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Objective:To reveal the effective components， targets and possible mechanisms of Qinggan Huayu granules in the treatment of non-alcoholic fatty liver disease （NAFLD） and liver cancer based on network pharmacology and experimental verification， and to provide a basis for its rational interpretation of treating different diseases with same method for NAFLD and liver cancer. Method:Based on databases of traditional Chinese medicine and disease， the network pharmacology was used to screen main active compounds and potential targets of Qinggan Huayu granules for NAFLD and liver cancer. STRING 11.0 was used to analyze the interaction between potential targets. The core targets were selected from the interaction targets by cytoHubba plug-in. The gene ontology （GO） function and the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis were performed on the target by Metascape database. At the same time， <italic>in vitro</italic> experiments were conducted to validate the effect of kaempferol， one of the main active ingredients of Qinggan Huayu granules， on hepatocellular carcinoma cell model and NAFLD cell model. Result:A total of 43 potential targets of Qinggan Huayu granules for for NAFLD and liver cancer were screened， corresponding to 136 active ingredients in 8 herbal medicines. Through enrichment analysis of potential targets， there were 20 biological processes， 13 molecular functions， 9 cellular components and 15 signaling pathways. Qinggan Huayu granules regulated biological behaviors of tumors related to liver cancer and NAFLD （such as apoptosis inhibition and oxidative stress） mainly through kaempferol， quercetin， luteolin and other active ingredients for Caspase-3 （CASP3）， tumor protein p53 （TP53）， vascular endothelial growth factor A （VEGFA） and other hub genes. <italic>In vitro</italic> experiments revealed that kaempferol could inhibit cell proliferation in a dose-dependent manner in hepatocellular carcinoma cell model. And kaempferol could modulate the levels of malondialdehyde （MDA） and glutathione peroxidase （GPx）， which were the molecular markers of oxidative stress of NAFLD cell model. Kaempfero also regulated the expression level of CASP3 in hepatocellular carcinoma cell model and NAFLD cell model. Conclusion:The main mechanism of Qinggan Huayu granules in treating liver cancer and NAFLD with concept of treating different diseases with same method is related to systematic synergy effect of multiple compounds （represented by quercetin， luteolin and kaempferol）， multiple targets （represented by VEGFA， TP53 and CASP3） and multiple signaling pathways （represented by oxidative stress and cell apoptosis）.
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Objective: To explore the potential material basis of Shengmai Injection for the treatment of coronavirus disease 2019 (COVID-19) through network pharmacology and molecular docking technology. Methods : The active compounds of Shengmai Injection were screened by TCMSP and BATMAN-TCM database. The action target was predicted by TCMSP and Targetnet online database, and the active component-action target network diagram was constructed by Cytoscape 3.7.1; Taking "coronavirus pneumonia" as the keyword, coronavirus-related disease targets were searched in GeneCards database and OMIM database. The common target was selected by intersection with the target of Shengmai Injection as the research target. The common target was imported into STRING database to obtain data, and then the protein-protein interaction network map was constructed in Cytoscape 3.7.1 software; The enrichment analysis of GO function and KEGG pathway was carried out by using R language to predict its action mechanism and construct the "component-target-pathway" network diagram; Molecular docking analysis of key targets was carried out by DiscoveryStudio 2.5 software. Results: A total of 22 active compounds were obtained from Shengmai Injection. They were DNOP, β-sitosterol, angeloylgomisin O, gomisin A, gomisin R, wuweizisu C, interiotherin B, changnanic acid, kadsulactone, kadsulignan B, neokadsuranic acid A, neokadsuranic acid B, neokadsuranic acid C, neokadsuranin, schisanlactone A, schisanlactone E, schizandronic acid, uridine, diosgenin, guanosine, N-trans-feruloyltyramine and stigmasterol. There were 224 corresponding targets and 16 common targets with COVID-19, namely CASP3, CASP8, PTGS2, BCL2, BAX, PRKCA, PTGS1, PIK3CG, F10, NOS3, DPP4, NOS2, TLR9, ACE, ICAM1 and PRKCE. The key targets were CASP3, PTGS2, NOS2, NOS3 and ICAM1. GO functional enrichment analysis showed that there were 771 entries for biological processes, 11 entries for cell composition and 79 items for molecular function. A total of 67 signal pathways were screened by KEGG pathway enrichment analysis, which were mainly related to AGE-RAGE signaling pathway in diabetic complications, apoptosis-multiple species, p53 signaling pathway, small cell lung cancer, and so on. The results of molecular docking showed that the components with better docking with the key targets were schisanlactone E, stigmasterol and N-trans-feruloyltyramine. Conclusion: The active compounds in Shengmai Injection, such as schisanlactone E, stigmasterol, N-trans-feruloyltyramine, can act on CASP3, PTGS2, NOS2, NOS3 and other targets to regulate multiple signaling pathways for anti-inflammatory, immune regulation, anti-shock and increasing blood oxygen saturation. This may play a role in the treatment of COVID-19.
