protein interaction network, analyzing the relevant passage of fucoidin for treating liver injury by combining bioinformatics and providing reference for subsequent researches.

The purpose of the invention is realized by the following technical scheme:

a method for analyzing the mechanism of action of fucoidin against liver injury based on network pharmacology comprises the following steps:

1) fucoidin target acquisition and correction: using a drug online database to obtain fucoidan action targets, and correcting all the targets in a protein database to match with related gene symbols;

2) obtaining a liver injury target spot and identifying an intersection target spot: searching and searching for a Liver injury candidate target point in Genecard and DisGeNET databases by taking 'Liver injury' as a keyword, and then performing intersection mapping with the fucoidin action target point obtained in the step 1) to obtain a fucoidin anti-Liver injury action target point;

3) constructing a fucoidin-liver injury protein interaction network and predicting a core target: analyzing the intersection target in the step 2) by adopting a STRING database to obtain a target protein interaction relational graph (PPI network) and tsv data thereof, analyzing the tsv data (network nodes) by using a networkAnalyzer module in Cytoscape3.7.1 software to obtain the median of freedom and the maximum degree of freedom, wherein the upper limit and the lower limit of the screening of the core target are twice the maximum degree of freedom and the median of freedom respectively, and the target of which the topological parameters are contained in the upper limit and the lower limit ranges is taken as the fucoidan-liver injury core target;

4) core target biological function and enrichment analysis discuss the mechanism of action of fucoidin against liver injury: a gene ontology database (GO) is used for analyzing the biological functions of fucoidin in resisting liver injury, a Kyoto gene and genome encyclopedia database (KEGG) is used for revealing a fucoidin signal channel in resisting liver injury, and finally, a corresponding bubble diagram, a histogram and a Circos circle diagram are output;

5) the construction and visualization of a 'drug-target-gene ontology function-pathway-disease' network: constructed according to the results of step 4) using cytoscape 3.7.1.

In the invention:

the drug online database in the step 1) is selected from Swiss Target Prediction, PharmMapper, Batman and drug bank; the protein database is selected from Uniprot; the correction to match the relevant gene symbols is carried out by selecting a verified (reviwed) Human source target (Human) as a screening condition, and converting the name of the sorted target into a gene name to obtain a corrected medicine target.

The step 2) of carrying out intersection mapping with the fucoidan action target spot obtained in the step 1) is to use a letter on-line tool for correcting the fucoidan action target spot in the step 1) of finishing the liver injury target spot

(http:// bioinformatics. psb. element. be/webtools/Venn /) is processed with the Venn map (Venn) map intersection.

The method for constructing the fucoidin-liver injury protein interaction network and predicting the core target point comprises the steps of adopting an STRING database (http:// STRING-db. org) Multiple Proteins to define a lower limit species as a human (Homo sapiens), uploading the intersection target point in the step 2) to construct the protein interaction network, analyzing and setting a minimum interaction value of 0.09 to obtain a target interaction network and related data (tsv format), introducing the tsv data into Cytoscape software, analyzing topological parameters such as a median Degree of freedom and a maximum Degree of freedom in the network by using a networkAnalyzer module, screening the core target point according to the Degree value, wherein the upper limit of a screening range is the maximum Degree value in the topological data, the lower limit is twice the median Degree of freedom, and filtering to obtain the fucoidin-liver injury core target point.

And 3) the fucoidin-liver injury protein interaction network is used for establishing a fucoidin treatment target protein and a liver injury disease target protein by using a database platform and displaying an interaction relation between the two protein targets.

The core target biological function and enrichment analysis of the step 4) discuss the mechanism of fucoidin action against liver injury, and the method is characterized in that the fucoidin-liver injury core target in the step 3) is uploaded to a DAVID database (http:// DAVID. ncifcrf. gov/home. jsp), the interaction relation between the fucoidin-liver injury core target and GO biological function and a KEGG channel is analyzed, the visualization operation is realized by means of R packages such as R language (3.6.1) "GOPLOT" and the like, a corresponding bar graph and a bubble graph are obtained, meanwhile, data are led into a letter on-line tool to obtain a Circos circle graph, and the key core target biological function distribution and participation channel information can be obtained according to the graph.

The construction and visualization of the 'drug-target point-gene ontology function-pathway-disease' network in the step 5) is to simplify the relation between the target point and the biological function and the signal pathway involved in the fucoidan anti-liver injury, and the construction of the 'drug-target point-gene ontology function-pathway-disease' network is carried out by using Cytoscape software, and the network can concisely reveal the molecular action mechanism of the fucoidan anti-liver injury.

