Method for exploring ascending of platycodon grandiflorum guiding medicine based on network pharmacology

Abstract

The invention belongs to the field of traditional Chinese medicines, and particularly relates to a method for exploring ascending of a radix platycodonis guide medicine based on network pharmacology, which explores a mechanism of a radix platycodonis'guide medicine ascending 'effect from a molecular level by using a network pharmacology method. The method comprises the following steps: searching and collecting compounds contained in platycodon grandiflorum and corresponding targets through a database, constructing and analyzing a target network, determining important targets and sub-modules in the network, carrying out function enrichment analysis on the important targets and the sub-modules, and researching an action mechanism of'guiding medicine upward 'of platycodon grandiflorum. Correct and objective understanding of the effect of guiding the medicine to move upwards is beneficial to better guidance of clinical practice.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine, and particularly relates to a method for exploring the upward movement of Platycodon grandiflorum-induced drugs based on network pharmacology. Background Art

[0002] The theory of guiding the meridians is a medication experience developed through long-term clinical practice, building upon the theory of meridians in Traditional Chinese Medicine. However, it extends beyond the specific meridians of a single drug. Guiding the meridians, also known as "guiding the meridians and sending messengers," is a key component of prescription composition and falls within the scope of medication. "Yin" means to introduce or guide, while "jing" refers to specific organs, meridians, or diseased areas. Guiding drugs refer to certain herbs that can direct the medicinal properties of other herbs to the affected area or meridians, acting as a "guide."

[0003] Platycodon grandiflorum (Jacq.) A.DC., a member of the Campanulaceae family, has been used in China for over a thousand years. It possesses anti-inflammatory and expectorant properties and is clinically used to promote lung function, relieve sore throat, treat lung abscesses, relieve cough and expectoration, promote blood circulation, dissipate lung qi and dispel dampness, and transport herbs upward. The earliest recorded text appears in the Shennong Bencao Jing (Shen Nong's Classic of Materia Medica), which states, "It is pungent and slightly warm. It is primarily used to treat chest and flank pain, abdominal distension, rumbling bowels, and panic and palpitations." During the Song, Jin, and Yuan dynasties, the book Zhenzhu Nang Yaoxing Fu (The Properties of Herbs) states, "It is bitter, pungent, slightly warm, and slightly toxic. It is an ascending agent, the yang within the yin. It has four uses: to relieve sore throat and nasal congestion; to promote diaphragmatic qi and treat lung abscesses; to act as a vessel for other herbs; and to guide the lung meridians." Through clinical application by physicians throughout history, Platycodon grandiflorum's clinical use has gradually expanded, and its efficacy has been refined. Its use as a channeling agent, in particular, has been widely recognized by physicians both ancient and modern. While numerous compound preparations containing Platycodon grandiflorum are available clinically, no specific target for its "channeling agent" has been identified. By studying the mechanism of action of Platycodon grandiflorum in channeling agent, a correct and objective understanding of its efficacy will help better guide clinical practice.

[0004] The role of Platycodon grandiflorum's "guiding medicine upward" effect in compound preparations

[0005] The "Shenling Baizhu San" in the "Taiping Huimin Hejijufang" is used to treat spleen and stomach deficiency, poor appetite, fatigue and lack of energy, fullness and suffocation, heart palpitations and shortness of breath, vomiting and diarrhea, and damage to the middle and lack of heat. Long-term use nourishes qi and spirit, invigorates the spleen and brightens the complexion, and wards off evil spirits. Most physicians believe that Platycodon grandiflorum guides other herbs upward to the lungs. As stated in the "Yi Yi Bing Shu," "Medicines that guide the meridians are like a guide for a person who does not know the way." As a guide, Platycodon grandiflorum is known as a "boat and navigator" because it can guide herbs upward to higher places, guiding and propelling other herbs to treat upper body ailments. This use of Platycodon grandiflorum as a boat and navigator originated with Zhang Yuansu's discussion: "Plantococcus grandiflorum clears lung qi and soothes the throat. Its white color makes it a lung guide, and together with licorice, it serves as a boat and navigator."

