The potential mechanism of prevention and treatment of COVID-19 by shufeng jiedu capsule was studied based on network pharmacology

Jing-Wen Huang, Li-Feng An, Xue Han, Qing-Wei Li, Hui Xue, Liu Yang, Jian-Fei Duan

Jiamusi College, Heilongjiang University of Traditional Chinese Medicine, Jiamusi 150040, China

Keywords:Shufeng Jiedu Capsule Corona Virus Disease 2019 (COVID-19)Network pharmacology Target Signaling pathway

ABSTRACT Objective: To investigate the mechanism of Shufeng Jiedu Capsule(SJC) for treatment of COVID-19 based on network pharmacology. Methods: Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) was used to obtain the active components and targets information of Polygoni Cuspidati Rhizoma Et Radix, Forsythiae Fructus, Istidis Radix, Radix Bupleuri, Herba Patriniae, Verbenae Herb, Phragmitis Rhizoma and licorice in SJC. The Genecards databases were used to obtain COVID-19 targets. The meridian tropisms of each herb in SJC were collected from ETCM Database.the proteins interations network were build by STRING database. The GO and KEGG pathways were analyzed by the computer R language. Results: SFC contains 8herbs, 176 compounds and the corresponding targets 237. 48 COVID-19 targets are treated by SJC. Such as IL-10, IL-6, PTGS1, PTGS2, GSK3B, STAT-1, IL-17 signaling pathway, HIF-1 signaling pathway, Toll-like receptor signaling pathway, T cell receptor signaling pathway and VEGF signaling pathway may be potential targets and signaling pathways for the prevention and treatment of COVID-19. Conclusion: The COVID-19 cantreatmented the potential targets and signaling pathways by the SJC, and play the role of antipyretic, anti-inflammatory, balance immunity, antiviral and so on. It will provide strong support for the later stage experiment and clinical application of SJC.

1. Introduction

The novel coronavirus pneumonia outbreak at the end of 2019, referred to as the COVID-19 [1], has a long incubation period, is highly infectious and generally susceptible, and has a great impact on people's health, economic development and social stability. (COVID-19) At present, how to prevent early and effectively is the top priority. It is found that the early intervention of traditional Chinese medicine can effectively reduce symptoms, block the course of disease, shorten the treatment cycle, and play an important role in the comprehensive advantages of multi-component, multi-target, broad-spectrum antibacterial, antiviral and balanced immunity [2]. In the latest novel coronavirus pneumonia diagnosis and treatment plan (trial version 7) issued by the National Health Protection Committee, the importance of TCM diagnosis and treatment is also emphasized. It is pointed out that during the medical observation period, if the clinical manifestations are fatigue and fever, the Chinese medicine such as Shufeng Jiedu Capsule (granule) and so on should be recommended.

Shufeng Jiedu Capsule (SFC)[3], formerly known as "Qudu powder", is a classic prescription that has been circulating for a hundred years. It is mainly used to treat cold, diphtheria, mumps, tonsillitis and other diseases. The traditional Chinese medicine was widely used after the removal of Cynanchum wilfordii and the addition of Istidis Radix, Radix Bupleuri and Phragmitis Rhizoma. It is a compound preparation composed of 8 traditional Chinese medicines, Polygoni Cuspidati Rhizoma et Radix, Forsythiae Fructus, Istidis Radix, Radix Bupleuri, Herba patriniae, Verbenae Herb, Phragmitis Rhizoma and licorice. It has obvious inhibitory effect on influenza virus through modern pharmacological research and has been widely used in many fields The diagnosis and treatment plan is listed as the first choice Chinese patent medicine for the treatment of influenza.

Network pharmacology is a research method based on systems biology and multi-directional pharmacology [6]. It is especially suitable for studying the action relationship of multi-components and multi-targets of traditional Chinese medicine, and is helpful to show the action mechanism of traditional Chinese medicine components [7-9]. Therefore, this study uses the technical means of network pharmacology to study the action target and signal pathway of SFC and analyze its potential molecular mechanism for the prevention and treatment of COVID-19, in order to provide strong support for the later experiment and clinical application of SFC.

2.Materials and methods

2.1Screening the target compounds and targets of SFC

Input the pinyin name of traditional Chinese medicine in SFC into the pharmacological analysis platform TCMSP database of traditional Chinese medicine system (http://tcmspw.com/tcmsp.php) , set DL ≥ 0.18line OB ≥ 30%, and obtain the qualified active components and corresponding target information of SFC. Then the network pharmacology related research was carried out through the Uniport database (https://www.uniprot.org/) query standard protein target ID.

2.2Construction of drug-meridian network diagram of SFC

The meridian tropism of each drug in SFC was found in ETCM database (http:www.nrc.ac.cn:9090/ETCM/index.php/Home/Index/), and then the drug-meridian network diagram was drawn by Cytoscape 3.6.1 software, in order to directly understand the attribution and action orientation of viscera organs and meridians of SFC.