Compared with the prior art, the invention has the following advantages:

1. the invention relates to a method for analyzing an anti-liver injury action mechanism of fucoidin based on network pharmacology, which belongs to the field of system bioinformatics, constructs a drug-disease network through a cloud platform and big data analysis, and effectively reveals drug treatment disease targets and action principles.

2. The invention relates to a method for analyzing an anti-liver injury action mechanism of fucoidin based on network pharmacology, which is used for researching the anti-liver injury mechanism of the fucoidin for the first time by adopting the network pharmacology method; the result of the invention can be used as a breakthrough point for follow-up research and verification, which provides a value reference for comprehensively understanding the pathogenesis of related liver diseases and the curative effect target of fucoidin.

3. The invention discloses a method for analyzing an anti-liver injury action mechanism of fucoidin based on network pharmacology, which firstly reveals a key target of the anti-liver injury of the fucoidin and a related biological process. The invention adopts a network pharmacology method to determine the fucoidin-liver injury action target point, provides a basic theoretical basis for the clinical medication development of the fucoidin and provides a new idea for the research of liver injury related drugs.

[ description of the drawings ]

FIG. 1 is a diagram of fucoidan anti-liver injury intersection target Venn-protein PPI in an example of the present invention;

FIG. 2 is a fucoidan-liver injury core target plot in an example of the present invention;

FIG. 3 is a bar graph of the GO biological function of fucoidan against liver damage in an embodiment of the invention;

FIG. 4 is a graph of fucoidan anti-liver injury GO biofunctional bubbles in an example of the invention;

FIG. 5 is a graph of the GO cycle with fucoidan biofunctionality against liver damage in an embodiment of the invention;

FIG. 6 is a bar graph of fucoidan anti-liver injury KEGG pathway enrichment in an embodiment of the invention;

FIG. 7 is a graph of fucoidan anti-liver injury KEGG pathway enriched bubbles in an example of the present invention;

FIG. 8 is a graph of the fucoidan anti-liver injury KEGG pathway enrichment circles in an example of the invention;

FIG. 9 is a graph of the fucoidan-target-GO-KEGG-liver injury network in the example of the present invention;

FIG. 10 is a flow chart of the procedure of the anti-liver injury study of fucoidan according to the example of the present invention.

[ detailed description ] embodiments

The following examples are provided to further illustrate the embodiments of the present invention.

Example (b):

a method for analyzing the mechanism of action of fucoidin against liver injury based on network pharmacology comprises the following steps:

1. method step

1.1 fucoidan target acquisition and correction

Taking the English name of Fucoidan as a search term, respectively entering drug databases (Swiss Target Prediction, PharmMapper, Batman and drug bank) to comprehensively search the Fucoidan drug action Target, collecting the Target obtained by searching each database, removing repeated items, and performing Target correction by adopting a Uniprot protein database: selecting a verified (reviewed) Human source target (Human) as a screening condition, and converting the name of the sorted target into a gene name to obtain a corrected drug target;

1.2 liver injury target acquisition and intersection target identification

Searching for the disease target by using a Liver injury English name 'Liver injury' as a keyword, searching for the Liver injury target by using a disease database (GeneCard and DisGeNET), and then performing target duplication removal operation; performing Venn diagram (Venn) mapping intersection treatment on the prepared liver injury target and the 1.1 corrected fucoidin target by using a letter on-line tool (http:// bioinformatics. psb. element. be/webtools/Venn /), so as to obtain a fucoidin anti-liver injury intersection target gene;

1.3 construction of fucoidin-liver injury protein interaction network and prediction of core target

The method comprises the following steps of defining the lower limit of an STRING database (http:// STRING-db. org /) Multiple Proteins as a human (Homo sapiens), uploading a 1.2 intersection target to construct a protein interaction network, analyzing and setting the lowest interaction value of 0.09 to obtain a target interaction network and related data (tsv format), then importing the tsv data into Cytoscape software, analyzing topological parameters such as the median of freedom Degree and the maximum freedom Degree in the network by using a NetworkAnalyzer module, screening a core target according to the Degree value, wherein the upper limit of a screening range is the maximum Degree value in the topological data, the lower limit is twice the median of the freedom Degree, and filtering to obtain a fucoidan-liver injury core target;

1.4 core target biological function and enrichment analysis discussion of mechanism of action of batholith polysaccharide against liver injury

Uploading 1.3 fucoidin-liver injury core target to a DAVID database (http:// DAVID. ncifcrf. gov/home. jsp), analyzing the interaction relationship between the fucoidin-liver injury core target and GO biological function and a KEGG channel, realizing visual operation by means of R packages such as R language (3.6.1) "GOPLOT" and the like to obtain a corresponding histogram and a bubble diagram, simultaneously introducing data into a letter on-line tool to obtain a Circos circle diagram, and obtaining the biological function distribution and participation channel information of the key core target according to the diagram;

1.5 construction and visualization of drug-target-gene ontology function-pathway-disease network

In order to simplify the relation between the fucoidin and the biological function and the signal path related to the anti-liver injury of the fucoidin, Cytoscape software is utilized to construct a 'medicine-target spot-gene body function-path-disease' network, and the network can concisely disclose the molecular action mechanism of the fucoidin for resisting the liver injury.