[0006] "Tianwang Buxin Dan" is composed of 14 Chinese herbs, namely ginseng, salvia miltiorrhiza, Scrophularia ningpoensis, white pueraria, schisandra chinensis, polygala tenuifolia, platycodon grandiflorum, angelica root, asparagus cochinchinensis, ophiopogon japonicus, cypress seed, jujube seed, raw rehmannia root, and cinnabar. Its effects are to nourish the heart, calm the mind, nourish yin, and clear away heat. It is commonly used in the clinic to treat various diseases caused by deficiency of yin and blood, such as palpitations, insomnia, fatigue, nocturnal emission, forgetfulness, dry stool, mouth sores, red tongue with little coating, and weak pulse. This prescription uses raw rehmannia root and Scrophularia ningpoensis as the main ingredients. Raw rehmannia root nourishes yin and cools blood, replenishes kidney water to clear heart fire, and Scrophularia ningpoensis nourishes yin and produces body fluid, so that the mind is not disturbed by the deficiency fire. The two herbs are used together as the main ingredients to replenish water to control fire, so that kidney water rises and heart fire is not too strong, and heart fire falls and kidney water is not cold, which is the idea of water and fire being in harmony[4]. This prescription incorporates the auxiliary herbs of Chinese angelica, salvia miltiorrhiza, ginseng, huo zhu, cypress seed, and polygala tenuifolia. Angelica tonifies and invigorates blood circulation, while salvia miltiorrhiza clears stagnant heat and relieves restlessness. Together, they nourish the blood, nourish the heart, and relieve restlessness. Ginseng promotes fluid production and invigorates qi, calming the mind. huo zhu strengthens the spleen and soothes the mind, while polygala tenuifolia calms the mind. Cypress seed nourishes the heart and spleen, collectively tonifying qi and invigorating the spleen, calming the mind and tranquilizing the mind. Schisandra chinensis, spinach seed, asparagus cochinchinensis, and ophiopogon japonicus serve as adjuvants. Asparagus cochinchinensis and ophiopogon japonicus clear upward-moving internal heat, while schisandra chinensis and spinach seed restrain depleted heart qi, calming the mind. Platycodon grandiflorum is used as a guiding herb to ensure the medicinal effects reach the heart and ease their downward movement to the kidneys. In his analysis of this prescription in his book "Famous Physician Prescriptions," Ke Qin of the Qing Dynasty wrote, "Plantago grandiflora acts as a vessel and polygala tenuifolia as a guide, allowing the other herbs to enter the heart and soothe the mind."

[0007] Xuefu Zhuyu Decoction is primarily used to treat chest blood retention syndrome. This formula is a combination of Taohong Siwu Decoction and Sini San, with Platycodon grandiflorum and Achyranthes bidentata. Taohong Siwu Decoction promotes blood circulation, dissipates retention, and nourishes the blood. Sini San soothes the liver and regulates qi, while the addition of Platycodon grandiflorum guides the herbs upward to the chest (the blood palace). Achyranthes bidentata guides blood stasis downward, unblocking the blood vessels. This is a commonly used formula for promoting blood circulation, dispelling retention, and promoting qi and relieving pain. Regarding the role of Platycodon grandiflorum in this formula, the textbook "Pharmacology of Prescriptions" explains that "Plantagecodon grandiflorum invigorates the lungs and carries the herbs upward. Combined with Ji Ke, it raises and lowers qi in the upper burner and relieves chest tightness. Achyranthes bidentata, in particular, unblocks the blood vessels and draws blood downward. These interactions activate blood circulation and qi, dissipating retention heat and relieving liver depression, leading to spontaneous resolution of all symptoms." This clearly indicates that Platycodon grandiflorum's "guiding meridian" function is to "open and promote lung qi and carry the herbs upward." Ding proposed that according to the Ben Jing (Classic Classic of Materia Medica), Platycodon grandiflorum can treat "chest and flank pain resembling a knife stab." This pain, apparently due to blood retention, is clearly caused by blood retention, and Platycodon grandiflorum is the primary treatment, demonstrating its true blood-regulating properties. There are two main reasons for its use. First, Platycodon grandiflorum is good at regulating Qi, and Qi is the commander of blood. When Qi moves, blood moves. Stagnation of Qi causes blood to stagnate. To treat blood stagnation and blood stasis, it is necessary to regulate Qi, and promoting Qi helps blood circulation. Second, Platycodon grandiflorum itself has the power to activate blood circulation. This is why Platycodon grandiflorum is effective in treating "pain like a knife" in Ben Cao. Therefore, Platycodon grandiflorum is often used in combination with prescriptions for the treatment of angina pectoris caused by coronary heart disease in clinical practice, and it always has a good effect.

