Mechanism of Shenfu Decoction in the treatment of critically ill patients with Coronavirus Disease 2019 (COVID-19) based on network pharmacology

Objective:The purpose of this thesis is to explore the mechanism of ShenFu Decoction in the treatment of critically ill patients with COVID-19 based on network pharmacology.Methods:The primary active ingredients and potential targets of ShenFu Decoction were searched from the TCMSP database.The targets of COVID-19 were obtained by searching the GeneCards and OMIM databases.A ShenFu Decoction-compound-target-COVID19 network and a protein-protein interaction(PPI)network were respectively constructed through the Cytoscape 3.5.1 software and the STRING database.Gene Ontology(GO)function enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis were performed via Bioconductor bioinformatics software package and R programming language.Results:ShenFu Decoction contains 255 compounds and 94 potential targets.43 primary active ingredients were searched from the TCMSP database with oral bioavailability(OB)≥30%and drug-likeness(DL)≥0.18 as the retrieval condition.Numbers of targets of COVID-19 were 352 by searching the GeneCards and the OMIM databases.16 key targets were acquired by intersecting the targets of drug with the targets of disease.There were 49 GO terms and 102 pathways after analyzing GO and KEGG.Conclusion:Kaempferol,ginsenoside rh2,beta-sitosterol,Stigmasterol and Deoxy andrographolide might be the main active ingredients which may cause the inhibition of the SARS-CoV-23CL hydrolase activity and regulate ACE2.As a result,the antiviral effect,immunoregulation,targeting cytokine storm of SFD may play an important role in the treatment of critically ill patients with COVID-19 through regulating multiple signaling pathways such as AGE-RAGE signaling pathway in diabetic complications,IL-17 signaling pathway,C-type lectin receptor signaling pathway,HIF-1 signaling pathway.

Network pharmacological study on the mechanism of shenfu decoction in the treatment of cardiotoxicity by doxorubicin

Objective:Doxorubicin is an efficient anthracycline drug for the treatment of tumor,however,its cardiotoxicity restricts the clinical application.Shenfu decoction has good clinical effect,but the pharmacological mechanism is not fully clarified.Method:The active components and potential targets of shenfu decoction were screened by TCMSP database,disease targets of doxorubicin-induced cardiotoxicity were collected by Genecards and OMIM database,and the network diagram of"drug-components-target-disease"was constructed by Cytoscape software.PPI network was constructed by STRING database.The target of action of the drug and the disease gene were mapped for GO and KEGG signal pathway analysis.Results:The study found that there are 52 main effective components of shenfu decoction,and 76 genes are involved in the potential therapeutic targets,among which 24 genes are potential targets of shenfu decoction in the treatment of doxorubicin-induced cardiotoxicity.The protein interaction network suggested that BCL2、BAX、CASP9、CASP3、MAPK8 may be the core target.GO enrichment analysis showed 52 cellular biological processes,and enrichment analysis of KEGG pathway revealed 99 involved signaling pathways,including TNF,apoptosis signaling pathways,etc.Conclusion:In this study,the network of"drug-components-target-disease"was constructed through network pharmacology,and it was found that the mechanism of"shenfu decoction"in the treatment of doxorubicin-induced cardiotoxicity involves multiple targets and pathways,which is conducive to guiding clinical medication.

Cardioprotection of Shenfu preparata on cardiac myocytes through cytochrome P450 2J3

To evaluate whether Shenfu injection （SFI） protects against cardiac myocyte injury induced by Fupian injection （FPI） in vitro. METHODS： H9c2 cells were separately treated with FPI, Renshen injection （RSI） and SFI. Cell viability, lactate dehydrogenase （LDH） release, spontaneous beating rate of primative cardical cells, caspase-3/7 activity, cell apoptosis, and cytochrome P450 2J3 （CYP2J3） mRNA expression were analyzed. RESULTS： The viability of H9c2 cells treated with SFI （37 and 75 mg/mL） was significantly higher than that of H9c2 cells treated with FPI （25 and 50 mg/mL） （P〈0.05, P〈0.01, respectively）. LDH activity of H9c2 cells treated with SFI （75 mg/mL） was significantly decreased （P〈0.01） compared with that of H9c2 cells treated with FPI （50 mg/mL）. SFI （150 mg/mL） significantly attenuated FPI （100 mg/mL）-induced spontaneous beating rate decrease in primary myocardial cells after 4-hour treatment. Compared with FPI （12 and 25 mg/mL）, SFI （18 and 37 mg/mL） treatment could effectively reverse the change of caspase-3/7 activity （P〈0.01 and P〈0.01, respectively）. Compared with FPI （6 and 25 mg/mL）, apoptotic cells decreased significantly （P〈0.05, P〈0.01, respectively） when H9c2 cells were incubated with SFI （9 and 37 mg/mL）. The expression of CYP2J3 mRNA was down-regulated by FPI, while RSI and SFI could up-regulate the expression of CYP2J3 （P〈0.01）, which suggested the potential mechanism of protection of RSI against cardiac myocyte damage induced by FPI treatment. CONCLUSION： These observations indicate that SFI has the potential to exert cardioprotective effects against FPI toxicity. The effect was possibly correlated with the activation of CYP2J3.

