Screening Of Antiviral Medicinal Plants And Preliminary Study Of The Mechanism Of Formula SWJTU-MY20

The viruses that cause deadly infectious diseases such as influenza,AIDS and Ebola haemorrhagic fever endanger human health all the time.The outbreak of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infection in 2019 caused huge losses worldwide,and so far,there is no drug with specific effect on SARS-CoV-2.Due to the limitations of the understanding of the virus,the research and development of related antiviral drugs is limited.Plant medicine contains a variety of bioactive substances such as polysaccharides and alkaloids,which can exert the efficacy that synthetic substances are difficult to achieve,and has the advantages of low toxicity and side effects,wide antiviral spectrum,no environmental pollution,and contains natural minerals,vitamins,nutrients,etc.,with treatment,prevention,nutrition and other effects.China has a long history and rich experience in the treatment of viral diseases with plant medicine.Therefore,it has become an important way to find safe,effective and antiviral drugs from medicinal materials.In this study,an effective antiviral compound of plant medicine was obtained by screening plant medicinal materials.After review of a large number of books and documents,254 kinds of medicinal materials were summarized.Using M13 phage and adenovirus as model virus,the antiviral effect of alcohol extract of medicinal materials was determined in vitro,and the medicinal materials with good antiviral effect were obtained,such as Phyllanthi Fructus,Phyllanthus urinaria L.,Inula lineariifolia Turcz.and so on.The prescription of compatibility was selected to determine the optimized prescription and named as formula SWJTU-MY20.It was found that formula SWJTU-MY20 had strong inhibitory effect on M13 bacteriophage and adenovirus.SWJTU-MY20 has strong inhibitory effect on several common biological pathogens at the same time: the MIC and MBC of Candida albicans,clinical Staphylococcus aureus,Methicillin-resistant Staphylococcus aureus(MRSA),clinical Escherichia coli and Pseudomonas aeruginosa are 7.0% and 8.0%,2.0% and 3.0%,2.0% and 3.0%,9.0% and10.0%,5.0% and 6.0%,respectively.It is revealed that formula SWJTU-MY20 has broadspectrum antiviral and bactericidal effects.In this study,the active components in formula SWJTU-MY20 were collected and screened with the help of multiple databases,its action targets were found and predicted,and the formula-gene-disease network was constructed.Quercetin,luteolin,apigenin,ursolic acid,baicalein,kaempferol,myricetin and emodin were quickly obtained from this network.Among them,quercetin,apigenin,kaempferol,luteolin,wogonin,myricetin,can be converted into flavonoids,with a wide range of antiviral activity.In the PPI interaction analysis of formula potential targets,the top ten targets with Degree value are STAT3,JUN,TNF,RELA,IL6,AKT1,MAPK1,FOS,IL1 B and CXCL8,they are speculated to be the core targets in the formula intervention disease network.The core target also contains a variety of cytokines or chemokines closely related to viral infection: TNF,IL6,IL1 B,CXCL8,speculate that the formula may affect the inflammatory response caused by viral infection and play a therapeutic role.In the subsequent GO and KEGG enrichment analysis,it was found that the biological function was mainly enriched in the processes related to cytokines and inflammation,and many signaling pathways related to viral infection were significantly enriched,such as human cytomegalovirus infection,Kaposi sarcoma-associated herpesvirus infection,hepatitis C,hepatitis B,influenza A,EBvirus infection,T-lymphocytic leukemia virus infection.It is speculated that SWJTU-MY20 can play a therapeutic role by regulating some inflammatory responses to these viral infection pathways.Further molecular simulation and docking of SWJTU-MY20 anti-SARS-CoV-2 novel coronavirus showed that the eight key components of formula SWJTU-MY20 combined well with three key targets of novel coronavirus: Mpro,ACE2 and RBD protein of S-prickle glycoprotein,which revealed that the formula may play a possible role in anti-novel coronavirus.Finally,some related medicinal materials in SWJTU-MY20 were identified by DNA bar code using ITS2 and psbA-trnH molecular fingerprinting technique.Firstly,the genomic DNA,was extracted from the medicinal materials,and then the ITS2 sequence and psbAtrnH sequence were amplified by DNA,and the ITS2 and psbA-trnH sequences were sequenced.The sequences of ITS2 and psbA-trnH were searched in NCBI database,the sequences of related species were downloaded,and the phylogenetic tree was constructed for phylogenetic analysis.Five kinds of medicinal materials in formula SWJTU-MY20 were identified by ITS2 bar code: Isatidis Radix,Kummerowia striata(Thunb.)Schindl.,Trollius chinensis Bunge,Carthamus tinctorius L.,Phyllanthus urinaria L.,and five kinds of medicinal materials in formula SWJTU-MY20 were identified by psbA-trnH bar code:Isatidis Radix,Kummerowia striata(Thunb.)Schindl.,PLatycladus orientalis(L.)Franco,Trollius chinensis Bunge and Carthamus tinctorius L..According to the results of ITS2 and psbA-trnH bar code identification,four kinds of medicinal materials were identified in the formula: Radix Isatidis,Cornus angustifolia,Golden Lotus and Carthamus tinctorius.To sum up,this study successfully developed a plant drug formula SWJTU-MY20 with antiviral and broad-spectrum antibacterial activity through high-throughput screening of ethanol extracts from medicinal plants.The antiviral mechanism of formula SWJTU-MY20 was studied by network pharmacology combined with molecular docking.It provides an important basis for the research and development of antiviral plant medicine formula.