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Aim To explore the anti-inflammatory effect of taurolithocholic acid (TLCA) through network pharmacology-based analyses, to verify with in vitro macrophage study and to reveal the possible mechanisms. Methods The potential targets of TLCA were acquired from public database, and then the protein-protein interaction (PPI) networks against inflammation were constructed and visualized by using Cytoscape. Gene ontology (GO) analyses and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment were performed. The binding activity of TLCA and its target (TGR5) was evaluated through molecular docking analysis. Lastly, the results of the network analysis were confirmed by lipopolysaccharide and interferon-γ induced RAW264.7 cells. Results There were 87 anti-inflammatory potential targets were screened. GO analysis revealed gene functions were mainly involved in regulation of inflammatory response, membrane raft and protein tyrosine kinase. The results of KEGG pathway analysis suggested that PI3K-Akt signaling pathway, human cytomegalovirus infection might be the critical pathways of TLCA against inflammation. The results of in vitro experiments showed that TLCA decreased the LPS and IFN-γ induced inflammatory response in RAW 264.7 macrophages. Furthermore, the expression of TGR5 protein increased after TLCA treatment. Conclusions The potential therapeutic targets of TLCA against inflammation are revealed through network pharmacology analysis. Our results indicate that TLCA might regulate key inflammatory markers through the membrane receptor TGR5.
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Aim To investigate the protective effect of hesperidin (HSD) on the injury of mouse pancreatic beta cells induced by high glucose and fatty acid and the underlying mechanism. Methods MIN6 cells were treated with high glucose and fatty acid after pretreatment of HSD. Cell counting kit-8 (CCK-8) and Hoechst 33258 fluorescence staining were used to determine the proliferation and apoptosis of MIN6 cells. Western blot was used to detect the expressions of apoptosis-related proteins Bcl-2 and Bax. RT-PCR was used to detect the expressions of inflammatory factors tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β). ELISA was used to test the insulin secretion of pancreatic islets. Results High glucose and fatty acid decreased the ratio of Bcl-2/Bax, increased the expression of inflammatory factors TNF-α and IL-1β and inhibited the insulin secretion of mouse pancreatic islets. After pretreatment of HSD, the cell viability and Bcl-2/Bax ratio of MIN6 increased, the expressions of inflammatory factors TNF-α and IL-1β decreased, and the insulin secretion of mouse pancreatic islets increased. Conclusions HSD could resist the apoptosis of mouse pancreatic islet B cell line MIN6 induced by high fat and high glucose, reduce the secretion of inflammatory factors and improve the insulin secretion of pancreatic islets.
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<p><b>OBJECTIVE</b>To investigate the selective inhibitory effect of glycyrrhetinic acid on 4 hepatocellular carcinoma (HCC) cells with different proliferation rates and explore the underlying mechanisms.</p><p><b>METHODS</b>MTT method was used to detect the proliferation rates of 4 HCC cell lines, namely SMMC-7721, SK-HEP1, HEPG2 and HEP3B. Following treatment of the cells with glycyrrhetinic acid (5, 10, 20, 30, 40, and 60 µmol/L), the cell viability was analyzed using MTT assay and the expressions of total ERK protein, p-ERK protein and topoisomerase IIα were detected using Western blotting.</p><p><b>RESULTS</b>Among the 4 cell lines, SMMC-7721 had the lowest and SK-HEP1 had the highest proliferation rate. Treatment with glycyrrhetinic acid for 48 h dose-dependently inhibited the proliferation of all the 4 cell lines in vitro and produced the strongest inhibitory effect in SMMC-7721 cells with the IC of 28.04 µmol/L. The proliferation rate of the cells was positively correlated with the expression levels of p-ERK and topoisomerase IIα, which were the lowest in SMMC-7721 cells and the highest in SK-HEP1 cells. Treatment with 50 µmol/L glycyrrhetinic acid significantly down-regulated the expressions of p-ERK and topoisomerase IIα in the 4 HCC cell lines (P<0.05), while 25 µmol/L glycyrrhetinic acid significantly reduced the expression of topoisomerase IIα and p-ERK in SMMC-7721, HEPG2 and HEP3B cells (P<0.05) but not in SK-HEP1 cells.</p><p><b>CONCLUSION</b>Glycyrrhetinic acid can inhibit the proliferation of different HCC cells particularly in cells with a low proliferation rate. The inhibitory effect of glycyrrhetinic acid might be mediated by reducing the expressions of topoisomerase IIα and inhibiting the ERK pathway.</p>