2. Results

2.1 drug disease target screening and aggregation of the two intersecting targets

In the research, drug targets are searched through databases such as Swiss Target Prediction, Drugbank and the like, 200 fucoidan action targets are obtained in total through correction of a Uniprot database after repeated items are removed, 7268 liver injury genes are collected through a Genecard database and a DisGeNET database, and 180 fucoidan-liver injury intersection targets are obtained by mapping and intersecting the two targets: MMP2, CD44, CXCR4, GP1BA, NOS2, GDA, SELL, TNFRSF9, BCHE, BCL2, FGF1, CIITA, NEU1, PTPN2, PPARGC1A, TNFAIP6, CYP19A1, LDLR, CA9, XIAP, OLR1, AGER, CD83, VEGFA, MME, GSK3B, SMURF2, FPR1, PPARD and the like, and is shown in detail in FIG. 1.

2.2 fucoidan-hepatic injury PPI network and core target aggregation

The method comprises the steps of importing 180 common targets into a STRING database, setting the score to be greater than 0.09 to obtain a fucoidin anti-liver injury PPI network (figure 1), expressing the relationship between two nodes by edges, connecting a plurality of targets by a plurality of edges to show the complex relationship between the targets, calculating fucoidin-liver injury protein interaction network node parameters through Cytoscape software, knowing the median of the degree of freedom of the fucoidin acting on the liver injury target network 37.345, and the maximum degree of freedom of 110, wherein the screening range of the core targets is 75-110, and finally obtaining 7 core targets, namely ALB, CXCL8, TP53, TNF, SRC, IL6 and VEGFA, wherein relevant detailed information is shown in a table 1 and a figure 2.

TABLE 1 fucoidan-liver injury core target information

Tab.1 The hub biotargets of information for Fucoidan against Liver injury

2.3 fucoidin-liver injury key target GO functional analysis and KEGG enrichment analysis

To fully explore and discuss the potential pharmacological mechanism of fucoidan against liver injury, GO and KEGG analysis were performed on the drug-disease intersection target in this study.

2.3.1GO bioanalysis

GO biological information analysis was performed on fucoidan-liver injury key targets using DAVID database to obtain corresponding graphs (table 2, fig. 3, fig. 4 and fig. 5), from which it can be seen that fucoidan anti-liver injury involves biological processes: a positive regulation of peptidyl-tyrosine phosphorylation, cellular response to lipopolysaccharide, negative regulation of apoptotic process, positive regulation of ERK1 and ERK2 cassette (ERK1 and ERK2 cascade), and the like; the cell components involved are: extracellular space, extracellular region, platelet alpha granule lumen, external side of plasma membrane; the related molecular functions are: identification protein binding (same protein binding), cytokine activity (cytokine activity), protein binding (protein binding), transcription regulation region DNA binding (transcription regulatory region DNA binding), and the like.

TABLE 2 Key target GO bioanalysis

Tab.2 GO biological analysis of hub targets

2.3.2 KEGG pathway enrichment analysis

The KEGG pathway enrichment analysis results showed that a total of 16 signaling pathways were associated with fucoidan anti-liver injury core targets (table 3, fig. 6, fig. 7 and fig. 8), including: VEGF signaling pathway (vascular endothelial growth factor signaling pathway), toll-like receptor signaling pathway (toll-like receptor signaling pathway), TNF signaling pathway (tumor necrosis factor signaling pathway), thyroid hormone signaling pathway (thyroid hormone signaling pathway), sphingolipid signaling pathway (sphingolipid signaling pathway), NOD-like receptor signaling pathway (NOD-like receptor signaling pathway), NF-kappa B signaling pathway (nuclear factor kappa B signaling pathway), cytokine-cytokine receptor interaction, and the like.

TABLE 3 Key target KEGG pathway enrichment analysis

Tab.3 KEGG pathway of hub targets enrichment analyses

2.4 construction and visualization of "drug-target-Gene ontology function-pathway-disease" network

After experimental data are analyzed, network visualization of 'fucoidin-target-GO-KEGG-liver injury' is carried out, and the result is shown in figure 9, the figure mainly comprises a fucoidin anti-liver injury core target, related GO functions (biological processes, cell components and molecular functions) and KEGG channels, and as can be seen from the figure, the fucoidin regulates a plurality of signal channels to play a role in resisting liver injury by acting on a plurality of core targets and a plurality of biological processes.