2.3Draw the PPI network diagram of protein-protein interaction of SFC in the prevention and treatment of COVID-19

By using Genecards database, taking "novel coronavirus pneumonia" and "coroavirus disease 2019" as keywords and selecting species as "Homosapiens", the target information related to COVID-19 was intersected with the action target of SFC, so as to obtain the potential target of SFC in the prevention and treatment of COVID-19. Import the data into the STRING database (https://string-db.org/) , and the species selection "HomoSapiens" and "highestconfidence (0.400)" were set to draw the PPI network diagram of protein-protein interaction of SFC in the prevention and treatment of COVID-19.

2.4Functional enrichment of GO and Analysis of KEGG Pathway

The target information of SFC in preventing and treating COVID-19 was calculated by computer programming, and the enrichment of GO biological process and KEGG pathway was analyzed by using "Clusterprofiler" in computer R language and setting P ＜ 0.01.

3. Result

3.1Chemical constituents and targets of SFC

The effective components and targets of the drugs in the prescription of SFC were obtained through TCMSP database, and the compounds in Polygoni Cuspidati Rhizoma et Radix, Forsythiae Fructus, Radix Bupleuri, Herba patriniae, Verbenae Herb , Phragmitis Rhizoma, licorice and Istidis Radix were retrieved, and a total of 176 compounds were screened by OB ≥ 30%,DL ≥ 0.18, Polygoni Cuspidati Rhizoma et Radix has 8 compounds, Forsythiae Fructus has 18 compounds, Radix Bupleur has 12 compounds, Herba patriniae has 11 compounds, Verbenae Herb has 11 compounds, Phragmitis Rhizoma has 1 compounds, licorice has 84 compounds and 31 Phragmitis Rhizoma has 31 compounds. As Shown in Table 1. At the same time, the action targets corresponding to each chemical composition were obtained, and the repetitions were deleted, and a total of 237 protein targets of SFC were obtained.

Table 1. The qualified chemical components and the number of corresponding targets in SFC.

3.2Analysis of drug-meridian network diagram of SFC

The meridian tropism of 8 traditional Chinese medicines in SFC was obtained by ETCM database, and then the drug-meridian network diagram was drawn by Cytoscape3.6.1 software. it can be seen from figure 1 that the connection degree of lung meridian is the largest, and that of lung meridian, liver meridian and stomach meridian are 5, 4 and 4 respectively, indicating that SFC mainly belongs to lung meridian, followed by liver meridian and stomach meridian. It is suggested that SFC has a good preventive and therapeutic effect on respiratory and lung diseases caused by external evil, and can regulate both liver and stomach meridians.

Figure 1. meridian network of drugs in SFC(red represents medicine, green represents the viscera, meridians and collaterals to which it belongs)

3.3Analysis of PPI Network Diagram of protein-protein interaction in SFC

The protein target of SFC was intersected with the associated target of COVID-19. After removing the repetition, 48 targets were obtained, which were introduced into STRING, and finally the protein-protein interaction PPI network map of SFC for prevention and treatment of COVID-19 was obtained. As Shown in Figure 2. The targets of IL-10, IL-6, PTGS1, PTGS2, GSK3B and STAT-1 in the protein interaction are closely related to antipyretic, antiinflammatory and immune regulation, which can be used as the potential core targets of SFC in the prevention and treatment of COVID-19.

Figure 2. protein-protein interaction network of SFC in prevention and treatment of COVID-19

3.4 GO enrichment analysis and KEGG pathway enrichment analysis

A total of 80 items were obtained by GO biological process enrichment analysis, and the top 20 items were selected. As Shown in Figure 2. Including cytokine receptor binding, cytokine activity, phosphatase binding, chemokine receptor binding, heme binding, growth factor receptor binding and so on, involving cell regulation, biological regulation, immunity, metabolism and so on. It is suggested that the effective components of SFC can control COVID-19 by regulating a variety of biological ways.

Figure 3. GO bubble chart of SFC in the prevention and treatment of COVID-19

Figure 4. Network diagram of target- signaling pathway for prevention and treatment of COVID-19 with SFC Discuss

A total of 157 signal pathways were obtained by KEGG enrichment analysis. According to the information obtained, the target-signaling pathway network diagram was drawn. As Shown in Figure 2. Among them, Hepatitis B, Kaposi sarcoma-associated herpesvirus infection, Human cytomegalovirus infection, Hepatitis C, Non-small cell lung cancer, Tuberculosis, Influenza A and so on are related to virus infection. The signal pathways closely related to antipyretic, antiinflammatory, antiviral and immune regulation are IL-17 signaling pathway, HIF-1 signaling pathway, Toll-like receptor signaling pathway, T cell receptor signaling pathway and VEGF signaling pathway , which can be used as potential signal pathways to study the prevention and treatment of COVID-19 by SFC.Novel coronavirus infected the human body, invaded the respiratory system, there is a certain incubation period, when the human immunity is low, unable to remove the virus, and then cause a series of inflammatory reactions, patients with cough, sore throat, expectoration, wheezing and other clinical manifestations. At the same time, according to the region, climate, physique, condition, patients are also accompanied by different concurrent diseases, such as fatigue, fever or gastrointestinal discomfort, can flexibly take traditional Chinese medicine compound or proprietary Chinese medicine for symptomatic treatment. SFC is a commonly used traditional Chinese medicine preparation for the treatment of influenza [10], with broad-spectrum antiviral effect [11]. Polygoni Cuspidati Rhizoma Et Radix as the sovereign medicine, bitter slightly astringent slightly cold, dispelling wind and dampness, clearing heat and detoxification, relieving cough and resolving phlegm; minister to clear heat and detoxification, dissipate Forsythiae Fructus, Istidis Radix; It is assistanted with Radix Bupleuri for dispeling expathogens to relieving fever, clearing heat and removing toxins, draining pus and relieving blood stasis, Herba Patriniae, and Verbenae Herb for promoting blood circulation and inducing diuresis, and Phragmitis Rhizoma for clearing heat and promoting fluid. Licorice is used to reconcile various medicines. All kinds of medicines are used together to play the function of dispelling wind and clearing heat, detoxification and pharynx[12]. During COVID-19 's medical observation period, it is recommended for people with fatigue and fever.