[0008] Modern pharmacology's understanding and research on "drug delivery" in traditional Chinese medicine

[0009] In 1997, Hu Yizhong also conducted preliminary research on Platycodon grandiflorum's ability to "direct drugs upward." A study of the erysipelas formula "Xiaoyan Tang" (Xiaoyan Tang) using Platycodon grandiflorum and Achyranthes bidentata in the treatment of upper and lower limb erysipelas, respectively, demonstrated that Platycodon grandiflorum directed drugs upward, resulting in significantly higher blood concentrations in ascending blood flow than in descending blood flow. Other researchers have experimentally revealed Platycodon grandiflorum's "directing" function in Bai San (Bai San). Removing Platycodon grandiflorum only cleared ascites, but not pleural effusions. Through literature research, Duan Zongyi believed that the solubilization effect determined by the surface activity of Platycodon grandiflorum soap test and the influence on cell membrane permeability are the intrinsic basis of Platycodon grandiflorum's meridian-guiding effect; Yu Wenhai concluded through literature research that the "hydrophiles" and "lipophiles" contained in meridian-guiding drugs may be different from non-meridian-guiding drugs in molecular structure and particle size, and therefore have the function of guiding the effective ingredients of other drugs to the required parts of the body; Li Wai et al. used the levels of cyclic nucleotides (cAMP, cGMP) in rat brain, heart, and lung tissues as indicators to study the role of Platycodon grandiflorum in "carrying drugs upward" in Tianwang Buxin Pills, and concluded that the role of Platycodon grandiflorum in "carrying drugs upward" in Tianwang Buxin Pills may be closely related to the lungs. These studies have actively explored the "meridian-guiding effect" of Platycodon grandiflorum.

[0010] Li Yinglun conducted a modern experimental study on the "drug-carrying" effect of Platycodon grandiflorum using pharmacokinetic and tissue dynamics methods. He investigated the effect of Platycodon grandiflorum on roxithromycin concentration in the lungs after combining it with roxithromycin. The results showed that Platycodon grandiflorum's "channel-guiding" effect increased roxithromycin concentration in lung tissue. It also significantly affected the pharmacokinetic properties of levofloxacin in healthy chickens and the distribution of drugs. It also significantly affected the pharmacokinetic properties of florfenicol in healthy rabbits. The material basis for Platycodon grandiflorum's "drug-carrying" effect is Platycodon grandiflorum total saponins. He first proposed that Platycodon grandiflorum total saponins inhibit the activity and expression of P-gP in the small intestine and lung tissues as the mechanism of Platycodon grandiflorum's "channel-guiding" effect, confirming the existence of this "drug-carrying" effect.

[0011] The theory of guiding meridians is the essence of Traditional Chinese Medicine. In-depth research on this theory not only confirms the rationality and practicality of TCM theory and corrects misconceptions about it, but also helps us scientifically organize traditional theories, making them easier to pass down and more widely understood and accepted. While Platycodon grandiflorum's "guiding herbs upward" effect has been demonstrated through compound preparations and modern pharmacological experiments, its mechanism remains unclear.

[0012] Traditional Chinese medicine (TCM) possesses multiple components, multiple targets, and diverse regulatory mechanisms, embodying a vast amount of information. Since its initial introduction by HOP-KINS in 2007, network pharmacology has been widely applied to elucidate the pharmacological mechanisms and investigate the toxicity mechanisms of TCM, transforming the traditional "one drug, one target, one disease" approach to new drug development. Based on the "disease-gene-target-drug" interplay, network pharmacology analyzes the relationship between drug molecules and disease networks from the perspective of overall biological network stability, using network targets as a starting point.

[0013] Based on this, the present invention is proposed. Summary of the Invention

[0014] The purpose of the present invention is to provide a method for exploring the upward movement of Platycodon grandiflorum based on network pharmacology to solve the above problems.