3. Discussion of the related Art

The invention constructs a drug-disease network and performs enrichment analysis on key target passage by combining a network pharmacology method and a plurality of database platforms of drugs, diseases, proteins and the like, systematically discusses the mechanism of the fucoidin for resisting liver injury, and researches show that the fucoidin has the characteristics of multi-target and multi-passage action for resisting liver injury.

The results show that seven core targets are shared, and ALB, CXCL8, TP53, TNF, SRC, IL6 and VEGFA are involved in the action process of fucoidan for resisting liver injury, and the targets are possible to be effective pharmacological targets of the fucoidan for resisting liver injury. Hepatitis is a major cause of liver injury, and the long-term inflammatory response of hepatocytes can progress to hepatic fibrosis and cirrhosis. Cells in the initial stage of inflammation release a series of inflammatory cytokines and chemokines through a nuclear factor kB signal channel to activate immune cells, such as neutrophils, phagocytes, T cells and the like, so that the immune cells are induced to be aggregated at an inflammation part to further release the inflammatory factors, and a large amount of inflammatory factors and inflammatory cell infiltration can continuously trigger inflammatory stress to cause in-vivo inflammatory regulation disorder. TNF and IL are key members of the inflammatory factor family and are divided into different subtypes such as TNF-alpha, TNF-beta, IL-1, IL-6, IL-8 and the like. Research reports that the anti-inflammatory effect of fucoidin is related to the reduction of the level of inflammatory factors, the levels of TNF-alpha and IL-6 in mice with C57BL/6 liver injury treated by fucoidin are obviously reduced, and the condition of liver inflammation is effectively relieved. In addition, fucoidan can act on Toll-like receptor of bronchial epithelial cell, and inhibit expression of TNF-alpha, IL-6 and CXCL8, thereby relieving airway inflammation^[19]. ALB synthesized by liver cells has the capability of binding ligand, and can store and transport internal and external compounds. The protein has anti-inflammatory effect, and helps to stabilize endothelial cells and maintain capillary permeability. It is common to determine clinically the ALB level indicative of abnormal liver changes. In addition to inflammatory reactions, liver cancer characterized by uncontrolled proliferation of hepatocytes, apoptosis and necrosis is also a risk of liver injuryFactors. TP53 is a key cancer gene, and its regulation can affect tumor formation and development by regulating its level to regulate the expression of upstream and downstream signal channels. Fucoidin can reduce the activity of HCT116 colon cancer cells and inhibit the tumorigenicity of the cancer cells to nude mice by promoting the expression of P53, thereby showing strong anticancer effect. SRC is essentially a tyrosine kinase, activated SRC plays an important role in the development of multiple tumorigenesis and causes liver damage, and it also mediates inflammation in vivo through covalent modification of the transactivation of EGFR-mediated ERK1/2 pathway. Vascular growth factor a (vegfa) plays a central role in angiogenesis, promoting endothelial cell proliferation. For liver injury caused by fatty liver, VEGFA combined with soluble vascular endothelial growth factor receptor 1 treatment can protect fatty liver through VEGFR2-PI3K/Akt pathway. As a main medium of angiogenesis, VEGFA can also stimulate solid tumor vasodilatation, enhance the microenvironment function of tumor blood vessels, and promote tumor formation. In order to further analyze the mechanism of the fucoidin anti-liver injury molecule, the further combination of GO and KEGG enrichment analysis results show that the fucoidin mainly acts on the seven targets, regulates and controls biological functions such as peptidyl tyrosine phosphorylation, ERK1 and ERK2 cascade reaction, protein combination and the like, influences the signal path expression of a vascular endothelial growth factor signal path, a tumor necrosis factor signal path, a nuclear factor kB signal path, cytokine-cytokine receptor interaction and the like, and plays a role in anti-liver injury.

4. Conclusion

In conclusion, the invention determines the specific biological pharmacological target, biological function and related signal pathways of the fucoidin for resisting liver injury through network pharmacology. The network pharmacology finds that relevant targets of fucoidin for resisting liver injury are as follows: ALB, CXCL8, TP53, TNF, SRC, IL6 and VEGFA, which provide a new methodology for further researching related pharmacological molecular mechanisms of fucoidin, and fucoidin has potential therapeutic effect on liver injury.

The above description is intended to describe in detail the preferred embodiments of the present invention, but the embodiments are not intended to limit the scope of the claims of the present invention, and all equivalent changes and modifications made within the technical spirit of the present invention should fall within the scope of the claims of the present invention.