Revealing the synergistic mechanism of Shenfu Decoction for anti-heart failure through network pharmacology strategy

The present study was designed to investigate the targets and synergistic mechanism of Shenfu Decoction(SFD) in the treatment of heart failure.A heart failure animal models was established to evaluate the pharmacological effects of SFD for anti-heart failure, then constructed ingredient-target interaction network by developing ingredient and target databases, the Discovery sdudio software was used for molecular docking.In addition, we validated the predicted protein targets of active ingredients in SFD by using surface plasmon resonance(SPR) technology.Our results demonstrated that SFD could enhance ejection fraction, alleviate myocardial histopathological characteristics, and reduce the level of angiotensin converting enzyme(ACE), aldosterone(ALD), atrial natriuretic polypeptide(ANP) and Renin(REN) in heart failure rat model.In addition, the ingredient database including 349 constituents and target database including 236 proteins were established, and 75 proteins were screened and identified by molecular docking strategy.22 core target proteins were identified through network pharmacology, and the component-core target network was constructed.Finally,the affinity between the compounds and targets were verified by the SPR analysis method.The present study suggested that SFD may act on ACE 2, REN, ACE, ICAM-1, EGF, HTR2B, PARP1, NPPB and other proteins through AC, BAC, ACN, Re, Rg1, Rb1 to exert synergistic effects against heart failure.

Efficacy of Shenfu decoction on sepsis in rats with condition induced by cecal ligation and puncture

OBJECTIVE:To evaluate the efficacy of Shenfu decoction(SFD)prepared with a traditional Chinese formula,on sepsis in rats with the condition induced by cecal ligation and puncture(CLP),and to study the possible mechanism underlying its action.METHODS:Forty clean-grade male Sprague-Dawley rats were divided randomly into three groups:normal control group(NCG,n=10),model control group(MCG,n=15)and Shenfu decoction group(SFDG,n=15).Sham-operated rats in NCG were served as operation control,while rats in both MCG and SFDG were exposed to CLP,a procedure to develop experimental sepsis.Rats in SFDG were administered with SFD by gavage(3 mg/g of body weight,twice a day)2 h prior to CLP and directly after successful CLP,while rats in NCG and MCG were gavaged with equivalent volume of sterilized water.Rats in all groups were starved with free access to drink.After 24 h of administration,the mortality of rats in each group was assessed.The indicators of inflammatory response[the peritoneal inflammation by Simon's method Classification as well as serum concentrations of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)by enzyme linked immunosorbent assay(ELISA)]in survival rats were evaluated.The indicators of gut barrier[The intestinal mucosal injury index,serum concentrations of D-lactic acid and secretory IgA(sIgA)in intestinal mucosa by ELISA,as well as gut microbiota by16S rRNA gene sequencing]in survival rats were evaluated.RESULTS:The mortality(20%)of rats in SFDG was lower than that(33.3%)of the MCG(P<0.01).The mortality(20%)of rats in SFDG was lower than that(33.3%)of the MCG(χ2=6.533,P=0.011).Compared with the MCG,the peritoneal inflammation as well as serum concentrations of TNF-αand IL-6 decreased significantly in SFDG(all P<0.01).Compared with the MCG,the IMII,serum concentrations of D-lactic acid,sIgA in intestinal mucosa were alleviated by SFD treatment(all P<0.01).Increase in levels of Proteobacteria and reduction levels of Bacteroidetes induced by sepsis were observed,and these two disturbed gut microbiota phyla could be regulated after SFD treatment.Increase in levels of Proteobacteria and reduction levels of Bacteroidetes induced by sepsis were observed,and these two disturbed gut microbiota phyla could be regulated after SFD treatment.CONCLUSION:SFD may play a protective role in sepsis by alleviating sepsis-induced inflammatory response and gut barrier damage in rats.

EXPERIMENGAL WORK AND RESEARCH Experimental Study on Protective Effect of Chinese Herbal Medicine on Glucocorticoid Receptor

Objective: To probe the relationship of glucocorticoid receptor and some Chinese medicinal herbs. Methods: The model of Qi-Yang exhaustion and Qi-Yin exhaustion were made with hemorrhagic rats and heat-stressed rats respectively. The effect of Shenfu decoction (SFD) and Shengmai powder (SMP) on plasma glucocorticoid (GC) and its receptor (GcR) in hepatic cytosol of the models were measured respectively. Results: The activity of GcR decreased in both models, while their blood level of GC increased markedly. SFD and SMP showed no regulating effect on blood GC, but displayed obvious up-regulation on GcR level in both models. Conclusion: SFD and SMP could up-regulate the activity of GcR in Qi-Yang and Qi-Ying exhaustion models.