[0015] A method for exploring the upward movement of Platycodon grandiflorum based on network pharmacology comprises the following steps:

[0016] Step 1: Collection and screening of chemical components from Platycodon grandiflorum

[0017] Platycodon grandiflorum is screened to find effective compounds that meet the conditions and obtain chemical components that meet the conditions;

[0018] Step 2: Obtaining target genes corresponding to effective compounds of Platycodon grandiflorum

[0019] Through the effective compounds of Platycodon grandiflorum, the corresponding gene names are obtained, and one target corresponds to one gene name;

[0020] Step 3: Obtaining disease genes

[0021] By using a disease associated with Platycodon grandiflorum as a keyword search to obtain corresponding known targets, the corresponding targets are analyzed;

[0022] Step 4: Constructing common disease targets

[0023] Construct the intersection of Platycodon grandiflorum and disease targets to obtain the common targets of Platycodon grandiflorum and diseases;

[0024] Step 5: Construct a network diagram of drugs, active ingredients, and disease targets

[0025] By searching for effective gene targets, a network diagram of drugs, active ingredients and disease targets is constructed;

[0026] Step 6: Construct a network diagram of protein-protein interactions between Platycodon grandiflorum and common targets

[0027] By importing the target proteins of the common targets obtained through screening into the database, a network diagram of protein-protein interactions between Platycodon grandiflorum and the common targets was constructed;

[0028] Step 7: GO functional enrichment analysis and KEGG pathway enrichment analysis

[0029] The disease target intersection data was uploaded to the David platform, and the data was visualized to obtain a bubble chart. Then, based on the bubble chart and combined with the existing information, the mechanism of Platycodon grandiflorum leading the drug upward was analyzed.

[0030] As a further improvement, Platycodon grandiflorum was screened, and 130 effective compounds were screened out, of which 22 met the requirements.

[0031] Further improvements were made, and 7 target genes corresponding to 7 effective compounds of Platycodon grandiflorum were obtained based on 22 effective ingredients.

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] This study uses network pharmacology to explore the molecular mechanism of Platycodon grandiflorum's "drug-inducing" effect. By searching a database to identify compounds and their corresponding targets, a target network was constructed and analyzed, identifying key targets and submodules within the network. Functional enrichment analysis was then performed on these key targets and submodules to investigate the mechanism of action of Platycodon grandiflorum's "drug-inducing" effect. A correct and objective understanding of the efficacy of this "drug-inducing" effect will help better guide clinical practice. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 This is the "drug-active ingredient-disease target" network of Platycodon grandiflorum. DETAILED DESCRIPTION

[0035] The present invention will be further described in detail below through specific embodiments in conjunction with the accompanying drawings.

[0036] Example 1

[0037] 1.1 Database and Software

[0038] TCMSP database (https: / / old.tcmsp-e.com / tcmsp.php);

[0039] Uniprot database (https: / / www.uniprot.org / );

[0040] SWISSTargetPrediction(http: / / swisstargetprediction.ch / );

[0041] DisGeNET database (https: / / www.disgenet.org / ) and GeneCards database (https: / / www.genecards.org / );

[0042] STRING database (https: / / cn.string-db.org / );

[0043] Software: Cytoscape 3.10.2.

[0044] 1.2 Collection and screening of chemical components from Platycodon grandiflorum

[0045] All chemical components related to the search for Platycodon grandiflorum as a keyword were retrieved through the Chinese medicine pharmacology system database and analysis platform, and related literature was searched for supplementary explanations. The conditions were set to oral availability OB ≥ 10% and DL ≥ 0.18 (in order to obtain more and more effective Platycodon grandiflorum active ingredients), and Platycodon grandiflorum was screened to identify the main effective compounds and obtain chemical components that met the conditions.

[0046] 1.3 Obtaining target genes corresponding to effective compounds of Platycodon grandiflorum

[0047] Import the SMILE name (in Pubchem) of the effective compounds of Platycodon grandiflorum into the SWISSTargetPrediction database to obtain the gene name of a single main component (the common name in this figure is the gene name, which is also the protein name. The protein name can be directly used to construct the PPI network); import the target name of the main component in the figure into Uniport (http: / / www.uniprot.org / ) to obtain the gene name. One target corresponds to one gene name. The gene name is obtained in Uniport and saved in Excel. Each main component must be corresponding and saved, and the five first located humans (Homo) are standardized and converted to obtain the target gene name. After the converted target gene names are integrated, the duplicate values are deleted.