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Objective: To predict the targets of the main ingredients in Naozhenning Granule and explore its molecular mechanism of multi-components, multi-targets, and multi-pathways. Methods: Reverse molecular docking (DRAR-CPI) and CooLGeN database were used to predict and screen the targets of Naozhenning Granule; GO enrichment was performed in ClueGO of Cytoscape; KEGG pathway analysis was conducted in DAVID database; The herbs-ingredients-targets-pathways-disease network was constructed in the Cytoscape software. Results: A total of 33 candidate compounds were screened out, and a total of 34 potential targets were revealed for Naozhenning Granule, such as MAPK1, CASP3, and GSK3B. The results of GO enrichment and KEGG pathway analysis indicated that Naozhenning Granule was involved in a series of biological process, such as reactive oxygen species biosynthetic process and positive regulation of smooth muscle cell proliferation as well as some signaling pathways, including PI3K/Akt, MAPK, JAK/STAT, and mTOR. The herbs-ingredients-targets-pathways-disease network suggested that the mechanism of Naozhenning Granule was involved with the regulation of oxidative stress, inhibiting the inflammatory response and the apoptosis of neural cells, regulation of the formation of H2S and the activity of PLG, improving the cognitive function and post traumatic depression. Conclusion: The study suggested that the molecular mechanism of Naozhenning Granule was related with the multi-components, multi-targets, and multi-pathways, which provided a scientific basis for further elucidation of the active ingredients and pharmacological action of Naozhenning Granule.
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Objective:To investigate the anti-proliferation and anti-metastasis effects and study the molecular mechanism of sinomenine in cell line(HepG2).Methods: HepG2 cells were cultured together with different treatment concentrations of sinomen-ine.The effect of sinomenine on inhibition of growth of HepG2 cells were determined by methyl thiazolyl tetrazolium(MTT) assay.The effect of sinomenine on inhibiting metastasis of HepG2 cells were determined by Transwell assay.The inhibitory effect of sinomenine on reverse transcriptase(RT) was studied using inhibitory kinetic method,on the basis,the reactive oxygen species(ROS) of HepG2 cells was monitored by indirect fluorescent labeling.The protein expressions of CASP3,CASP9,CAV1 and SOX2 were analyzed by Western blot experiment.Results: Sinomenine inhibited the proliferation and metastasis of HepG2 cells significantly.Sinomenine had a good inhibitory effect on the growth of HepG2 cells,half inhibitory concentration(IC50) was (15.35±2.43)μmol/L.Sinomenine was RT inhibitor,IC50 was (21.32±2.43)μmol/L.The Western blot showed that CASP3,CASP9 and CAV1 were up-regulated and SOX2 was down-regulated by the sinomenine treatment in HepG2 cells.Conclusion: The potential molecular mechanism of sinomenine suppresses proliferation and metastasis of HepG2 cells by up-regulation of CASP3,CASP9,CAV1 and down-regulation of SOX2.
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This study was aimed to illustrate the interaction mechanism between Chinese herbal medicines and CASP3 target,and to analyze the structural characteristics of CASP3 inhibitors.Molecular docking,molecular dynamics and binding energy were employed to analyze the interactions and mechanism between CASP3 target and ligands which were screened from a series of nature products.The results showed that the binding forces of tanshinone ⅡA and scutellarin with CASP3 target were stronger than others.And the theoretical stable structures of tanshinone ⅡA and scutellarin combined with CASP3 target were obtained by molecular dynamics method.It also can be found that hydrophobic interaction was crucial for tanshinone ⅡA binding to amino acid residues of CASP3 such as Phe256,Ser205 and Trp206.Meanwhile,one hydrogen bond was formed between ligand and receptor.The main interactions between scutellarin and CASP3 target were found to arise from hydrophobic effect in ligand and nine amino acid residues of receptor (such as Ser249,Trp214,and Trp206),four hydrogen bonds with different stabilities and electrostatic interaction.It was concluded that tanshinone ⅡA and scutellarin can form stable structures with CASP3 target.And their similar structures may be useful to screen effective CASP3 inhibitors.