In this paper, the method of network pharmacology was used to analyze SFC from the molecular level. 152 chemical constituents and 214 target information were selected from 8 kinds of traditional Chinese medicine, and the targets such as IL-10, IL-6, PTGS1, PTGS2, GSK3B, STAT-1,and the signaling pathway such as IL-17 signaling pathway, HIF-1 signaling pathway, Toll-like receptor signaling pathway, T cell receptor signaling pathway and VEGF signaling pathway were screened. It is closely related to antipyretic, anti-inflammatory, anti-virus, regulation of apoptosis, enhancement of body defense system and regulation of immune function. It is speculated that SFC may have preventive and auxiliary therapeutic effects on COVID-19.

Among the above targets analyzed by network pharmacology, proinflammatory factor IL-6 plays an important role in the process of immunity and inflammation [13], especially in the late stage of inflammation. IL-10 is an anti-inflammatory factor, which can alleviate the inflammatory response caused by coronavirus by inhibiting inflammatory factors and regulating immune function [14]. PTGS (prostaglandin-endoperoxide synthase) is not only the target of non-steroidal anti-inflammatory drugs, but also the key enzyme of prostaglandin synthesis, which mediates fever and increases body temperature by promoting the synthesis of prostaglandin E2[15]. It mainly includes two subtypes: PTGS1 and PTGS2. PTGS1 can promote vasoconstriction and proliferation of vascular smooth muscle cells [16], PTGS2 [17] can induce inflammatory pathological reaction of redness, swelling, heat and pain [18]. Antipyretic and antiinflammatory effects can be achieved by inhibiting the activities of PTGS1 and PTGS2. GSK3B (glycogen synthase-3) can regulate the proliferation, activation and expression of inflammatory factors of T cells [19]. STAT-1 (transcriptional activator 1) is an important member of the STAT family, and its signal transduction pathway is closely related to cell proliferation and apoptosis[20]. STAT1 [21] is also an important molecule in the inflammatory signaling pathway. Under the condition of novel coronavirus infection, the regulation of STAT1 may control the occurrence of inflammation and fever. Therefore, SFC may play a variety of pharmacological effects, such as antipyretic, anti-inflammatory, regulating immune function and so on, aiming at COVID-19 's fatigue with fever during the medical observation period by regulating the above key targets.

Among the five signaling pathways enriched by KEGG, the pathway related to immune response is the signal pathway of T cell receptor signaling pathway . T cells are the core effector molecules of adaptive immune response, which can effectively eliminate invasive pathogens and infected cells, and play a key role in adaptive immune response against infection [22]. The pathways related to antiinflammatory and immune regulation are IL-17 signaling pathway and HIF-1 signaling pathway : IL-17 signaling pathway[23] can regulate the immune system indirectly by regulating the production and release of chemokines, pro-inflammatory cytokines and antimicrobial peptides, and play an important role in the pathological process of various inflammatory reactions and autoimmune diseases. HIF-1 signaling pathway plays an important role when the body is in a state of hypoxia. Novel coronavirus is easy to invade the lungs through the respiratory tract, causing hypoxia and inflammation in the lungs [24]. Regulating HIF-1 signaling pathway can regulate immune function, eliminate inflammatory reaction, induce neovascularization [25-26], and help to relieve lung symptoms such as cough, expectoration and wheezing. The pathway related to antipyretic, immunomodulatory and antiviral is the Toll-like receptor signaling pathway [27]. Toll-like receptor is the first barrier against pathogen invasion and clearance of its own waste, and plays an important role in the process of immune regulation. In addition, it also has antipyretic and antiviral effects [28].

To sum up, SFC can prevent and assist the treatment of COVID-19 by means of antipyretic, anti-inflammatory, antiviral and regulating immune function with the help of the above targets and signaling pathways, and explain its potential mechanism from the molecular level. it corresponds to the efficacy of SFC in dispelling wind and clearing heat, detoxification and pharynx. Due to the limitations of network pharmacology itself, the database can not contain all the components and target information, and there may be deviations, which will be verified by animal experiments in the future.