[0048] 1.4 Disease gene acquisition

[0049] The DisGeNET database and GeneCards database are two disease gene data analysis platforms. First, "pneumonia" and "sore throat" were used as keywords to search the above two databases. Then, the results obtained from the disease genes in the above gene databases were merged and removed, and the known targets for lung abscess and sore throat were obtained. The intersection targets of Platycodon grandiflorum for treating pneumonia and sore throat were analyzed. By removing duplications, the randomness was relatively avoided to a certain extent.

[0050] 1.5 Establishing a common target for Platycodon grandiflorum, lung abscess, and sore throat

[0051] The intersection of Platycodon grandiflorum and lung abscess and sore throat disease targets was calculated and constructed by Venny 2.1.0 to obtain the common targets of Platycodon grandiflorum and lung abscess and sore throat diseases, and the results were displayed in a Venny diagram.

[0052] 1.6 Constructing a drug-active ingredient-disease target network diagram

[0053] A drug-disease-compound-target network was constructed using Cytoscape v3.10.2 software. The compiled database was imported into Cytoscape v3.10.2. Valid gene targets were searched and ineffective ones deleted. The graph was then reshaped into a circular shape using the Layout option. The appropriate shape, color, width, and height for the compounds and genes were selected in the Style option. Nodes represented the components of Platycodon grandiflorum and their targets, while edges represented the interactions between the active drug ingredients and the disease targets. This completed the drug-active ingredient-disease target network.

[0054] 1.7 Construction of a protein-protein interaction network (PPI) for common targets of Platycodon grandiflorum and lung abscess and sore throat

[0055] The target proteins of the common targets identified for Platycodon grandiflorum, lung abscess, and sore throat were imported into the String database. Multiple proteins, Organism, and Homo sapiens were selected, free nodes were hidden, and all other parameters were set to default values. This completed the protein-protein interaction network diagram between Platycodon grandiflorum and the common targets.

[0056] 1.8GO functional enrichment analysis and KEGG pathway enrichment analysis

[0057] The intersection data of lung abscess and sore throat disease targets were uploaded to the David platform, the species was selected as "Homosapiens", and its biological processes and pathways were analyzed. The obtained data was imported into the microbial information platform and then visualized to obtain a bubble chart.

[0058] result

[0059] 2.1 Screening of the main active ingredients in Platycodon grandiflorum

[0060] A search using the keyword "Platycodon grandiflorum" in the TCMSCP database yielded 130 active ingredients. Setting the screening value of OB ≥ 10% yielded 52 active ingredients. After selecting active ingredients with a DL ≥ 0.18, 22 ingredients met both screening values, including trochol, acacetin, and spinasterol. See Tables 1 and 2.

[0061] Table 1

[0062]

[0063] Table 2

[0064]

[0065] 2.2 Targets of Platycodon grandiflorum ingredients

[0066] 22 active ingredients were entered into Pubchem, ultimately obtaining gene targets for seven active ingredients. No targets or sequences were obtained for the remaining ingredients. After screening, these were sequentially imported into the SWISSTargetPrediction database (http: / / old.swisstargetprediction.ch / ) to obtain the gene names of the individual main ingredients. Each main ingredient was mapped and saved, and the five first located in human (Homo) were standardized to obtain target gene names. After the converted target gene names were integrated, duplicate values were deleted.

[0067] 2.3 Collection of disease targets related to lung abscess and sore throat

[0068] Using the search keywords "pneumonia" and "sore throat" in the disease database DisGeNET and GeneCards, relevant targets were summarized and the data was deduplicated. Lung abscess and sore throat disease targets were analyzed using the Venny online platform, resulting in 101 and 104 overlapping targets, respectively, indicating potential targets for Platycodon grandiflorum in treating lung abscess and sore throat.

[0069] 2.4 Construction and analysis of the “drug-active ingredient-disease target” network

[0070] The above-mentioned drug and disease intersection targets were imported into Cytoscape_v3.10.2 software to construct a "drug-active ingredient-disease target" network diagram. In the diagram, the drug Platycodon grandiflorum is represented in red, the active ingredient of the drug is represented in yellow, and the target shared by the drugs is represented in blue. Figure 1 As shown in the figure, there are 7 active ingredients in Platycodon grandiflorum and 104 common drug targets.

[0071] 2.5 Construction of PPIs with Platycodon grandiflorum as a common target for lung abscess and sore throat

[0072] Import the intersecting targets of Platycodon grandiflorum and the diseases into the String database. Set the "minimum required interaction score" value to "high confidence (0.700)", hide the free targets, set the Clustering Options "k-means clustering" and "number of clusters" to 4, export the .txt file, and then generate a protein-protein interaction network diagram. The core targets of the intersecting targets of Platycodon grandiflorum and sore throat diseases include EGFR, MMP9, NR3C1, IGF1R, and MAPK14. The core targets of the intersecting targets of Platycodon grandiflorum and lung abscess diseases include EGFR, MAPK3, MAPK14, NR3C1, PTGS2, and ESR1.

[0073] 2.6 GO and KEGG pathway enrichment analysis of the intersection targets between Platycodon grandiflorum and lung abscess and sore throat

[0074] Signal pathway analysis of the drug Platycodon grandiflorum for the treatment of lung abscess and sore throat was performed using the David database. After visualization, the intersection targets of the GO map and the KEGG analysis map were obtained. The processed intersection targets were involved in biological processes (BP), cellular components (CC), and major molecular functions (MF) and were saved. The KEGG pathway enrichment results were then summarized according to the KEGG classification to obtain the following result diagram. GO biological process enrichment analysis and KEGG pathway enrichment analysis showed that lung abscess and sore throat have similar etiologies and pathogenesis. The top 10 pathways were the PI3K-Akt signaling pathway, the C-type lectin signaling pathway, the PD-1 and PD-L1 signaling pathways, and the chemical carcinogenic receptor activation signaling pathway.

[0075] The above are merely preferred embodiments of the present invention and are not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention shall be included within the scope of protection of the present invention.

Claims

1. A method for exploring the upward movement of Platycodon grandiflorum based on network pharmacology, characterized by: The following steps are involved: Step 1: Collection and screening of chemical components from Platycodon grandiflorum Platycodon grandiflorum is screened to find effective compounds that meet the conditions and obtain chemical components that meet the conditions; Step 2: Obtaining target genes corresponding to effective compounds of Platycodon grandiflorum Through the effective compounds of Platycodon grandiflorum, the corresponding gene names are obtained, and one target corresponds to one gene name; Step 3: Obtaining disease genes By using a disease associated with Platycodon grandiflorum as a keyword search to obtain corresponding known targets, the corresponding targets are analyzed; Step 4: Constructing common disease targets Construct the intersection of Platycodon grandiflorum and disease targets to obtain the common targets of Platycodon grandiflorum and diseases; Step 5: Construct a network diagram of drugs, active ingredients, and disease targets By searching for effective gene targets, a network diagram of drugs, active ingredients and disease targets is constructed; Step 6: Constructing a network diagram of protein-protein interactions between Platycodon grandiflorus and common targets. By importing the target proteins of the common targets obtained by screening into the database, a network diagram of protein-protein interactions between Platycodon grandiflorus and common targets is constructed. Step 7: GO functional enrichment analysis and KEGG pathway enrichment analysis The disease target intersection data was uploaded to the David platform, and the data was visualized to obtain a bubble chart. Then, based on the bubble chart and combined with the existing information, the mechanism of Platycodon grandiflorum leading the drug upward was analyzed.

2. A method for exploring the upward flow of Platycodon grandiflorum based on network pharmacology according to claim 1, characterized in that: Platycodon grandiflorum was screened, and 130 effective compounds were screened out, of which 22 met the requirements.

3. A method for exploring the upward flow of Platycodon grandiflorum based on network pharmacology according to claim 2, characterized in that: Based on the 22 active ingredients, 7 target genes corresponding to 7 effective compounds of Platycodon grandiflorum were obtained.

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